MXPA99003380A - Compositions and methods for treating viral infections - Google Patents

Abstract

Methods and compositions for treatment, diagnosis, and prevention of a virus comprise administering to a patient antibodies which react with regions of viral proteins and result in neutralization of infectivity and inactivation of functionally essential events in the life cycle of the virus. The antibodies recognize viral epitopes which fail to elicit an immune response in man when encountered through infection or naturally through the environment. In a preferred embodiment, the invention provides compositions and methods useful in the treatment and diagnosis of human immunodeficiency virus (HIV) infections.

Description

COMPOSITIONS AND METHODS TO TREAT VIRAL INFECTIONS TECHNICAL FIELD The present invention relates in general to the treatment and prevention of viral infections. In particular, the invention provides compositions and methods for the production of antibodies and peptides useful in the treatment and diagnosis of human immunodeficiency virus (HIV) infections.

BACKGROUND OF THE INVENTION The diagnosis, treatment and prevention of viral infections is a primary focus of many medical researchers. Although the compositions and methods of diagnosis, treatment and vaccine against various viral infections are known, there are still several viruses which are difficult to detect in man and for which no effective methods of treatment or vaccine are known. Of them, one of the most significant, of course, is HIV. The infectious agent responsible for the acquired immunodeficiency syndrome (AIDS) and its prodromal phases, the AIDS-related complex (CRS) and the lymphadenopathy syndrome (SLA), is a lymphotropic retrovirus called VLA, VLTH-III, VRA and, recently, , HIV, on the recommendation of the International Committee on Taxonomy of Viruses (Ref 299). The nomenclature in the present uses those recommendations for the designated viruses (translated into Spanish) associated with AIDS and its strains. Historical references to strains, which include VLA and VRA-2 are now termed HIV1 LAI and HIV1SR2, respectively. As the spread of HIV reaches pandemic proportions, it becomes a general concern to treat infected individuals and prevent transmission to uninfected individuals at risk of exposure. A variety of therapeutic strategies in different stages of the life cycle of the virus have been sought, and have been pointed out by Mitsuya and Broder, 1987, Nature 325: 773. One approach involves the use of antibodies that bind to the virus and inhibit viral reproduction, either by interfering with the introduction of the virus into the host cells or by some other mechanism. Once the viral component (s) susceptible to antibody intervention are identified, it was expected that sufficient antibody reactivity could be generated to neutralize the infectivity of the virus, and be administered to infected patients. with HIV in the form of immunoglobulins or purified antibodies, and that this passive immunization procedure altered or reversed the progress of HIV infection.

Additionally, it was expected that the vaccine from uninfected individuals with selected epitopes, modified to increase MHC interactions, would provide protection against subsequent infection, after exposure to HIV. It is believed that the envelope glycoproteins of most retroviruses react with receptor molecules present on the surface of susceptible cells, thereby determining the infectivity of the virus for certain hosts. The antibodies that bind to these envelope glycoproteins can block the interaction of the virus with the cell receptors, neutralizing the infectivity of the virus. See, in general, The Molecular Biology of Tumor Viruses, 534 (J. Tooze, ed., 1973); and RNA Tumor Viruses, 226, 236 (R.

Weiss et al., Eds., 1982); González-Scarano and co-authors, 1982, Virology, 120: 42 (La Crosse virus); Matsuno e Inouye, 1983, Jnfect. Immun. , 39: 155 (Virus of neonatal calf diarrhea); and Mathe s and coauthors, 1982, J *.

Immunol. , 129: 2763 (encephalomyelitis virus). Until now, therapeutic strategies aimed at unleashing protective immunological responses in man, by vaccine with HIV proteins / peptides, have failed.

In addition, neither the high-titre neutralizing antibodies, recovered from HIV-infected patients, nor the monoclonal antibodies produced in mice, have been successful in altering the progression from HIV infection to AIDS and death. There is a need in the art to identify alternative immunological targets in HIV, that unleash immune responses that modify the course of HIV infection. The general structure of HIV is that of a ribonucleoprotein nucleus surrounded by a lipid-containing envelope, which acquires the virus during the course of budding from the membrane of the infected host cell. Embedded within the envelope and projecting outward, are the viral glycoproteins encoded. Envelope glycoproteins of HIV are initially synthesized in the infected cell, as a precursor molecule of 150,000-160,000 daltons (gp 160), which are then processed in the cell to an N-terminal fragment of 110,000-120,000 daltons (gp 120) to generate the external glycoprotein; and a C-terminal fragment of 41,000-45,000 dalton (gp41) which is the transmembrane envelope glycoprotein. For the reasons discussed above, HIV gp 120 glycoprotein has been the subject of much research as a potential target to interrupt the life cycle of the virus. Sera from individuals infected with HIV have been shown to neutralize HIV in vitro, and antibodies that bind to gp 120 are present in these sera (Robert-Guroff and co-authors, 1985, Nature 316: 72, Weiss and co-authors, 1985, Nature, 316: 69; and Mathews and co-authors, 1986, Proc. Nati, Acad. Sci. USA, 83: 9709). Purified and recombinant gp 120, stimulated the production of neutralizing serum antibodies when used to immunize animals (Robey and coauthors, 1986, Proc. Nati. Acad. Sci. U. S. A., 83: 7023; Lasky and co-authors, 1986, Science, 233: 209) and a human (Zagury and co-authors, 1986, Nature 326: 249). The binding of the gp 120 molecule to the CD4 receptor has also been demonstrated and monoclonal antibodies that recognize certain CD4 receptor epitopes have been shown to block HIV binding, syncytia formation and infectivity. McDougal and co-authors (1986, Science, 231: 382) and Putney and co-authors (1986, Science, 234: 1392), initiated the neutralization of serum antibodies in animals, after immunizing them with a recombinant fusion protein containing the terminal half. carboxyl of the gp 120 molecule and further demonstrated that the glycosylation of the envelope protein is unnecessary to neutralize the response to the antibody. Shortly after HIV infection, the man's immune system responds to the virus with both antibody production and immune responses mediated by the cells. A review of the immunological responses to retroviruses has been published (Norley, S. and Kurth R., 1994: The Retroviridae, Volume B, J. A. Levy, ed., Pp. 363-464, Plenum Press). Human antibodies, specific for numerous HIV proteins, including gp 160, gp 120, p 66, p 55, gp 41, p 32, p 24 and p 17, have been reported (Carlson, 198, J. Am. Med. Assoc , 206: 674). * The initial antibody response in man to HIV is directed to p7 and p24, followed by gp 120/160, then gp41, p66 / 55 and finally, p32 (Lange, 35 and coauthors , 1986, Br. Med. J., 292-228). As HIV infection progresses to AIDS, antibody levels for P17 and P24 fall markedly to undetectable limits and are replaced by antigens for pl7 and p24. The antibody titers for p32 and p55 also decline, but to a lesser degree (McDougal and co-authors, 1987, "Clin. Invest., 80: 316.) However, substantial amounts of antibodies to gpl60 / 120 persist throughout the course of HIV infection.In the early stages of HIV infection, an elevation in total immunoglobulins, and this increased amount of antibody is specific for HIV and is directed predominantly at gp 120 (Amadori and coauthors, 1988, Clin.Immunol.Immunopathol., 46: 342; Amadori et al., 1989, ". Immunol., 143: 2146). The possible mechanisms for this HIV-specific hyper gamma-globulinemia have been summarized by Bar er E and co-authors, 1995: The Retroviridae, Volume 4, J. A. Levy, ed. Pp. 1-96, Plenum Press. The functional properties and epitopes selected as a target by those antibodies produced during HIV infection have been described and include epitopes that are susceptible to antibody-mediated neutralization. These primary target epitopes are located primarily in the gplβO envelope protein (gpl20 / gp41) and in the gag p! 7 protein; for a summary, see Levy, 1994, Am. Soc. Micro.; Nixon and coauthors, 1992, Im unol. , 76: 515. Neutralizing antibodies to the HIV envelope protein have been identified and bound to conserved and divergent domains at gp 120. These include the regions located at the CD4 binding regions (Linsley and co-authors, 1988 and Thali and co-authors , 1992); the second and third variable curl domains (Fung and co-authors, 1992 and Haigwood and co-authors, 1990) and the carbohydrate portions (Benjouad and co-authors, 1992 and Feizi and Larkin, 1990) . Other neutralization sites have been identified in the outer portion of gp41 and a p7 binding site (Changh and coauthors, 1986). Early studies suggested that the presence of neutralizing antibodies led to a more favorable clinical outcome (Rober-Guroff and co-authors, 1985). However, these studies used selected sera with high neutralizing capacity against HIV laboratory strains and not against autologous HIV isolates (Homsy and co-authors, 1990).; Tremblay and Wainberg, 1990). Subsequent research demonstrated that the autologous antibody had little or no neutralizing activity against autologous HIV isolates (Homsy and coauthors, 1990). The lack of susceptibility to neutralization mediated by the antibody, in the ence of a neutralizing antibody, is believed to be the result of the development of escape mutants that appear after seroconversion (Arendrup and coauthors, 1992) and throughout the infection as new antibody specificities are produced. The clinical relevance of the neutralizing antibodies produced as a consequence of HIV infection is not clear. However, it is clear that, despite a vigorous immune response to HIV in HIV-infected individuals, the progression to AIDS and, ultimately, to death, as a consequence of immunological dysfunction, ominates. Consequently, new methods of treatment are sought.

OBJECTIVES OF THE INVENTION It is an aspect of this invention to identify the viral protein neutralizing regions, which can not trigger immune responses in man, but which do elicit immunological responses in non-human mammals and produce antibodies reactive with those regions. It is another objective of this invention to use those identified neutralizing regions, of the I proteins, and antibodies reactive with them, in the diagnosis, treatment and prevention of diseases caused by the virus. Other objects of this invention will be apparent from the description of the invention detailed below.

BRIEF DESCRIPTION OF THE INVENTION According to the present invention methods and compositions for the treatment, diagnosis and prevention of viral infection are provided by the use of antibodies that react with regions of viral proteins to neutralize and inactivate functionally essential events in the life cycle of the virus. The antibodies recognize viral epitopes that can not trigger an immune response in humans, when they are found during infection or by environmental exposure, but that do provoke an immune response in non-human mammals. Selected epitopes are identified that react with non-human antiviral antibodies, but not with human antiviral antibodies. These epitopes escape surveillance of the human immune system by molecularly mimicking human proteins and, in some cases, are composed of amino acids susceptible to enzymatic decomposition in antigen-processing cells. The desired epitopes are enzymatically divided by human enzymes and, therefore, are not processed by the immunological presentation. Peptides representing these epitopes can be synthesized, optionally modified and conjugated to a macrocarrier adjuvant to elicit antibody responses in non-humans. The preferred adjuvant is a microparticle consisting of multiple repeats of the muramyl dipeptide extracted from Propionibacterium acini. The antibodies and peptides of this invention can be used in immunoassay configurations to identify specific epitope species and to quantitate viral antigens in human tissues and fluids. In a preferred embodiment, the invention provides antibody and peptide compositions and methods, useful in the treatment and. the diagnosis of individuals infected with the virus. In a preferred embodiment, the virus in question is HIV.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides novel compositions and novel methods for diagnosing and neutralizing viral infections. The invention will be described in detail by focusing on a preferred embodiment, in which the virus of interest is HIV. However, it should be understood that the principles of the invention can be used to identify protein neutralizing regions of other viruses and to produce antibodies reactive with those proteins, which can be used to diagnose, treat and prevent infections caused by those other viruses as well. . Emphasizing now on HIV, this invention provides novel compositions and novel methods for neutralizing HIV infections and for preventing or substantially inhibiting HIV infectivity, cell-to-cell transmission and virus production in the infected host. More specifically, HIV protein sequences containing epitopes that can not elicit an immune response in man, when found by infection or naturally by the environment, are used as described in detail below to produce antibodies in non-human mammals. , which can be administered to neutralize HIV infectivity, facilitate the death of infected CD4 lymphocytes and inactivate essential steps in the HIV life cycle. The term "neutralizing region" or "neutralizing region" denotes those portions of HIV, particularly HIV proteins, that contain amino acid segments that define one or more epitopes reactive with antibodies that, either individually or in combination with other antibodies of the present invention. invention, are capable of neutralizing HIV infections.

Appropriate assays to assess neutralization are well known and may include analyzes that measure the reduction of HIV infections in T cell lines, the reduction of VSV pseudotype plaque forming units (HIV) that carry HIV envelope glycoproteins, syncytial inhibition tests and virion-receptor binding tests. The term "inactivating region" indicates those segments of HIV proteins that contain one or more epitopes that, when reacted with antibodies of the present invention, either individually or in combination, inactivate functionally important events in the life cycle. of HIV. Appropriate assays for evaluating the antibody-mediated destruction of lymphocytes infected with HIV are well known and can include antibody-dependent cell-mediated cytotoxicity, complement-mediated lysis and natural killer (NK) analysis. The adequate analyzes to measure the antibody-mediated inactivation of essential steps in the life cycle of the HIV include assays that determine the inactivation of reverse transcriptase or that measure polymerase and protease activity, or that evaluate complement-dependent changes, mediated by antibody, in the permeability of nuclear capsid, which exposes viral RNA to degradation by ribonuclease . When desired, the neutralizing activity can be compared with the reactivity to the antibody in immunochemical assays, such as immunofluorescence, immunostaining, enzyme linked immunoassay and radioimmunoassay. The present invention is based on the discovery that epitopes that are functionally important in the life cycle of HIV, but that are not * immunogenic in man, can be identified and characterized using antibodies produced in selected mammalian species, different from the man. In addition, it has been discovered that the lack of immunological responsiveness in man, to those regions, is a function of the molecular imitation and lack of events associated with MHC, which include the presentation of antigen through events associated with MHC HLA. Class 1 and HLA class 2. With molecular imitation, it is seen that the epitope is autological and does not respond to circumstances below normal. With the lack of events associated with MHC in antigen presentation, several steps are involved and the failure of any step can result in the absence of an immune response to the antigen. Peptide regions containing multiple overlapped epitopes and antibodies to those epitopes have been produced and are shown to neutralize and inactivate essential steps in the life cycle of HIV in vi tro. In addition, patients infected with HIV, who already have AIDS, have been treated with antibodies to these peptide regions, which results in a rapid reduction in the blood-borne infectivity measured by the total arable infectious dose (TCID, acronym by its designation in English: Total Cultureable Infectious Dose). The treatment of chronic AIDS patients with these antibodies has resulted in remarkable clinical improvement, including weight gain, resolution of opportunistic infections, decreased incidence and decreased severity of infections * and fewer medical visits, as well as resolution of the neuropathy associated with HIV. Patients have additionally shown immunological recovery, as defined by a reduction in HIV RNA, increase in the number of CD4, decrease in the number of CD8 and restoration of the cytokine system associated with improved numbers of CD4 and CD8, and their function .

IDENTIFICATION OF EPITOPES AND PRODUCTION OF ANTIBODIES Most immune reactions target immunodominant epitopes. HIV epitopes are very often identified and mapped by various immunological methods that use antisera for HIV, cytotoxic reactivity of T lymphocyte to the HIV epitope targets, and presentation of lymphocyte auxiliary antigen of HIV epitopes. Synthetic peptides of known sequence, which reproduce or mimic the HIV sequence, can be used competitively and non-competitively, using well-known assays to confirm these observations. It should be understood that the stimulation of the immune system can lead either to the increase or to the suppression of the immune response. Factors that govern this include: A. The lymphocyte subpopulation stimulated by the immunogen (suppressor versus helper).

B. The microenvironment, which includes the population of cells residing there, which is in contact first with the immunogen. C. The type of cytokines present in the microenvironment at the moment when the effector cell makes contact with the immunogen. D. The type of cytokines evoked after the effector cell makes contact with the immunogen. E. The structure and biochemical composition of the immunogen. F. The amino acid sequence of the immunogen and its susceptibility to degradation by protease, by the proteases of the microenvironment. From preliminary experiments it was determined that the following properties were fundamentally important for determining the potential value of certain proteins and certain peptides, for use in the production of antibodies that are intended for application in passive immunotherapy, in the treatment or palliation of disease procedures in man and, in particular, HIV infection in all stages, including AIDS: A. The immunogen must lack epitope determinants that are expressed in human cells and human tissues, when they are used to produce the antibody for use in passive immunotherapy, with the following exceptions: 1. The distribution of antigen is restricted to sites sequestered and / or not available for the antibody; 2. The antigen is expressed during the development phases that allow the use of antibody at specific times during the development cycle, when the antigen is not available. 3. The epitope site within the host is not adjacent to a vital structure. 4. The distribution of antigen in the host cell is done at a density lower than that necessary to produce damage, but favorable over the desired target, which results in the direction to the selective destination. 5. Target or target to normal ratio must be sufficiently different and favor the supply of antibody to the desired target or target. B. The number of peptide repeats delivered to an antigen presenting cell directly influences the magnitude of the immune response. C. The epitope must not be present in body fluids at concentrations that neutralize the antibody and prevent direction to the target or target. So far the development of vaccines has focused on designing better technology to amplify responses to the destinations to which the immune system of man responds, and passive immunotherapy has given results that are inconclusive. Described herein are alternative targets or targets in HIV that do not trigger immunological events in man, as well as a new configuration for delivering antigen, which results in immunological reactions to HIV not previously achievable. The methods described herein focus on the treatment of HIV and AIDS, but it should be understood that the formulations of this invention have wide application. The antibody response in goats demonstrates the utility of the invention through the production of antibody to key destinations, in HIV, and as a treatment that results in clinical improvement of AIDS. This technology has wide application in the development of vaccines.

Successful immunological induction to challenge with antigen requires the presentation of multiple epitope repeats by an antigen-presenting cell (APC, acronym for its English designation: Antigen-Presenting Cell) during MHC events. Epitopes that are highly immunogenic have an amphipathic configuration with a hydrophobic amino acid at one end, a hydrophilic amino acid at the other end, containing amino acids consistent with the formation of amphipathic coils, ie, lack spiral-breaking amino acids, such as proline , and lack carbohydrate. Sequences that lack amino acids that are susceptible to protease degradation by proteases present in the microenvironment are especially convenient. To identify targets or immunological targets in HIV with functional importance, immunogenic regions were determined in HIV-related proteins in animal species other than man. Goats were immunized with purified HIV lysate, with and without eliminated carbohydrate groups. The removal of carbohydrate residues from HIV proteins has little effect on the immune response to proteins but may expose hidden epitopes. The commercially obtained HIV lysates were further purified to remove proteins from the origins of the tissue culture, including human HLA class 1 antigens, HLA class 2 antigens and beta-2-microglobulin. After the immunization, goat antisera were tested using competitive immunoassay methodology to identify HIV peptides not recognized by antibodies collected from patients infected with HIV. Deposits of human HIV antisera from sera of selected patients with high neutralizing activity and Western staining were prepared and used as competitive antibodies, using common competitive immunoassay methods. A broad spectrum of goat antibodies was identified that reacted with HIV determinants immunologically distinct from those recognized by human anti-HIV antisera deposits. Those skilled in the art will recognize that other animal species could be used to produce antibodies to those epitopes and that said antibodies could function in reactions mediated by ADCC and its complement. Other animal species suitable for the production of antibodies include, but are not limited to: sheep, rabbits, horses, cattle and mice.

The epitope reactivity of the anti-HIV antibodies was characterized by using 12-mer peptides that cover the linear amino acid sequences of HIVQSF2. Peptides of this size react well with antibodies, can be easily synthesized and can be prepared in highly purified form. The peptides were synthesized by, and acquired from, Purification Systems, Inc. Synthetic peptides were combined with goat anti-HIV antibodies, labeled with peroxidase, and combined with each of two series of microtitre concavities coated with HIV. A series was blocked with anti-HIV from human IgG; the other series, no. The percentage of inhibition of goat anti-HIV that binds to HIV protein sites blocked with human anti-HIV was determined. When inhibition of binding with a specific synthetic peptide was observed, additional peptides were synthesized with amino acid sequences that overlapped those of the original inhibitory peptide, to further define the epitope sequences. The status of the epitopes in the HIV proteins recognized by the goat anti-HIV IgG, but not by the non-human anti-HIV IgG was further evaluated and confirmed using HIV-PRH peptide conjugates as identification markers. In that assay, HIV proteins were absorbed to support said microtiter plate concavities or precision polystyrene beads. 12-mer peptides covering the linear sequence of amino acids from HIV1SF2 to horseradish peroxidase were covalently linked. Human and goat anti-HIV reactivity was measured independently, connecting the anti-HIV reactivity of human and goat with the native epitopes adsorbed to the support and with the peptide epitope covalently bound to the peroxidase. With this procedure, detailed in example 8, only the epitopes contained within the synthetic peptide were recognized. Once the peptides that have significant imitation with human proteins had been identified, the sequences that have functional importance in the life cycle of HIV are determined. This is done, as described below and illustrated in Example 8, by generating antibodies to the candidate peptides and then testing those antibodies for their effect on HIV infectivity and neutralization. As noted above, several specific epitope regions have been identified, and nine are described in detail below, with reference to the HIV1SF2 sequence, unless otherwise indicated. The amino acid residue designations, which are listed below and throughout this application for HIVSF2 are from the Los Alamos data bank (AIDS Virus Sequence Data Base, Los Alamos National Laboratories, Theoretical Division, Los Alamos, NM 87545, E. U. A.). The amino acid residue designations given below and throughout this application, for HIV2NZ • are from the Ex Pasy World Wide Web Molecular Biology server, the Hospital of the University of Geneva and the University of Geneva; and of the BioAccelerator, obtainable through Compugen Ltd, at the Weizman Institute, Israel, and Akira Ohyama, BioScience Systems Department, Mitsuey Knowledge Industry Co., Ltd., Tokyo, Japan. Those skilled in the art will appreciate that additional analog regions ("homologs") of other isolates of HIV, based on their status within related proteins from various isolates. In practice, such homologs can be identified by reference to the HIV1SF2 sequence data, as follows: (a) The amino acid sequences of the HIV and HIV1SF2 isolates can be aligned to obtain maximum homology between the two sequences , generally at least about 75% identification between the sequences; (b) Once an amino acid sequence is aligned with the corresponding site within HIV1SF2 the proteins will show immunological similarity, similarity or identity with HIV1SF2, as defined by the retention of antibody reactivity to the mimicked or homologous sequence. Peptides from other HIV isolates and their amino acid sequences thus identified will typically immunologically mimic the corresponding regions in HIV1SF2. This method to identify key epitopes can be applied to strains of HIV that have not yet been discovered. For example, as new strains of HIV are identified, their envelope and core amino acid sequences can be aligned with that of HIV1SF2 to obtain maximum sequence homology with that strain. The methods by which the sequences are aligned are known to those skilled in the art. When aligning the sequences, it is convenient to maintain the highest possible homology between the cysteine residues. The amino acid sequence (s) of the new HIV strain or species, which corresponds (n) to the location of the peptides specifically described herein, can be synthesized and used according to the invention. It is not necessary for the present invention that the epitopes contained within said sequences be cross-reactive with the antibodies for all strains or species of HIV. Peptides that comprise immunological epitopes that distinguish one species or serogroup from another will be useful for identifying particular species or serogroups and can help identify individuals infected with one or more species or serogroups of HIV. They may also be useful in combination with other peptides, either from a homologous region or from another neutralizing region, in therapeutic regimens. The amino acid sequences of this invention typically comprise about 5 to 50 amino acids and comprise an epitope region or multiple epitope regions, located in HIV proteins, which can not unleash a protective immune response in man, when found by infection or environmental contact, but that do provoke a response in a non-human mammal. Preferably, the sequences comprise between about 5 and 35 amino acids. Synthetic peptides or treated lysates of natural HIV proteins containing the desired amino acid sequences are used to immunize animals that respond immunologically to them, and to produce antibodies having therapeutic value in the treatment of HIV infections. The amino acid sequences or the peptides of interest can not unleash an immune response in man through the imitation of epitopes in human and other proteins. Of particular interest are peptide epitopes shared between HIV proteins and human alpha-fetoprotein, aspartyl-protease, 5'-deoxyuridine triphosphate. Nucleotidohydrolase, cationic eosinophilic protein, eosinophil-derived neurotoxin and ribonuclease precursor 4 and peptide epitope regions, mimicked by neurotoxins of Bungaris naja, Dendoaspis, Psudechis or Androctonus centruroides. In the discussion that follows, reference is made to several human proteins and neurotoxins using normal identification abbreviations for the proteins. A table is given below that indicates those abbreviations and the complete names of the proteins to which they correspond: HUMAN PROTEINS WITH SIMILARITY OF SEQUENCE WITH HIV PROTEINS GSHR Glutathione-reductase / NRM1 Macrofago protein 1 associated with natural resistance TGL3 Protein precursor-glutamine-glutamyl transferase E3 The amino acid sequences of nine of the highly conserved epitope regions discussed above are given below. Three of these regions are in envelope glycoproteins gpl20 (two targets or destinations) and gp41 (one destination); one is in the p66 / 55 reverse transcriptase heterodimer, and one in the plO protease. Other targets or destinations are in the predecessor of Gag (p55 / Gag), with sites in pl7 (two destinations)), p24 and p7. An epitope region in HIV1SF2 gpl20 extends from amino acid residue 4 to 27 and a second region extends from amino acid residue 54 to 76, HIV1. The antibodies for the epitope regions located in gpl20, work synergistically to affect the release of gpl20 from gp41. The release of gpl20 from gp41 is dependent on the dose of antibody and can be demonstrated by neutralization assays, such as TCID, which measures HIV infectivity. An epitope region of a gpl20 neutralizing or inactivating region of HIV2NZ has also been determined. The envelope glycoprotein gpl22 sequence of HIV2NZ has been mapped and approximately from amino acid residue 7 to 43, it is a region that replicates or mimics a sequence of HIV1SF2 gp! 20, and certain human proteins. Targeting the antibody to the region results in the dissociation of HIV2 gpl20 from gp41, which correlates with a reduction in infectivity. A third destination of HIV envelope glycoprotein for HIV1SF2 was localized at amino acid residues 502-541 of the transmembrane glycoprotein gp41. Targeting the antibody from this region in the presence of complement results in antibody-mediated complement-mediated lysis of the envelope glycoprotein of HIV and a marked reduction in HIV infectivity. In addition to the epitope regions of envelope glycoprotein, another epitope region of HIV1, of interest, includes amino acid residues 254 to 295 of the reverse transcriptase heterodimer p66 / 55. Targeting the antibody from this region results in a reduction in reverse transcriptase activity, which depends on the dose of antibody. Also of interest is the epitope region comprising amino acid residues 69-94 of the plO protease. The targeting of antibody from this region results in a reduction in protease activity that depends on the dose of antibody. The reverse transcriptase and protease destinations are in the conserved regions, adjacent to the active enzyme site, which is well known for its mutation and its subsequent resistance to competitive inhibitors. Inactivation mediated by antibody is the result of a steric change or conformation in the enzyme, with secondary loss of activity. This method of inactivation works independently of whether it is not influenced by the mutation in the active site of the enzyme, and is irreversible. Three epitope regions within the Gag gene are also of interest. Specifically, amino acid residues 166 to 181 of the p24 protein of the Gag gene, a target at amino acid residues 1 to 23 and a second target at amino acid residues 89 to 122 of the p7 protein of the Gag gene and residues of amino acids 390 to 410 and 438 to 443 of the p7 protein of the Gag gene are useful in this invention. Antibodies that target such regions result in disruption of the nuclear capsid after lysis of the HIV envelope by the antibodies described above. This direction to destination culminates in the exposure of HIV RNA to RNAse degradation in plasma. Additionally, targets in P17 are exposed on the surface of infected lymphocytes, after budding. This provides an additional soft for lysis with ADCC of the infected lymphocytes. One of the specific peptides mentioned above, comprising at least one epitope not recognized by the antibodies of HIV-infected patients, but recognized by goat anti-HIV antibodies, is the peptide comprising amino acid residues 4 to 27 of the gpl20 envelope protein of HIV1SF2 and its linear epitope-containing subsequences, which has the following sequence: K G T R R N Y Q H L W R W G T L L L G M L M I C.

This peptide mimics the human proteins F0L1, NTCR, PIP5, PPS1, KLTK, MC5R, ECP, INIU, INI9, VPRT, CD69, MYSE, RNKD, ADHE, TC02, LCAT, MAG1, MAG2, MAG3 and LYOX. A second epitope region of the gpl20 envelope glycoprotein of HIV1SF2 extends from amino acid residue 54 to 76, which has the sequence: A S D A R A Y D T E V H N V W A T H A C V P T This peptide reproduces the CYRB and SYV proteins. A third region of epitope, of interest, in the envelope of HIV1SF2 extends from amino acid residue 502 to 541 of the gp41 glycoprotein. This peptide has the following amino acid sequence: HIVl_Env502 R V V Q R E K R A V G I V G A M F L G F L G A A G S T M G A V S L T L T V Q A R 502-541 This peptide reproduces or mimics the human proteins CYPC, TYK2, ACHE, NTCF, NTCR, CD81, 41BL, NIDO, GSHR, C002 and TC02.

In another specific embodiment, an epitope region of interest is that of amino acid residues 2 through 23 of the p7 protein of Gag in HIV1SF2. This peptide has the following sequence: G A R S S V L S G G E L D R W E K I R L R P This peptide reproduces or mimics TFPI, PA2M, BLSA, ECP and FETA proteins and certain neurotoxins, such as NXS1 and NAJAT. The peptide has a hydrophobic sequence that binds to and targets the host cell membrane, and mimics the function of the Sre protein of cell translation. A second target or target in HIV1SF2 p17 extends from amino acid residue 89 to 122. This peptide has the sequence: L Y C V H Q R I D V K D T K E A L E K I E E Q N K S K.

This peptide mimics or reproduces FETA and TRIC. Another peptide of interest is that of amino acid residues 166 to 181 of the p24 protein of the Gag gene and the epitope containing the subsequences. This peptide has the sequence: P E V I P M F S A L S E G A T P This peptide mimics or reproduces the human proteins FETA and TRFL. A third protein epitope region of the Gag gene, of interest, is the peptide having the amino acid residues 390 to 410 and 438 to 443 of the p7 protein of the Gag gene, and the epitope containing its subsequences. This peptide has the sequence: K T V K C F N C G K E G H I A K N C R A P + K I W S S Q.

This peptide mimics or reproduces human FETA and the proteins that bind to RNA. This peptide contains a zinc binding domain that interacts with, and binds to, viral RNA. Antibodies to this region increase the elimination of premature HIV, devoid of envelope, after lysis of infected CD4 + lymphocytes. Also of interest as the epitope region is the peptide of amino acid residues 69 to 94 of the plO protease and the epitope containing its subsequences. This peptide has the sequence: R I G G Q L K E A L L D T G A D D T V L E E M N L P.

This peptide sequence mimics or reproduces the human RENI, BLSA, VPRT and CATD proteins. Antibodies to that sequence inhibit HIV protease activity. Another additional specific sequence, useful in this invention, is a sequence comprising amino acid residues 254 to 295 of the p66 / 55 reverse transcriptase heterodimer of HIV1. This peptide has the sequence: G L K K K K S V T V L D V G D A Y F S V P L D K D F R K Y T A F T I P S I N N E T P.

This peptide sequence mimics the human proteins POLI and ECP. As noted above, other strains of HIV can also be used to obtain peptides and antibodies according to the present invention. Useful peptides from other strains can be determined by comparing and aligning the sequence of another strain with the sequence of HIV1SF2 or HIV2NZ and finding the part of the sequence homologous to the epitopes of interest, identified for HIV1SF2 or HIV2NZ. A sequence of interest in HIV2NZ identified by this method of this invention is in the open reading frame env gpl20 and extends from amino acid residue No. 7 to 43. This peptide has the following sequence: Q L L I A I V L S S Y L I H C K Q F V T V F Y G I P A W R N A S I P L F This peptide mimics the human proteins IL9, SRE1, NRM1, LBP, NOLI, S5A2, LMA1, LECH, LFA3, KPLC, FETA, 3BH2, 3BH1, INR2 and EV2B. For example, once the desired amino acid sequences have been identified, antibodies that recognize those sequences are obtained. These antibodies can be obtained by using proteins containing the peptides isolated from HIV primers, synthetic peptides, bacterial fusion proteins and proteins / peptides from phylogenetically unrelated sources, which contain the desired epitopes.

If viral lysates are to be used, a protein lysate from a single strain of HIV, or a mixture of lysates from two or more different strains, can be used. If a mixture of lysates is used, the mixture may comprise lysates of different strains of HIVl or a combination of at least one cepta HIV1 and at least one strain HIV2.

A preferred mixture is a combination of lysates of HIVIBAL, HIVIMN and HIV2NZ. Viral lysates are initially treated to eliminate lipids and other impurities from HIV proteins. The HIV protein mixture is then treated to remove contaminants of origin in the cell culture, including human leukocyte antigen (HLA), class I and class II antigens. Methods for eliminating those antigens are known in the art and include anti-HLA class I and anti-HLA class II monoclonal antibodies, and immunoaffinity procedures; a method is given in detail in Example 3 below. In addition, it has been found that carbohydrates from HIV proteins must be eliminated; otherwise, phylogenetically preserved carbohydrate determinants would stimulate immune responses when HIV proteins are administered to an animal, which would result in the production of antibodies that would be cytotoxic against human tissues. The proteins are treated with enzymes known to those skilled in the art to remove carbohydrates, including PGNase, neurominidase and glycosidase. One such method is described in detail in the example 3. The mixture of treated HIV proteins can then be used to immunize an animal to produce antibodies to the peptides of interest. Conveniently, the mixture contains approximately equal amounts of the proteins comprising the peptides or the epitope regions of interest. That is, they are conveniently provided in approximate proportions of 1: 1 and the difference in molar ratios between any two peptides is not greater than about 10: 1, preferably 3: 1. Alternatively, the synthetic peptides can be used as an immunogen. If synthetic peptides are used, the amino acid sequence of any desired peptide can be modified, for example, by using a substitute or truncated form of the amino acid sequence.

Amino acid substitutions can be made to avoid the predicted enzymatic cleavage that can occur during the processing of the antigen in a particular portion of the amino acid, to force the amphipathic conformation to satisfy the antigen presentation associated with MHC, and to provide sufficient length for the presentation of HLA a division must occur at or near the epitope boundary. The truncated sequences are selected such that the peptide retains compliance with the epitope length requirements, as predicted by the MHC class 1 and class 2 antigen presentation motifs. More extensive guidelines on substitutions are given below. of amino acid, as part of the section on synthetic peptides. In addition to the substituted and truncated sequences, extended sequences may also be prepared in which additional amino acids are added at either end of a selected epitope region, for the purpose of facilitating attachment to solid phase supports and macromolecular carriers. As an example, the truncated peptide sequences, which extend from amino acid residue 502 to 541 of HIV1SF2 gp41, discussed above, include a peptide with the sequence of amino acid residues 512-531: G I V G A M F L G F L G A A G S T M G A and also a sequence extending from amino acid residue 518 to amino acid residue 527: G A A G Another truncated peptide, particularly useful, is a truncated sequence of the peptide extending from amino acid 7 to 43 of gpl20 of HIV2NZ and having the following sequence: L L I A I V L A S Y L I H C K Q This peptide can be prepared in a variety of ways. The peptide, due to its relatively small size, can be synthesized in solution or on a solid support, according to conventional techniques. Several automatic synthesizers are commercially available today, and can be used in accordance with known protocols. See, for example, Stewart and Young, Solid Phase Paptide Synthesis, 2a. Edition, Pierce Chemical Co., 1984; and Tam and co-authors, J. Am. Chem. Soc., (1983) 105: 6442. Alternatively, you can use technology from Hybrid DNA, when a synthetic gene is prepared using a single filament encoding the polypeptide of substantially complementary filaments thereof; when the individual filaments overlap and can be assembled in a fixing means, in order to hybridize them. The hybridized filaments can then be ligated to form the complete gene and, by selection of the appropriate ends, the gene can be inserted into an expression vector, many of which are currently available without difficulty. See, for example, Maniatis and coauthors, Molecular Cloning, A Laboratory Manual, CSH, Cold Spring Harbor Laboratory, 1982. 0 The region of the viral genome encoding the peptide can be cloned by conventional recombinant DNA techniques, and can be expressed in prokaryotic or eukaryotic expression systems to produce the desired peptides. Preferably the immunogen will be enriched for the desired epitopes to which the antibody-producing B lymphocytes will respond by producing antibodies that neutralize and inactivate essential steps in the life cycle of HIV infection. As used herein, "enriched" means that a desired epitope constitutes at least 25% of the HIV protein, preferably at least 50% and, most preferably, about 95%. More particularly, solutions containing lysate or broken virus extracts, or the supernatant of biologically expressed recombinant proteins, or broken expression vectors or proteins containing imitated epitopes, can be enriched for said proteins, when desired, using methods of purification, such as, for example, polyacrylamide gel electrophoresis. Immunoaffinity purification is a preferred and convenient method for the purification of proteins and peptides containing the desired HIV epitopes, for example, affinity purification using polyclonal or monoclonal antibodies purified by affinity, monospecific. The degree to which the peptides are purified from the solutions for use as an immunogen can vary widely, ie, from about 50%, typically at least 75% to 95%, conveniently from 95% to 99% and, very convenient, to absolute homogeneity.

To obtain antibodies for the desired epitopes, animals are immunized with any of the peptides of interest or HIV proteins containing them, which have been treated to remove HLA carbohydrates and antigens, as described above. The immunization protocols are well known and can vary considerably, but still remain effective. See, Coico, Current Protocols in Immunology, John Wiley and Sons, Inc., 1995. Proteins and / or peptides can be suspended or diluted in appropriate physiological carrier for immunization. Suitable carriers are any non-toxic, biologically compatible substances to deliver and / or increase the immunogenicity of the peptides, including sterile water and 0.9% saline. Alternatively, the peptides can be coupled to a carrier molecule before being used as an immunogen. A preferred technique, for example, discussed in more detail below, involves the binding of the proteins and their fragments to multiple repeats of a glycopeptide, such as the muramyl dipeptide (MDP) to form a microparticle, typically less than 1 miera, and preferably less than 0.2 microns in diameter. The microparticle can then be dispersed in a pharmaceutical carrier for injection. This procedure obtains a high density of the peptide, which can then be used to unleash the desired immune response. The selection of the carrier will vary, depending on the administration route and the response. The compositions can be sterilized by conventional, well-known sterilization techniques. Peptides can be administered by oral or parenteral routes, preferably the latter. The immunogenic amounts of the antigenic preparations enriched for the desired epitopes are injected, generally at concentrations in the range of 1 μg to 20 mg / kg of body weight of the host. The administration can be by injection, for example, intramuscular, peritoneal, subcutaneous, intravenous, etc.

The administration may be once or a plurality of times, usually at intervals of one to four weeks. The immunized animals are monitored for the production of antibody for the desired epitope. The high affinity complement fixative IgG antibody is preferred for passive immunotherapy, and can be used intact, or as fragments, such as Fv, Gab, F (ab ') 2. Antibody fragments may be preferred when greater penetration into tissue is desirable. Antibodies and fragments can be administered alone or as conjugates with toxic substances or isotopes. Once the desired antibody response is obtained, the blood is collected, for example, by venipuncture, cardiac puncture or plasmapheresis. The antibodies are purified from the complex mixture of serum or plasma, according to conventional procedures including, for example, salt precipitation, ion exchange chromatography, size chromatography, affinity chromatography. Frequently a combination of methods is used. Immunoaffinity chromatography is a preferred method. To avoid possible antigenicity in a human receiving the antibody derived from a non-human animal, recombinant antibodies can be constructed. One type of recombinant antibody is a chimeric antibody, in which the antigen-binding fragment, of an immunoglobulin molecule (variable region) is connected by a peptide ligation, to at least part of another protein not recognized as foreign by humans, such as the constant portion of a human immunoglobulin molecule. This can be achieved by fusing exons of variable region of the animal with exons of the constant region, kappa or gamma, human. Various techniques are known to those skilled in the art, such as those described in TCP 86/01533, EP171496 and EP173494, the description of which is incorporated herein by reference. A preferred type of recombinant antibodies is constituted by the antibodies grafted in RDC.

PHARMACEUTICAL FORMULATIONS AND ITS USE The antibodies of this invention that neutralize infectivity kill infected CD4 lymphocytes and inactivate functionally important events in the life cycle of HIV, they are incorporated as components of pharmaceutical compositions. The compositions comprise a therapeutic or prophylactic amount of at least one of the antibodies of this invention, and conveniently a combination of antibodies, with a pharmaceutically acceptable carrier. A pharmaceutically acceptable carrier is any non-toxic, compatible substance suitable for delivery of the antibodies to the patient. Thus, this invention provides compositions for parenteral administration comprising a solution of antibody dissolved in an acceptable carrier, preferably an aqueous carrier. A variety of aqueous carriers can be used, for example, water, water with regulator, saline at 0. 4%, 0.9% saline, 0.3% glycine and the like. These solutions are sterile and are generally free of particulate material. The compositions may further comprise pharmaceutically acceptable auxiliary substances as required for the approximate physiological conditions, such as adjusters and pH regulators, toxicity adjusting agents and the like. For example, sodium acetate, sodium chloride, potassium chloride, calcium chloride, sodium lactate, etc. can be used. The concentration of the antibody in these formulations can vary, typically from less than about 0.1 mg / ml to 150 or 200 mg / ml, preferably between about 1 mg / ml and about 20 mg / ml, and will be selected primarily from based on fluid volumes, viscosities, etc., preferably for the particular mode of administration selected. Determine the concentration of a particular antibody or. of a combination of antibodies, is within the capabilities of one with ordinary experience in the art. A) Yes, a typical pharmaceutical composition for intravenous infusion can be constituted to contain 250 ml of sterile Ringer's solution and 100-200 mg of antibody. Compositions for intramuscular injection can be formed to contain 1 ml of sterile water with regulator and about 20 to 50 mg of antibody. The actual methods for preparing parenterally administrable compositions will be known or apparent to those skilled in the art, and are described in greater detail, for example, in Remington's Pharmaceutical Science, 15a. Edition, Mack Publishing Company, Easton, PA (1980), which is incorporated here by this reference. These compositions may contain a single antibody which, for example, is specific for certain strains of HIV, or for a single protein or glycoprotein expressed by the majority of and, more preferably, by all of the HIV strains. Alternatively, a pharmaceutical composition may contain more than one antibody to form a "combination". For example, a combination containing antibodies against various proteins and strains of HIV would be a universal product with therapeutic or prophylactic activity against the vast majority of HIV clinical isolates. The combination may contain antibodies that bind to epitopes on proteins or HIV envelope glycoproteins, for example, or may contain a combination of antibodies to epitope sites identified above, on the gpl60, gpl20 and gp41 proteins of HIV1SF Env; the p7, pl7 and p24 proteins of Gag; the heterodimer p66 / 55 of reverse transcriptase and protease plO, or a subset thereof; thus neutralizing a series of crucial epitopes in the life cycle of HIV Of course antibodies can also be used for epitope sites in other neutralizing or inactivating regions of HIV proteins. For example, antibodies that modify binding, cell entry, transcription, translation, assembly, target determination of the mature virion for the plasma membrane, and extrusion of the virion will interfere with the events of the cell cycle. life of HIV. Antibody combinations will most often be used to obtain the inactivation of multiple essential HIV proteins. This will be of therapeutic benefit in particular within virions lacking external envelope, but possibly infectious if they manage to enter the cell by other mechanisms, such as micropinocytosis or transfection or the like. The mole ratio of the various antibody components will usually not differ by more than a factor of 10, more usually by no more than a factor of 5, and will usually be at a molar ratio of about 1: 1-3 with each of the other antibody components. With respect to the antibodies for nine specific peptides set forth above, a desirable antibody combination comprises antibodies to the two envelope peptides gpl20 and to the gp41 peptide. More conveniently, the combination comprises antibodies to those three epitope regions plus an antibody to the protease epitope region plO. Even more convenient, the combination comprises antibodies to those four epitope regions plus antibodies to at least one of the other five epitope regions listed. In a highly preferred embodiment, the combination comprises antibodies for the nine epitope regions. The antibodies and antibody combinations of the present invention can be administered independently or given in conjunction with other anti-retroviral agents. The current state of development of other anti-retroviral agents and anti-HIV agents in particular is summarized in Mitsuya and co-authors, Nature, 325: 773-778, 1987. The antibodies and peptides of this invention can be stored in liquid format at various temperatures, which are known to preserve antibody activity, for example, -70 ° C, -40 ° C, -20 ° C and 0-4 ° C, or freeze-dried to store and reconstitute them in a suitable carrier before using them. This technique has been shown to be effective with conventional immunoglobulins, purified antibodies and mixed immunogenic compositions of proteins, glycoproteins and peptides. Freeze-drying and reconstitution techniques known in the art can be employed, and those skilled in the art will appreciate that lyophilization and reconstitution can lead to varying degrees of antibody activity loss (e.g., with conventional immunoglobulins, IgM antibodies tend to have higher activity than IgG antibodies) and that the doses will have to be adjusted to compensate for any loss. The compositions containing the antibodies of the present invention or combinations thereof can be administered for therapeutic and / or prophylactic treatment of HIV infections. In the therapeutic application, the compositions are administered to a patient already infected with HIV, in an amount sufficient to treat, or at least partially stop, the infection and its complications. An adequate amount to achieve this is defined as a "therapeutically effective dose".

The effective amounts for this use will depend on the severity of the infection and the general state of the patient's own immune system; but in general they will vary from 0.1 to 200 mg, approximately, of antibody per kilogram of body weight, preferring doses of 0. 5 to 25 mg per kilogram. The compositions of this invention can be employed in serious disease states that threaten life or in situations that are potentially life threatening. In those cases it is possible, and it can be said that it is convenient for the doctor who is treating to administer substantial excesses of those antibodies. In prophylactic applications, the compositions containing the antibodies herein or their combination are administered to a patient not yet infected with HIV, but who may have recently been exposed to, or is believed to have been exposed to, or at risk of, exposure to the virus (such as, for example, the newborn of an individual infected with HIV) or immediately after exposure or suspected exposure to HIV. If the composition is to be administered to a pregnant woman, infected with HIV, it can be given once or several times before the supply to reduce the infectivity of HIV in the maternal blood and, thus, reduce the risk of HIV transmission to the newborn The newborn at risk can also be treated to further reduce the risk of contracting HIV. An amount defined to be a "prophylactically effective dose" generally ranges from 0.1 mg to mg per kilogram of body weight, depending on the state of health of the patient and the general level of immunity. In addition, the antibodies of the present invention may have use as a carrier molecule, specific to the target or target. An antibody can be bound to a toxin to form an immunotoxin or a radioactive material or a drug to form a radiopharmaceutical or pharmaceutical product. Methods for producing immunotoxins and radiopharmaceuticals are well known (see, for example, Cancer Treatment Reports, 68: 317 (1984)). The heteroaggregates of antibodies of the present invention and human T cell activators, such as monoclonal antibodies to the CD3 antigen or to the gamma Fe receptor on T cells, may allow human T cells or cells carrying Fc-gamma (such as K cells or neutrophils) kill HIV-infected cells by cell-mediated, antibody-dependent cytolysis (ADCC, acronym for its designation in English: Antibody Dependent Cell-mediated Cytolysis). Such heteroaggregates can be assembled, for example, by covalent entanglement of anti-HIV antibodies with anti-CD3 antibodies, using the heterobifunctional reagent 3- (2-pyridyl-dithiol) N-succinimidyl propionate, as described in Karpowsky and co-authors , J *. Exp. Med., 160: 1686 (1984), which is incorporated herein by this reference. Other anti-HIV agents can also be included in the formulations, such as 3 '-azido-3' -deoxythymidine, 2 ', 3'-dideoxycytidine, 2', 3 '-dideoxy-2', 3'-dideshydrocytidine, etc. . In addition to the antibody compositions, compositions comprising the peptides of this invention can be administered for therapeutic and prophylactic vaccine of HIV-infected individuals. For therapeutic application, compositions comprising peptides, either as isolated peptides optionally modified as discussed above, or contained within HIV proteins, treated as described above, and conveniently coupled to a microparticle to further stimulate immunogenicity, are administered to a patient infected with HIV. The amount of peptide administered is selected so as to stimulate the production of antibody to functional epitopes of HIV not previously recognized by the patient's immune system, so that the stimulated antibodies can stop the infection. In prophylactic applications, the compositions of the peptides coupled to the MDP microparticle are administered to persons not infected with HIV to stimulate the production of antibodies against epitopes not otherwise recognized, in order to provide a protective function against future infection.

USES IN DIAGNOSIS AND PROGNOSIS OF THE ANTIBODIES AND THE ANTIGEN The antibodies and epitopes recognized by them and described in the present invention are also useful for the diagnosis and management of HIV infection. Typically, diagnostic assays employing antibodies and / or their respective antigens involve detection of the antigen-antibody complex. Numerous immunoassay configurations have been described and labeled or unlabeled immunochemicals have been employed for that purpose. When not labeled, the antibodies have application, for example, in agglutination analysis, in antibody-mediated complement-mediated cytolysis analysis, and in neutralization analysis. Unlabeled antibodies can be used in combination with other labeled antibodies (second antibodies) that are reactive with the primary antibody, such as antibodies specific for immunoglobulin. Unlabeled antibodies can be used in combination with a labeled antibody, which is reactive with an epitope uncompetitive in the same antigen, such as in sandwich immunoassay, or in combination with a labeled antigen. Alternatively, the antibodies can be directly labeled and used in competitive and non-competitive immunoassays. These types of analyzes and those configurations are well known in the art. A wide variety of labels can be used, such as radioisotopes, fluorescent labels, enzymes, enzymatic substrates, enzymatic cofactors, enzyme inhibitors, ligands (in particular haptens), etc. Numerous types of immunoassays are available and, by way of example, include those described in U.S. Pat. 3,817,827, 3,850,752, 3,901,654, 3,935,074, 3,984,533, 3,996,345, 4,034,074 and 4,098,876. Commonly the antibodies and peptides of the present invention are used in enzymatic immunoassays where, for example, the antibodies of the present, or their respective antigens are conjugated to an enzyme and the immunoanalysis is configured to provide maximum sensitivity and maximum specificity to detect the HIV antigens in biological samples, such as human blood serum, saliva, semen, vaginal secretions or culture suspension of cells with viral infection. It is also possible to design equipment for use with the antibodies herein in the detection of HIV infection or the presence of the HIV antigen. The kits comprise antibodies of the present invention optionally together with additional antibodies, specific for other HIV epitopes. Antibodies, which may be conjugated to a label or label, unconjugated or bound to a solid support, such as the surface of a microtiter plate concavity, or a polystyrene granule, are included in the kits or equipment with regulators, as Tris, phosphate, carbonate, etc .; stabilizers, biocides, inert proteins, for example, bovine serum albumin and the like. In general, those materials will be present in less than about 5% by weight, based on the amount of active antibody and usually present in a total amount of at least about 0.001% by weight, based again on the antibody concentration . It will often be convenient to include an inert extender or excipient to dilute the active ingredients, when the excipient may be present in about 1% to 99% by weight of the total composition. When a second antibody, capable of binding to the antibody, is used, the second antibody will usually be present in a separate ampule. The second antibody is typically conjugated to a label and formulated in a manner analogous to the antibody formulations described above. The epitope of the present, recognized by the antibody, may be provided labeled or unlabeled and may be provided as part of a larger protein (synthetic, recombinant or native), with or without modification, such as the addition of spacer arms, amino groups or cysteine residues, which can be used to fix the peptide to a support and extend it from the surface of the support. These modifications are employed to provide the epitope in an arrangement to raise the immunoreactivity with the antibody to the optimum point. Said peptides are formulated in a manner analogous to that of the epitope-containing proteins, which were described above. The detection of HIV or total virus antigens in various biological samples is useful for diagnosing a current infection with HIV, evaluating the response to therapy, enumerating infected cells, serotype-determining HIV strains, (pods), identifying and quantifying virulence factors associated with primary infection, progression and complications, such as peripheral neuropathy, multifocal leukoencephalopathy and Kaposi's sarcoma. Biological samples may include, but are not limited to: blood, serum, saliva, semen, tissue biopsy samples (brain, skin, lymph nodes, spleen, etc.), cell culture supernatants, eukaryotic expression systems and interrupted bacterial and similar. The presence of viruses, viral antigens, virulence factors and serotype determinants are tested by incubating the antibody with the biological sample under conditions that lead to immune complex formation, followed by detection of complex formation. In one embodiment, complex formation is detected by the use of a second antibody, capable of binding to the primary antibody and typically conjugated to a label. The second antibody is formulated in a manner analogous to that described for the primary antibody formulations described above. In another embodiment, the antibody is fixed to a solid phase support that can then be contacted with a biological sample. After an incubation step, the labeled antibody is added to detect the bound antigen. In another embodiment, the antibody is conjugated to a detection marker and after an incubation step with a biological sample, such as cells or sections of tissue, the sample is evaluated by flow cytometry or microscopy for the antigen.

PREPARATION AND USE OF SYNTHETIC PEPTIDES The peptides of this invention can be modified by introducing amino acid substitutions in the peptide. Substitutions may be convenient to vary one or more particular amino acids, to more effectively mimic the epitopes of different retroviral strains, or to increase the immunological responses or MHC interactions with the epitope, which results in enhanced immunogenicity of the mimicked epitope when it is used for immunization or vaccination. In addition, it may be convenient to make certain amino acid substitutions to increase the chemical stability of the peptide. More specifically, a polypeptide employed in the present invention need not be identical to any particular HIV polypeptide sequence, as long as it is capable of providing immune competence with the proteins of at least one of the HIV strains. Accordingly, the polypeptides of the present may be subjected to various changes, such as insertions, omissions and substitutions, either conservative or non-conservative, where said changes will increase the desired activity of the peptide. Conservative substitutions are substitutions with similar amino acids within a group, such as neutral, acid, basic and hydrophobic amino acids. Examples of substitutions within said groups would include: gly, ala; val, ile, leu; asp, glu; asn, gln; be, thr; lys, arg; phe, tyr; and no, met. Further amino acid substitutions, obtained by applying application programs for molecular models, to the classification of the HLA allotype (DNA and serological) determining data base are shown: In a preferred embodiment of the invention, modifications are made to amino acid so as to replace the hydrophilic residues at the most hydrophilic end of the peptide of interest and the hydrophobic residues at the more hydrophobic end of the peptide. Said substitutions result in the formation of an amphipathic helix with a desired epitope enclosed between the substitutions. The amino acids substituted in the isomer D can be used to enclose epitopes to protect and stabilize the epitope and increase the immunogenicity of the epitope. Since the D-amino acids are not divided by intracellular enzymes, said portion provides peptide epitopes of the desired length for interaction with MHC molecules when they are inserted at the appropriate sites. This is described in detail in example 8.5 below. Usually the modified sequence will not differ by more than about 20% from the sequence of the at least one strain of the human immunodeficiency retrovirus except when more amino acids are added at one or both ends for the purpose of providing an "arm" by means of the which peptide of this invention can be conveniently immobilized on solid phase supports, fixed to macromolecules or modified to increase immunogenicity by altering or increasing the binding of MHC and its presentation. The arms may comprise a single amino acid or up to 50 or more amino acids, and typically are from 1 to 10 amino acids in length. The amino acids, such as tyrosine, cysteine, lysine, glutamine and aspartic acid or the like, can be introduced at the C or N terminus of the peptide or the oligopeptide, to give useful functionality for binding. Particularly preferred is cysteine to facilitate covalent coupling to other peptides or to form polymers by oxidation. Additionally, the peptide or oligopeptide sequences may differ from the natural sequence by modifying the sequence by acylation at the NH2 terminal. (eg, acetylation), amidation with thioglycolic acid or amidation at the carboxy terminus (eg, with ammonia or methylamine) to provide stability, increased hydrophobicity for binding or binding to a support or other molecule, or for polymerization. Thus, for example, in a preferred embodiment of the peptides described herein, one or more cysteine residues or a combination of one or more cysteine residues with spacer amino acids may be added to the peptide ends. Glycine is a particularly preferred spacer when individual peptides are desired. When multiple peptide repeats of the peptide are desired, the peptide of a lysine core is synthesized to form a tetravalent peptide repeat. The configuration is shown as an example. Preferred peptides for use in oxidative polymerization are those in which at least two cysteine residues are added to the ends of a desired peptide. When two cysteine residues are present at the same end of the peptide, there is a preferred embodiment when the cysteine residues are separated by one to three amino acid residues, preferably glycine. The presence of the cysteine residues may allow the formation of peptide dimers and / or increase the hydrophobicity of the resulting peptide, which facilitates the immobilization of the peptide in immobilized or solid phase analysis systems. Of particular interest is the use of the mercapto group of the cysteines or the thioglycolic acids used to acylate the terminal amino groups, or as the first amino acid to constitute repeats of multiple peptides or the like, to bind two of the peptides or oligopeptides or combinations thereof, by a disulfide ligation or a longer ligation, to form polymers containing a number of epitopes. Said polymers have the advantage of increasing the immunological reaction. When different peptides are desired for immunization, they are assembled individually and combined in a combination to give the additional ability to induce antibodies to immunoreact with various antigenic determinants of different HIV isolates. To obtain the formation of antigenic polymers (synthetic multimers), it is possible to use compounds having bis-halogenoacetyl groups, nitroaryl halides or the like, where the reagents are specific for those groups. The binding between the one or two mercapto groups of the peptides or oligopeptides can be a single bond or a linking group and at least 2 or more carbon atoms.

LINKING OF THE MACROMOLECULAR BEARING PEDIATORS The peptide of the present invention can be used to a soluble macromolecular carrier (for example, not less than 5 kDal). Conveniently the carrier can be a poly (amino acid), either occurring naturally or synthetically, to which the antibodies are unlikely to be found in human serum. Examples of those carriers are poly-L-lysine, keyhole limpet hemocyanin, thyroglobulin, albumins, such as bovine serum albumin, tetanus toxoid, etc. The selection of the carrier depends primarily on the final use to which the antigen is destined and on the convenience and availability. In a preferred embodiment, the carrier comprises multiple repeats of glycopeptide to microparticle that can be synthesized or isolated from certain bacteria, such as Propri-bacteribacterium acini or the like. This microparticle is composed of muramyl dipeptide, interlaced extensively, which results in multimeric configurations. When the muramyl dipeptide is isolated from Propionibacterium acini or related organisms, strain selection is useful and selection is based on the chemical analysis of the bacterial cell wall. The preferred embodiment is the muramyl dipeptide extensively interlaced with a dipeptide composed of L-alanine-D-isoglutamine. From preliminary experiments, strain differences have been identified in which the composition of the dipeptide and the length of the peptide vary. Isolates with high concentrations of lipid A and beta-myristate O-acetylated are components of the cell wall.

Preliminary experiments showed that these differences are associated with increases in toxicity and decreases in the adjuvant effect. The selection of strain and the purification of the preferred embodiment are discussed by means of example 4 which follows. The MDP microparticle can be synthesized using procedures known in the art. It has been well established that MDP is a potent immunostimulator, but has significant toxicity. Many attempts to reduce the toxicity of MDP have employed methods to delay release, such as incorporation of MDP into liposomes and other related compounds, or modification of terminal groups. Chemical modification results in a marked reduction in the desired adjuvant effect, and designs have been difficult to control with changing the delivery regime. By way of example, the microparticle configuration of MDP, the size parameters and the methods of fixing the antigen delivery are given below. The separation of the lipids from the microparticle configuration facilitates the rapid internalization of MDP by the cells that present antigen (APC, acronym for its English designation: Antigen Presenting Cells). The predominantly antigen presenting cells are of monocytic lineage and include monocytes, macrophages, histiocytes, Kuffer cells, dendritic cells, Langerhans, etc., and participate in the processing of the antigen and in the presentation of the antigen through events associated with MHC. The factors that contribute to the development of the immune responses to foreign proteins, in part, can be determined by the amino acid sequence and the sequence susceptible to division with protease in the microenvironment. Very often satisfactory immunological responses are observed in peptides forming an antipathetic helix with a hydrophobic end, preferably at the amino terminus and hydrophilic amino acids very frequently at the carboxyl terminal end. Sequence configurations that are resistant to protease degradation and form antipathic helical arrangements are often strong immunogens. The proline-containing residues in the sequence are generally poorly immunogenic, for preventing helix formation and the glycosylation sites are less favorable and, frequently, they inhibit directed responses in the peptide epitopes. The challenge with antigen, which results in a satisfactory immune response in the host animal, requires antigen processing and presentation of the antigen during the events associated with MHC. The exogenous antigen is processed primarily by antigen presenting cells (APC) after internalization in the endosomes. After proteolysis by enzymes, such as cathepsin D, which are present and react in this acidic environment, the peptide fragments that satisfy the criteria described above, are assembled with MHC class II and are presented on the surface of the cell. When the peptides are present in sufficient density, immunological events result. The type of immune response is governed by the density of the peptide by APC, the microenvironment, the cytokine environment and the type of lymphocyte initially stimulated by the antigen presenting cells. After internalization, a cascade of cytokine responses is induced, which modify the microenvironment and establish conditions that lead to immunological events. By way of example, a single microparticle of MDP (0.01 to 0.2 microns) is used to deliver immunogen to antigen presenting cells, which results in immunological responses to poorly immunogenic epitopes not observed using the conventional methods shown in the example 5 that comes later. The quantification of these immunological responses show increases of 10 to 100 times in the concentration of antibody, in comparison with other adjuvants. The peptides herein, used as immunogens, can be linked to the amino acid portion of the carboxyl terminus, of the muramyl dipeptide, using the amino terminus or the carboxyl terminus of the present peptide, or to the aldehyde oxidation product of the carbohydrate, as described in the examples. There will be at least one molecule of the present peptide per microparticle MDP, preferably from 10 to 100 molecules of the peptide of the present per microparticle MDP and, most preferably, 100 to 1,000 peptides of the present per microparticle of MDP. Accordingly, the size of carrier and available linkage groups will influence the number of peptides present per carrier. The macrocarrier composition affects immunogenicity by influencing the preferential uptake of cells, the half-life of the peptide and the presentation of the antigen during immunological events with MHC. One or more other peptides can be linked to the same macrocarrier, but preferably only one peptide is present, either in the monovalent or tetravalent configuration, to the macrocarrier. When immunization with more than one peptide of the present invention is desired, a combination of peptide conjugates of the carrier may be prepared by mixing the individual conjugates at ratios that optimally elevate the immunogenicity of each peptide of the present introduced in the combination. In that configuration, sufficient peptide configuration is available in each macrocarrier conjugate (100-1,000 peptides) to increase antigen presentation by a single antigen presenting cell. The immunogenicity of the peptide of the present will be raised to the optimum by adjusting both the number of the peptides of the present per macromolecular carrier, and the configuration of the presentation, such as amino-carboxyl-binding, the modification of the terminal amino acid and the length and composition of the separating arm, as described. In that configuration the antigen processing by the antigen presenting cell results in a high density, usually more than 100 and, very frequently more than 500 peptides, present on the cell surface of the antigen presenting cell, during the interactions of MHC. With this configuration, significantly higher antibody concentrations occur after immunization, as shown in the examples that follow. The manner of linking is conventional, using reagents such as p-maleimidobenzoic acid, p-methyldithiobenzoic acid, maleic acid anhydride, succinic acid anhydride, glutaraldehyde, etc. The link can be made at the N-terminus, at the C-terminus or at an intermediate site at the ends of the molecule. With multiple repeats of the muramyl dipeptide, the binding of the peptide of the present to aldehyde groups produced by the moderate oxidation of sugar residues, for example, with sodium periodate, after moderate reduction with sodium borohydride and the like, the intermediate becomes to a stable covalent ligature. The number of peptides per microparticle can be controlled by varying the oxidation conditions and quantified using a radioactive tracer. These methods are well known in the art. The fixation method and the configuration of the fixation may vary from one peptide to another, as needed, to achieve the desired response. Various analytical protocols can be used, with which those skilled in the art are familiar, to detect the presence of antibodies to retroviral protein epitopes, or to detect retroviral proteins in complex protein mixtures. Of particular interest is a novel assay described herein, wherein the peptide herein is covalently attached to a detection tag, such as horseradish peroxidase and the native HIV protein expressing the epitope or the epitopes is fixed directly or indirectly to a solid phase support. In that configuration an antibody that recognizes the epitope of the peptide will be bound to the epitope on solid phase, with the epitope on the label. With this method, the reactivity to the epitope of an antibody to HIV can be determined, and it can be quantified by varying the peptides attached to the tag. Peptide epitopes that are associated with the HIV serotype, virulence factors or other characteristics of HIV can be identified and measured in any sample that expresses those epitopes by a competitive one-step immunoassay, described herein, and provided in a manner example.

THE USE OF ANTIBODIES AND THEIR RESPECTIVE EPITOPES IN PURIFICATION PROCEDURES FOR IMMUNOAFINITY The antibodies specific for the epitopes contained within the HIV proteins and the purified proteins containing those epitopes are particularly advantageous for use in the immunoaffinity purification of proteins and peptides containing those epitopes and antibodies reactive with them. In general antibodies will have affinity-association constants of the order of 108 to 1012 M. Those antibodies can be used to purify proteins and peptides containing the epitopes of interest. Genetically modified bacteria can often be used to make the HIV proteins and the recombinant fusion proteins of interest can be purified from the culture medium of the recombinant expression system if the expressed protein is secreted, or from the components of the biological expression system altered, if not secreted, or complex biological mixtures of proteins of which some or a component contains the epitope that mimics an HIV epitope. In general, antibodies that are capable of reacting with HIV epitopes are linked to, or immobilized on, a substrate or support. The solution containing the epitopes is then contacted with the immobilized antibody, under conditions suitable for the formation of immunological complexes between the antibody and the protein containing the epitope. The unbound material is separated from the bound immunological complexes, and the bound proteins of the immobilized antibody are released and recovered in the eluate. Similarly, proteins or peptides containing HIV epitopes or mimicking the HIV epitopes can be fixed to, or immobilized on, a substrate or support, and used to isolate antibodies of interest from a solution. A solution containing the antibodies, such as plasma from which the albumin has been removed, is passed through a column of immobilized peptides or proteins containing the desired epitopes and, after formation of the immunological complex, the antibody is separated. unreactive of the bound immune complex and the antibody is released with an elution buffer and recovered in the eluate. This has particular value for purifying protein containing epitopes that mimic the epitopes of HIV, but derived from phylogenetically unrelated sources with HIV.

Typically, the antibodies are purified in crude from the hyperimmunological serum. Crude ascites fluid or cell culture supernatants and proteins or peptides containing epitopes that mimic HIV epitopes are purified from crude biological sources such as, but not limited to, body fluids, blood, components of blood, cell extracts, tissue extracts of adult and embryonic origin, and culture supernatants, extracts of cultured cells, poisons and recombinant fusion products before attachment to a support. Such procedures are well known to those skilled in the art and can include fractionation with neutral salts at high concentration. Other purification methods, such as ion exchange chromatography, gel filtration chromatography, preparative gel electrophoresis, or affinity chromatography, may also be used to increase the purity of the preparation before use as an immunoabsorbent. The affinity purified antibody can be prepared when desired, by reacting purified crude antibody preparations with a support matrix to which the reactive epitope or epitope-containing protein has been fixed.

Antibodies to HIV epitopes, which mimic them phylogenetically, are of particular interest in nature. Such antibodies are useful as therapeutic agents and are also useful for studying HIV by allowing purification of HIV proteins and mapping of HIV proteins for sequence location and function. These antibodies can be produced by immunization with HIV proteins / peptides, derived from HIV and purified by immunoaffinity by reacting polyclonal, hyperimmune multivalent antisera, with proteins / peptides derived from non-HIV sources, such as embryonic proteins, poisons and microbial / viral components that are not HIV, immobilized in a support as discussed above . The resulting antibody, purified by immunoaffinity, is epitope-specific for one or more epitopes shared by HIV and the phylogenetically unrelated protein used for its immunopurification. These epitope-specific antibodies have particular utility in the immunoaffinity purification of proteins and peptides, both of HIV origin and not originated in HIV. These antibodies can be used to map the location of the epitope in HIV to determine its sequence, assess the functional importance in the HIV out-cycle, its distribution within the HIV sheaths and among other retroviruses, its association with HIV virulence. and, when it is not toxic to man but neutralizing, a crucial function in the life cycle of HIV, is used to treat HIV infection. The support to which the antibodies or epitopes are immobilized, conveniently has the following general characteristics: (a) weak interactions with proteins in general, to minimize non-specific binding; (b) good flow characteristics, which allow it to flow through high molecular weight materials; (c) possession of chemical groups that can be activated or modified to allow chemical bonding of the antibody or epitope; (d) physical and chemical stability under the conditions used to bind to the antibody; and (e) stability to the conditions and constituents of the regulators necessary for the absorption and elution of the antigen. Some commonly used supports are: agarose, derived polystyrenes, polysaccharides, polyacrylamide granules, activated cellulose, glass and the like. There are chemical methods for the binding of antibodies and antigens to substrate supports. See, in general, Cuatrecasas, P., Advances in Eznymology, 36:29 (1972). The antibodies and antigens of the present invention can be fixed directly to the support or, alternatively, by means of a linker or a spacer arm. The general conditions required for the immobilization of the antibody and the antigens to chromatographic supports are well known in the art. See, for example, Tijssen, P., 1985, Practice and Theory of Enzyme Immunoassay, which is incorporated here by reference. The actual coupling procedures will depend slightly on the characteristics and type of antibody or antigen to be coupled. Fixation typically occurs by means of covalent ligatures. An immunological serum, an ascites fluid or a culture supernatant, rich in antibody or an extract or lysate of HIV virus, the supernatant or extract of a cultured biological expression system, the supernatant or extract from an altered cell tissue suspension, or a blood component (adult or embryonic) or other complex mixtures of proteins, such as poisons, body fluids or culture products that contain the epitope. The mixture is incubated under conditions and for sufficient times for antigen-antibody binding to occur, usually at least 30 minutes, more usual, from 2 to 24 hours. The immobilized immune complexes containing the specifically bound antibody or epitopes are then separated from the complex mixture and extensively washed with an absorption buffer to remove unbound contaminants.

The immunological complexes can then be dissociated with an elution buffer compatible with the particular support, the bound protein and the eluate protein.

The eluted protein, the antigen or the antibody is recovered in the eluate. Elution regulators and elution techniques are well known to those skilled in the art. Peptides containing the epitope recognized by the antibody can be used, in the elution buffer to compete for the antibody binding site and elutions can be performed under moderate elution conditions. The protein absorbed selectively from the affinity absorber can be eluted by altering the pH and / or the ionic strength of the regulator or with chaotropic agents. The selection of an elution regulator, its concentration and other elution conditions depend on the characteristics of the antibody-antigen interaction and, once determined, should not be subject to any major change.

The eluted protein may need adjustment to a physiological pH and ionic concentration, if ion concentration regulators or chaotrophic agents are used to dissociate the immune complex. Said adjustment can be made by dialysis or gel filtration chromatography. These methods also allow the eluate protein to regain its natural conformation. The foregoing methods produce, for example, substantially purified proteins which contain epitopes of, or which mimic the epitopes of, HIV and antibodies reactive with epitopes. The purified proteins will typically be more than 50% pure, more usually, at least 75% pure, and often more than 95% to 99% pure. Other aspects and advantages of the present invention will become apparent from the following experimental descriptions, which describe the invention by way of example. The examples further illustrate the process of that invention, but are in no way intended to limit the invention in any way.

EXAMPLE 1 PREPARATION OF HUMAN ANTI-HIV ANTIBODY GROUPINGS (IgG FRACTION), FOR USE IN THE MAPPING OF HIV EPITOPE DIFFERENCES AMONG IMMUNE REACTIONS IN THE MAN AND THE GOATS Human sera from HIV-infected patients were obtained from a community health clinic, with patients' permission; an informed consent was made to each patient, the doctor's approval and approval of the local IRB was obtained. All leathers were originally selected for HIV reactivity at a 1:10 dilution using commercially available test equipment from Abbott Laboratories. Sera with high reactivity (arbitrarily defined by absorbances of the test result greater than 1) was then evaluated by Western blot analysis to identify sera that have antibody reactivities to most HIV proteins (env, gag). and pol.) Patient sera that demonstrated good reactivity to most HIV proteins as defined, were further evaluated by microculture neutralization assays to identify antibody specificities that contain serum that would neutralize HIV infectivity in assays. of microculture (Neutralization of total arable infectious dose (TCID, acronym for its designation in English: Total Culturable Infectious Dose)). We identified and assembled (20-40 ml of each) serum from twenty-nine patients with high antibody titer, reactivity to gpl60, gpl20, p66 / 55, gp41, p24 and pl7 and pLO, and neutralized HIV infectivity in microculture. Further evaluation of the pooled anti-human HIV revealed broad neutralizing activity against multiple strains of HIV and elevated antibody titer (positive by western blotting 1: 100 or more) to the nine epitope regions described in detail above. Human IgG was purified from this pool of sera using conventional procedures. After purification, the total IgG concentration was adjusted to 10 mg / ml, its composition and purity were evaluated by normal immunoassay procedures. The results showed a purity of more than 98% and a composition of human IgG. The purified human anti-HIV was divided into aliquots and frozen for subsequent use in experiments and procedures described below. Those skilled in the art will be familiar with methods for characterizing both antigen and antibody pools to define the specificities that can be used in subsequent determinations against unknown antibodies or unknown antigens, for comparative purposes. As described in the examples that follow, this human anti-HIV was used to track the purification of crude viral ready HIV proteins and map the HIV epitopes recognized by the human immune system and competitive EIA to identify HIV epitopes. to which antibodies produced in goats are destined, but not in man. Western blot analysis was used to characterize the antibody responses in goats immunized with purified HIV proteins containing the peptides of interest, or synthetic versions of those peptides, to evaluate the efficacy of a microparticle carrier complex, designed to amplify immunological responses to poorly immunogenic peptides. Antibodies that neutralized HIV infectivity were evaluated by a microculture procedure that used purified human CDR lymphocytes isolated from peripheral blood mononuclear cells (PBMC), using common techniques and currents that use magnetic particles conjugated to monoclonal antibody to eliminate undesirable cells. The CD4 lymphocytes were stimulated with mitogen to increase their susceptibility to HIV infection and were used everywhere, unless otherwise noted, as the host target target for HIV infection. HIV1SF2 was used everywhere, unless noted otherwise, as the reference HIV strain. The effect of the antibody on HIV infectivity was determined by microculture using techniques familiar to those skilled in the art. Infectivity is expressed as the infectious units (Ul). The reductions in Ul mediated by antibody were associated with neutralization of virus infectivity and expressed as the change in infectious units. The human anti-HIV set was used in all the experiments described here, which require human anti-HIV antibody. This anti-HIV set of human IgG was compared to several commercially obtained anti-HIV human preparations, and had equal or greater HIV neutralizing activity, and when compared by Western blot analysis, they were significantly more reactive.

EXAMPLE 2 CHARACTERIZATION OF VIRAL LISTS OF HIV OBTAINED IN THE TRADE, EMPLOYEES HERE.

Preliminary studies HIVIM, HIV1BAL and HIV2NZ were purchased from Advanced Bio-Technology, Inc., Columbia, MD, U.A., in the form of purified viral lysates. Analysis of these purified viral lysates showed batch-to-batch variation in total protein content, with a scale of 0.8 mg / ml to 1.2 mg / ml. The protein composition of each HIV lysate was evaluated by SDS-PAGE and Western blot analysis, with anti-HIV human IgG. The HIV lysates were treated with protease inhibitors and non-ionic detergents (1.0% volume / volume); as Nonidet P-40 or Igepal CA630, to fully dissociate HIV proteins and glycoproteins to their monomeric form, and clarified by filtration through a 0.22 micron filter. The lipids were eliminated with SeroClear, using common and current procedures. It is well known to those skilled in the art that HIV incorporates human proteins into its envelope as part of the budding process. These contaminants, once identified, were removed by immunoaffinity chromatography. In the initial purification step, the serum contaminants, present in the development medium added to facilitate the growth of the cells, and the contaminants in the HIV lysate, were eliminated by immunoaffinity chromatography, using Sepharose CL6B (2-3 mg of antibody / g of Sepharose) of anti-normal human serum. The chromatography of HIV lysates was carried out in a matrix at a lisate ratio of 1: 1 in volume / volume, at a flow rate of 10 ml / hour. Chromatography and elution were monitored spectrophotometrically at a wavelength of 280 nm. The non-binding fraction, rich in protein, containing the HIV-related proteins, was concentrated at 1 mg protein / ml and stored at -70 ° C for future use, when needed. The bound proteins were eluted by affinity matrices with glycine-HCl buffer, pH 2.2 in 0.9% NaCl. The eluates were neutralized, dialyzed in BPS pH 7.8 containing 0.1% of Igepal CA630, concentrated and stored at -70 ° C for future analysis. Western blotting SDS-PAGE analyzes of the purified HIV preparations consistently demonstrated the presence of gpl60, gpl20, p66 / 55, gp41, plO, p24, pl7 and p7, using human anti-HIV IgG. The HIV proteins were not detected in the glycine HCl eluate, using the Western blot analysis, but the SDS-PAGE gels stained with coomassie brilliant blue for protein visualization, showed 2-3 weakly stained bands.

CHARACTERIZATION OF ANTI-HIV ANTIBODIES PRODUCED TO PARTIALLY PURIFIED HIV LISATES Goats (n = 2) were immunized with the proteins of Purified HIV, obtained before, and responded immunologically with antibodies that reacted with immunodominant epitopes in HIV. Further evaluation of this antiserum showed the presence of cytotoxic antibodies that reacted with lymphocytes infected with HIV and CD4 * uninfected, and red blood cell agglutinins (GR) were detected. These agglutinins were reacted with all blood groups of GR and with GR of rabbits and guinea pigs. Two possibilities for these antibody specificities were considered. One possibility was the contamination of HIV lysates with proteins from the cell culture, and the second was the imitation between HIV proteins / glycoproteins and the glycoproteins found in man and other animal species. It is well known that host membrane proteins are often identified in the envelope of HIV. The incorporation of the host membrane components in the envelope of HIV is thought to be non-specific and is associated with the budding of the mature virion. Two of said proteins previously identified in the mature virion envelope are HLA class I and class II antigens.

Both class I and class II HLA antigens were quantified using an enzyme-linked immunoassay, and the results demonstrated the presence of class I and class II HLA antigens in these HIV preparations, and at concentrations disproportionate to their measured concentration. in cell membrane preparations derived from uninfected culture cells. Other studies confirmed the presence of HLA class I and class II antigens in different preparations (n = 17) of HIV viral lysate. The concentration measured in these preparations was variable but consistently 10 to 100 times higher than that measured in the membrane extracts of infected control cells. The goat anti-HIV antibody was tested by Western blot analysis against known HLA class I and class II isolates and was confirmed to contain antibody specificities directed against HLA class Z, HLA class II (alpha and beta chain) and beta-2 -microglybulin. This antibody was evaluated for its HLA allotype specificity. Commercial trays containing lymphocytes of known allotypes were used as target targets. Under conditions of analysis, this antibody was cytotoxic for all lymphocytes and this cytotoxicity was partially inhibited with class II HLA and soluble class II HLA in a dose-dependent manner (Table 2.1).

TABLE 2.1 INHIBITION BY CLASS I HLA OF LYMPHOCYTOTQXICITY IN ANTIBODY PRODUCED FOR PURIFIED HIV LISTS HLA ciase I and II, soluble, was added to microtiter concavities containing the HIV antibody and incubated overnight at 4 ° C. HLA I and II soluble reduced lymphocytotoxicity, but had no additional effect at concentrations higher than 50 ug. Other studies demonstrated an antibody that reacted with a phylogenetically preserved carbohydrate antigen, present in gpl20, human red blood cells and human white blood cells, and e? red blood cells of different animals, including rabbits, rats and guinea pigs. Absorption studies with human and rabbit red blood cells completely eliminated the remaining antibody activity for both RBC (Table 2.2) and lymphocytes (Table 2.3) after absorption with S-HLA-I and II.

TABLE 2.2 ANTI-V ANTIBODY ANALYSIS ABSORBED, RED ANTI-GLOBULES (GR), FOR RED GLUCOSE AGGLUTININES (GR) CELLS * Antisera produced for purified HIV lysate resulted in GR agglutinins produced in goats after immunization. Goats were immunized (n = 2 of each) with HIVIMN and HIVlBA ?, and HIV2Nz / respectively.

TABLE 2.3 ANALYSIS OF ANTI-HIV ANTIBODIES ABSORBED IN ANTI-GR FOR LYMPHOCYTOTOXICITY DURING IMMUNIZATION * Antisera produced for HIV without treatment with glycosidase at 20 weeks from the date of blood collection and absorbed 0-5 times with red blood cells and tested against lymphocytes for lymphocytotoxicity. After the absorption and elimination of the antibody specificities to HLA and hemagglutinins, the goat anti-HIV antibody was identified by the content of specificities similar to those previously described in the literature. This information demonstrated the need to modify HIV proteins to eliminate carbohydrate and HLA antigens to avoid the generation of cytotoxic antibodies directed against human antigens.

EXAMPLE 3 PURIFICATION AND ELIMINATION OF HALA CLASS I AND CLASS II ANTIGEN AND ELIMINATION OF PROTEIN CARBOHYDRATE HIV SIALDA OF THE HIV SITES HIV lysates were purchased from HIVIMN, HIV1BAL and HIV2N2, from Advanced Biotechnologies, Columbian, MD, E.U.A. They contained protein concentrations on the scale of 0.8 to 1.2 mg protein / ml. Protease inhibitors were added to protect the proteins against degradation and non-ionic detergent (Igepal Ca-630, or Nonidet P-40) was added to dissociate HIV proteins. The mixture was dispensed in capped extraction tubes, with a delipidating reagent to eliminate the lipids and clarify the mixture. The lipids were differentially dissolved in the organic layer, after centrifugation and the aqueous phase was removed. Contaminants of cell culture origin, including HLA, were removed by immunoaffinity chromatography on five separate affinity matrices, prepared by covalent attachment of: immunoaffinity-purified polyclonal IgG antibodies to human serum proteins; monoclonal IgG antibody to HLA-1, monoclonal IgG antibody to HLA-2, monoclonal antibody to beta2 -microglobulin, polyclonal IgG antibodies, purified by immunoaffinity for membrane antigens of lymphocytes and red blood cells.

Each matrix contained 2 to 3 mg of antibody per g of Sepharose CL6B. The columns were configured in a random arrangement and the matrices were poured and equilibrated with PBS at pH 7.8 which contained 0.1% of Igepal Ca-630. The lysate solutions were successively chromatographed in volume equivalent to the volume of the column bed, through each column, at a flow rate of 10 ml / hour. Chromatography and elution were monitored spectrophotometrically at a wavelength of 280 nm. The non-stick, protein-rich fraction containing the HIV-related proteins was concentrated to 1 mg protein / ml. SDS was added to the purified mixture by immunoaffinity, which contained the HIV proteins of interest and the mixture was heated at 70 ° C for 10 minutes. The protein was deglycosylated enzymatically using PGNase. The protein mixture was fractionated by size chromatography on Sepharose G50 previously equilibrated with saline containing 0.1% nonionic detergent. The protein fractions were collected and those containing the desired HIV proteins were collected individually. The three sets of proteins, individually enriched for gpl60 and gpl20, p66 / 55 and gp41, and p24, pl7 and plO, were retained. The fractions were stored at -70 ° C until further processing. Detailed methods for the quantification and elimination of human leukocyte antigen (HLA) are described in Identification, characterization and quanti tation of soluble HLA antigens in circulating and peri tone dialysate of renal patients , F Gelder, Annals of Surgery, volume 213 (1991), incorporated herein by this reference. Table 3.1 shows data obtained from purification.

TABLE 3.1 PURIFICATION OF HIV LISATE The purified HIV preparations as described typically were devoid of contaminants, including HLA class I or class II. SDS-PAGE with Western blot analysis of the purified HIV preparations, consistently demonstrated the presence of gpl60, gpl20, p66 / 55, gp41, plO, p24, pl7 and p7, using the pooled human igG anti-HIV.

EXAMPLE 4 PREPARATION AND CHARACTERIZATION OF REQUIREMENTS BIOCHEMISTRY AND CONFORMATION OF THE MDP MICROPARTICLE TO CAUSE ADJUVANT EFFECTS WITH PURIFIED HIV A multiple repetition of the muramyl dipeptide (MDP), isolated from Propionibacterium acini, formed the central structure of the MDP microcarrier complex of this example. The chemical composition of the monomeric subunit is: CH3CHO-NHCH (CH3) CO-NHCH (CH2CH2-C00H) CONH2 MDP has well-known immunostimulatory properties that have been extensively evaluated in studies designed to determine their effect on the increase of immune function. Those skilled in the art are familiar with this effect. Until now MDP isolates from natural sources and synthetic MDP have been associated with significant toxicity when administered to mammals. That toxicity has limited the effectiveness of MDP as a carrier. A method for the isolation of MDP free of toxic components is provided here. Propionibacterium acini was developed at a medium stationary growth phase and washed to remove contaminants originating in the bacterial culture, employing techniques well known to those skilled in the art. The hydrophobic components contained in the walls of the cells and the cytoplasm were extracted sequentially by successive washes in gradual concentrations of ethanol / methanol / water, at elevated temperatures. The microparticle of MDP resulting in 10% ethanol and its concentration was measured by relating its absorbance at 540 nm with the absorbance of turbidity standards. The concentration of the MDP microparticle was adjusted to 1 mg / ml for storage and subsequent use. Analysis of this preparation showed that the muramyl dipeptide was extensively interlaced with a microparticle size of 0.1 to 0.2 microns. The terminal dipeptide L-alanine-D-isoglutamine linked to amino was identical to the monomeric structure shown above. It is well known that there may be differences between bacterial strains, and that those differences may result in differences in peptide composition, such as terminal peptides with five or more amino acids, changes in the amino acid composition of dipeptide, in particular L-alanine-L-isoglutamine, and sites in which the O-acylated beta-myristate groups had been incorporated. This is not convenient and counts for the toxicity and for the deficient properties of adjuvant of the MDP isolated from natural sources. In a preferred embodiment, the MDP microparticles (0.01-0.2 microns, preferably 0.05-0.1) have the dipeptide L-alanine-D-isoglutamine linked with amino. Said microparticle can be isolated from natural sources, as before, or it can be synthesized using well-known synthesis methods.

EXAMPLE 5 PRELIMINARY PREPARATION AND EVALUATION OF THE MDP- IMMUNOGENIC CONJUGATE The adjuvant effect of the MDP microparticle (0.2 u) of the preceding example on antibody production was evaluated, using a human lambda light chain, monoclonal, poorly immunogenic, lacking approximately 22 amino acids in the sulfhydryl bridge (Mr of 18,000) as immunogen (I). Two conjugates were formed: one in which the immunogen was covalently conjugated to MDP via the terminal carboxyl group and one in which the conjugation was effected by means of the terminal amino group. The MDP-immunogen conjugates were assembled in steps, and the reagent was changed after each step by centrifugation, removal of the supernatant and replacement with the reagent required to continue the conjugation sequentially throughout the MDP: immunogen assembly. The molar ratios are shown with each reaction and the change of reagent that was made after each step prevented the binding in multiple points of the immunogen that, since preliminary experiments, significantly reduced the immunological antibody responses.

SYNTHESIS OF MDP; NH2: IMMUNOGENOUS t CQ2H Protocol for efficient two-step coupling of HIV proteins to muramyl dipeptide, using EDC The following procedure, adapted from a procedure described by Grabarek, Z and Gergely, J., J *. Anal. Biochem, 185: 1311 (1990), allows the sequential coupling of HIV proteins and peptides to MDP without exposing the HIV protein to l-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC) and, thus, affect the carboxyl in HIV. This procedure requires quenching the first reaction with a thiol compound. The reaction is carried out in 2- [N-morpholino] ethanesulfonic acid (MES) (pH 4.5-5.0). 10 mg of MDP, lyophilized in water, was resuspended in 0.5 ml of MES (pH 4.5-5.0) and 0.5 mg (2 mM) of EDC dissolved in MES (pH 4.5-5.0) was combined and reacted for 15 minutes. minutes at room temperature. 2-mercaptoethanol (final concentration of 20 mM) was added to extinguish the EDC and separated by centrifugation. The reaction mixture was washed once with MES and resuspended in 0.5 ml of MES (pH 4.5-5.0). The human lambda light chain fragment, dissolved in MES, was added to the activated MDP at a molar ratio of approximately 2: 1. The pH of the reaction was slowly re-used for a period of 15 minutes at 8.5 by the addition of MES (0.5 M, pH 8.5) and reacted for 2 hours at room temperature. The concentration of lambda light chain fragment added to MDP was calculated from the quantitative analysis of the terminal C02H group of MDP and expressed as moles of C02H per mg of MDP. The reaction was quenched by adding hydroxylamine to a final concentration of 10 mM. This extinction method hydrolyzed any unreacted MDP activation sites and resulted in the regeneration of the original carboxyl. Other means of extinction involve adding 20-50 mM of Tris, lysine, glycine or ethanolamine; however, these compounds containing primary amine will result in MDP of modified carboxyls. When MDP is used with synthetic HIV peptide and modification of that peptide is desired, so as to change the hydrophobicity or bioactive compounds are bound, modification can be achieved by adding the desired compound before the final extinction step. This allows the desired compounds to be added sequentially after initial coupling with HIV peptides. One could also add the bioactive peptides to desired ratios with HIV peptides as a single step, when greater control of the immunogenicity of the peptide is desired. Biological response modifiers, such as IL2, are well known to those skilled in the art and could be used for that purpose. The separation was achieved by centrifugation, a washing step and the suspension again in the selected regulator.

PREPARATION OF MDP-C02H-IMMUNOGENOUS-NH2 Synthesis of MDP; NH2CH2CH2NH2; CQ2H; Immunogen: NH2 Protocol for the efficient three-step coupling of proteins to the muramyl dipeptide using EDC This method allows the sequential coupling of a protein or peptides to MDP without exposing the protein to EDC and, thereby, affecting the amino groups in the protein. The procedure employs two intermediary steps carried out sequentially. The initial reaction is carried out in MES (pH 4.5-5.0). MDP (10 mg) lyophilized in water was combined, suspended again in 0. 5 ml of MES, pH 4.5 and 0.5 ml of EDC (0.5 mg, about 2 mM), dissolved in MES, and reacted for 15 minutes at room temperature. The excess EDC was quenched by adding 2-mercaptoethanol (final concentration of 20 mM) and the activated MDP was separated by centrifugation, washed twice with MES and resuspended in 0.5 ml of MES (pH 4.5). Diaminoethane (NH2CH2CH2NH2) was added, dissolved in MES (pH 4.5), added to the activated MDP at a molar ratio of about 10: 1. The pH was allowed to slowly increase over a period of 15 minutes, by the addition of 0.5 M MES, pH 8.5, and reacted for one hour at room temperature. The MDP: NH2CH2CH2NH2 was separated by centrifugation, washed twice with MES and resuspended in 0.5 ml of MES, pH 4.5. The lambda light chain fragment was suspended in 0.5 ml of MES, pH 4.5 and 0.5 ml of EDC (0.5 mg, about 2 mM) dissolved in MES was added and reacted for 15 minutes at room temperature. The excess EDC was quenched by the addition of 2-mercaptoethanol (final concentration of 20 mM) and the activated protein was removed from the excess reducing agents and the inactivated crosslinkers, by size chromatography, on a gel filtration column, of appropriate size. The activated protein was added to the MDP: activated NH2CH2CH2NH2 at an approximate molar ratio of 5: 1 and reacted for two hours at room temperature. The concentration of protein added to MDP was calculated from the quantitative analysis of the terminal C02H group of MDP, and expressed as the moles of C02H per mg of MDP. The reaction was quenched by adding hydroxylamine to a final concentration of 10 mM. This extinction method hydrolyzed any unreacted MDP activation sites, and resulted in the regeneration of the original carboxyls. If a synthetic HIV peptide is used as an immunogen and modification of that peptide is desired, such as changing the hydrophobicity of the peptide or attacking bioactive compounds, modification can be achieved as described above. Separation is achieved by centrifugation, a washing step and suspension again in the selected regulator.

It should be noted that the bioactive compounds may require the intermediate step described above when fixation is desired by means of the C02H group.

EXAMPLE 6 COMPARATIVE STUDY OF CONJUGATE MDP-IMMUNOGENOUS AGAINST COMMERCIAL ADJUVANTS, WHICH INCLUDE COMPLETE ADJUVANT OF FREUND, RIBI (R), TITER MAX (R) AND ALUM The adjuvant effect of this MDP microparticle (0.1 u) was evaluated by using the human lambda light chain fragment, monoclonal, deficiently immunogenic, described in the preceding example (Mr around 18,000) as immunogen (I). Two conjugates were prepared, one in which the immunogen was covalently conjugated to MDP via the terminal carboxyl group, and one in which conjugation was effected through the terminal amino group. Rabbits were immunized subcutaneously (n = 5 in each group) with about 100 micrograms of lambda light chain, fixed at 500 micrograms of MDP and emulsified in squalene. Animals were immunized at monthly intervals and test bleeds were obtained before immunization and at two week intervals throughout the experiment. The antibody responses to MDP: NH2-I-C02H and MDP: NH2CH2CH2H02C-I-NH2 were comparable in activity; however, rabbits stimulated with MDP: NH2-1-C02H produced at least one additional antibody specificity, determined by competitive EIA.

The antibody responses obtained were compared with those obtained when conventional adjuvants were used for antibody response, including complete Freund's adjuvant, Ribi (R), Titer Max (R) and Alum (aluminum hydroxide). Both MDP: H02C-I-NH2 and MDP: NH2-I-C02H were significantly superior to conventional adjuvants with immunogen, by inducing the antibody. The immunogen concentration (100 ug / immunization) and the immunization pattern were identical in all groups. Table 6.1 shows the antibody titer measured at biweekly intervals and the titer is expressed as the reciprocal of the dilution that produces a positive reaction as described above. Both MDP conjugates were superior to conventional and well known adjuvants.

TABLE 6.1 T-0 = Blood collection before primary immunization and pre-immunization. MDP: I = muramyl dipeptide: immunogenic microparticle (< 0. 2 m). * The peptide in the terminal amino group was conjugated to isoglutamine with the exposed carboxy terminus (MDP: I: C02H).

+ The peptide was conjugated at the carboxy terminus, by means of an intermediate step, using diaminoethylene to modify the carboxyl terminus of MDP. (MDP: I: NH2).

EXAMPLE 7 RESPONSE TO MONONUCLEAR CELL CYCLOIN OF PERIPHERAL BLOOD, INDUCED WITH IMMUNOGENS, MD2; NH2; I; CQ2H and MDP; NH2CH2CH2NH2: Q2HC-I-NH2 To further evaluate the mechanisms associated with the increased antibody response to the MDP-immunogen microparticle complexes, an in vitro method that measured the production of cytokine or peripheral blood mononuclear cells was used and compared with known cytokine inducers. Lipopolysaccharide (LPS) and LPS were used with phytohemagglutinin (PHA) as known cytokine inducers. The cytokines were quantified using a well established analysis and expressed as units / ml. Peripheral blood mononuclear cells were isolated by gradient centrifugation of Ficol Hypague and adjusted to a concentration of 2 x 106 / ml in tissue culture medium. Plaques were formed with cells (100 ul) in microculture concavities. MDP was added: I: C02H, MDP: I: NH2, PHA + LPS, LPS and medium alone, undiluted or diluted to 1:10 and 1:25 (10 ul.) The cultures were incubated at 37 ° C in a 5% C02 atmosphere for 48 hours, and the supernatants were removed and analyzed by normal bioassays and / or EIA methods.

TABLE 7.1 Both MDP: I: C02H and MDP: I: NH stimulated greater type I cytokine responses than LPS + PHA or LPS alone. The cytokine type 1 responses increased the immunological events, while the cytokine type 2 response is indicated by high levels of gamma IFN and IL2, and lower levels of TNF and IL6.

EXAMPLE 8 CHARACTERIZATION OF THE ANTIBODY RESPONSE IN GOATS, A PROTEINS OF HIV NOT TREATED AND TREATED TO ELIMINATE THE PORTIONS CARBOHYDRATE AND COMPARISON OF PROPERTIES ADJUVANTS OF MDP MICROPARTICLES WITH ADJUVANTS CONVENTIONAL Example 8.1 Viral lysates HIVIMJ, HIV1BAL HIV2N2 FROM Advanced Biotechnologies Inc., Columbia, MD, E.U.A. were purchased, and half of each preparation was treated enzymatically to remove carbohydrate and all preparations (with or without carbohydrate) were purified as before.

An aliquot of each was conjugated to MDP by means of the amino terminal residue, according to the procedures indicated in example 5, and was individually suspended in squalene. For comparison, these HIV proteins were also emulsified in complete Freund's adjuvant, without conjugation to MDP.

Group 1 - HIVIMJIVIHIBA: 1 without elimination of carbohydrate. Group 2 - HIVIMUIVIHIBA / 1 1 with elimination of carbohydrate. Group 3 - HIV2NZ without elimination of the carbohydrate. Group 4 - HIV2NZ with carbohydrate elimination Group 5 - HIV1MN: HIV1BA: HIV2NZ, 1: 1: 1, without elimination of carbohydrate. Group 6 - HIV1MN: HIV1BAL: HIV2NZ, 1: 1: 1, with elimination of carbohydrate. Group 7 - HIV1MN: HIV1BAL: HIV2NZ, 1: 1: 1, without elimination of the carbohydrate and emulsified in complete Freund's adjuvant, without conjugation to MDP. Group 8 - HIV1MN: HIV1BAL: HIV2NZ, 1: 1: 1, with elimination of the carbohydrate and emulsified in complete Freund's adjuvant, with conjugation to MDP. The goats were stratified in immunization groups 1-8 (n = 3 each) and immunized respectively at intervals shown in Table 8.1 with 100 ug of HIV / immunization. Blood samples were obtained before immunization and at biweekly intervals. The antibody reactivity was quantified by EIA using the commercially available analysis equipment, approved by the FDA, of Abbott Laboratories.

The results were expressed as the reciprocal of the dilution of antisera that produced an absorbance value > 1.0.

TABLE 8.1 ANALYSIS OF ANTI-HIV ANTIBODY RESPONSE TO HIV PROTEINS TABLE 8.1 (continued) T-0 = Blood collection of primary immunization and pre-mmumzacion. * - immunization and reinforcer. Antibody responses were measured at biweekly intervals throughout the immunization. There was no significant difference in antibody reactivity, as measured by EIA, between individual groups of animals (groups I, 2, 3, 4, 5, and 6) with MDP. The removal of the carbohydrate had no effect on antibody production for the desired anti-HIV proteins.

EXAMPLE 8.2 The antisera obtained was evaluated as described in example 8.1, for the hemagglutination of antibodies. As can be seen from Table 8.2, the removal of carbohydrate from HIV proteins obliterated the haemagglutination response.

TABLE 8.2 ANTI-HIV ANTIBODY RESPONSE ANALYSIS FOR RED BLOOD CELLS TABLE 8.2 (continued) T-0 = Blood collection of primary immunization and pre-immunization. 1 - Carbohydrates removed * Immunization and booster HIV preparations treated to remove carbohydrate groups could not produce antibody reactivities capable of binding red blood cells (Table 8.2A). Goats immunized with MDP-HIV conjugates without elimination of carbohydrates and a goat immunized with purified HIV, without elimination of carbohydrate, emulsified in complete Freund's adjuvant, produced red blood cell agglutinins. There was no detectable difference in the titer of red blood cell agglutinins in antisera from goats immunized with MDP-HIV conjugates or with HIV emulsified in complete Freund's adjuvant. The red blood cell agglutinins described herein were essentially identical to those described in the preliminary studies of Example 2. These agglutinins were cytotoxic (Table 8.2A). However, there was no detectable antibody reactivity to HLA class I or class II and absorption with red blood cells completely eliminated hemagglutination and cytotoxic antibody reactivity.

TABLE 8.2A ANTI-LISIS SEQUENCES OF ANTIBODIES ANT -V H FOR LYMPHOCYTOTOXICITY TABLE 8.2A (continued) T-0 = Primary immunization and blood collection prior to immunization. 1 - Carbohydrates eliminated. * Immunization and Booster To assess the possibility of imitation between the HIV and RBC carbohydrate groups, additional antisera were produced by immunizing a goat with an immunogen composed of purified and pooled cell membranes isolated from human red blood cells. Western blot analysis employing these antisera demonstrated reactivity with a red blood cell glycoprotein (Mr about 35,000) and reacted with gp41 and HIV gpl20. However, gp41 and HIV gpl20 treated to remove the carbohydrate groups were not reactive. These catos were consistent with the phylogenetic imitation between the carbohydrate epitope in HIV and the red blood cell glycoproteins. Western blot analysis demonstrated strong reactivity to most HIV proteins, including gpl60, gpl20, gp41, p66 / 55, plO, p24, pl7 and p7. There was no apparent difference in the reactivity or specificity of these antibodies for the HIV epitopes described herein. Those antibodies reacted for all HIV isolates tested, including those that had been shown to have resistance to reverse transcriptase and protease inhibitors.

EXAMPLE 8.3 NEUTRALIZATION OF HIV INFECTIVITY BY ANTIBODIES PRODUCED FOR HIV WITH DELETED CARBOHYDRATE This example describes and characterizes the neutralization of HIV infectivity using the antibodies produced for HIV with eliminated carbohydrate, described above. The results indicate that these antibodies contain high levels of neutralizing activity and protect the CEM cells against infection in a manner that depends on the dose.

Neutralization analysis A sensitive neutralization analysis was used to quantify the effect of goat anti-HIV on HIV infectivity. The C4 CD4 cell line, which is highly susceptible to HIV infection, was selected as the target cell to determine the effect of this antibody on HIV infectivity.The antibody was made and dilutions as required in RPMl medium. containing 10% fetal calf serum, a suspension of HIV1SF2 was harvested from cultures of about 4 days of CEM in the logarithmic development phase, filtered through 0.2 or 0.45 micron filters, aliquoted and frozen -70 ° C. An aliquot was thawed, titrated to determine the ITC50, and subsequent analyzes were performed with freshly thawed aliquots, diluted 1: 500 in culture medium, at a concentration approximately ten times the amount required to infect the 50 % of the CEM cells in the culture (10 times the DITC50) The virus suspension was mixed with an equal volume (250 ul) of dilutions to the 5-fold antibody from 1: 5 to 1: 9,765,625. The virus / antibody mixture was incubated for 60 minutes at 37 ° C and duplicate samples of 200 ul were used to inoculate concavities containing 1.0 ml of about 2xl05 CEM cells per concavity. The cultures were incubated at 37 ° C in a humidified atmosphere, with 5% C02 for 14 days. Cells were harvested, pellets formed and lysed with 1% Triton X-100 in PBS for about 10 minutes. The amount of virus (or viral antigen) present in the cells subjected to lysis was quantified, using a p24 analysis commercially available. The titer of the neutralizing activity was determined as the reciprocal of the antibody dilution that inhibited the production of the p24 antigen in more than 50% goat pre-immune IgG, prepared in a similar manner. 200 microliters of the cell suspension subjected to lysis was analyzed.

TABLE 8.3A ANALYSIS OF NEUTRALIZING ACTIVITY OF ANTI-HIV ANTIBODY TABLE 8.3A (continued) T-0: Primary immunization and blood collection before immunization 1 = eliminated carbohydrates. * immunization and reinforcement ** Anti-HIV preparations obtained from the 20 weekly bleeds were absorbed with human red blood cells and both the unabsorbed and absorbed samples were tested under identical conditions. The results are expressed as neutralization title (not absorbed / absorbed). Anti-HIV preparations were produced with the highest neutralizing activity for HIV-MDP conjugates that were not treated to eliminate the carbohydrate determinants (Table 8.3A). Absorption in red blood cells of antibodies to HIV conjugates with eliminated carbohydrate had no effect. However, absorption in red blood cells of antibodies to HIV conjugates with the intact carbohydrate resulted in a significant reduction in neutralizing reactivity, confirming the presence of phylogenetically present carbohydrate moieties shared between HIV and humans (bottom row , box 8.3A). All anti-HIV antibody preparations produced for HIV-MDP conjugates were statistically higher than anti-HIV produced using Freund's complete adjuvant. Due to the predicted genetic variability characteristic of HIV, anti-HIV of the twenty weekly preparations for neutralizing activity was tested using HIVMN and four wild-type isolates of HIV, including one characterized as a multi-drug resistant strain, under conditions identical to those described above. The anti-HIV of the 20 weekly antibody preparations produced for HIV conjugates devoid of carbohydrate, neutralized all the strains (Table 8.3B). Those skilled in the art will recognize that the neutralizing activity produced for deleted HIV on HLA proteins, with eliminated carbohydrate, is considerably greater than the neutralizing activity typically observed in human anti-HIV sera.

TABLE 8.3B ANALYSIS OF ANTIBODY NEUTRALIZING ACTIVITY ANTI-HIV AGAINST MULTIPLE CEPAS 1 = eliminated carbohydrates EXAMPLE 8.4 EFFECT OF ANTI-HIV ON HIV-INFECTED CD4 LYMPHOCYTES IN COMPOUND MEDIATED CITOTOXICITY, DEPENDENT ON ANTIBODY Antibodies against HIV conjugates with eliminated carbohydrate and with intact carbohydrate were evaluated for the reactivity of complement-mediated cytotoxicity to normal peripheral blood mononuclear cells, enriched for CD4 lymphocytes with and without HIV infection. Peripheral, normal mononuclear cells were isolated; they were subjected to centrifugation with Ficol Hypaque gradient and enriched for CD4 'lymphocytes. The CD4 'lymphocytes were stimulated with PHA and infected with HIVMN for 7 days in microculture. The supernatants were removed and replaced with anti-HIV, produced for HIV preparations after elimination of the sugar group, and showed no cytotoxic effects on normal cells. As can be seen from Table 8.4, the anti-HIV lysis of infected CD4 lymphocytes, in a manner that depended on the dose.

TABLE 8.4 MEDIATED CYTOTOXICITY ANALYSIS BY ANTI-HIV ANTIBODY OF INFECTED CD4 LYMPHOCYTES TABLE 8.4 (continued) T-0 = primary immunization and blood collection prior to immunization. 1 = eliminated carbohydrates * immunization and reinforcement.

EXAMPLE 8.5 SYNTHESIS OF PEPTIDES THAT CORRESPOND TO THE SEQUENCE OF AMINO ACIDS OF HIV PROTEINS Synthetic peptides were constructed as dodecane peptides that mimic or reproduce the amino acid sequence of HIV1SF2. Amino acid sequences for gp 120, gp 41 Vif, gag p 17, gag p 24, nef, Rev, integrase, protease, Tat: HxB2 and reverse transcriptase and reverse transcriptase with overlaps by six amino acid residues, were synthesized by and purchased from Purification Systems Inc., using solid phase technology. Butyloxycarbonyl-S-4-methylbenzyl-L-cystine coupled with polystyrene was used, using dicyclohexylcarbodiimide with a catalytic amount of 4-N, N-dimethylaminopyridine as a solid phase support for the synthesis. The amino groups were protected with terbutyloxycarbonyl (t-BOC) and the side chain protecting groups were as follows: benzyl ether for the serine hydroxyl, dichlorobenzyl ether for the phenolic hydroxyl tyrosine, and the beta-benzyl esters were used for the carboxyl groups in glutamic acid and aspartic acid, respectively. Trifluoroacetic acid (40% in methylene chloride) was used to remove t-BOC and the resulting salt was neutralized with N-diisopropylethylamine (10% in methylene chloride). Diisopropylcarbodiimide was used to couple t-BOC-amino acids. The protecting groups were removed and the peptide was cleaved from the resin at 0 ° C with anhydrous hydrogen fluoride containing 10% anisole and 1% ethanedithiol as scavengers. The hydrogen fluoride reagent was removed under vacuum at 0 ° C and then the peptide was precipitated and washed with anhydrous ether. After extraction of the peptide from the resin, with trifluoroacetic acid, the solvent was evaporated at 15 ° C and the peptide was precipitated again with ether. The ether was decanted after centrifugation and the pellet was dissolved in 5% acetic acid with 6M guanidine hydrochloride.

This solution was desalted on a BioGel P2 column equilibrated in 5% acetic acid and the fractions containing peptide were combined and lyophilized. A cysteine residue was added to the carboxyl terminus of the peptide, as necessary, to give a functional SH group for coupling the peptide to carrier proteins or to a solid support for EIA or MDP methods (example 5). When multiple repeats of the peptide were desired, synthesis was conducted by first attaching a cysteine residue to the resin support. Carbon separators of various lengths were added; the selection of separator length varied and depended on the application, of the peptide loading and of the length and spherical influences predicted from preliminary data resulting from peptide fixations to the supports. A six-carbon separator was first fixed, such as 6-aminohexanoic acid with additions of lysine (lysine) -2- (lysine) 4 as described above, with diaminoethane in example 5, but altering the blocking sequence of the protecting group. The amino groups were protected and then deprotected to allow two lysine residues to bind to the unprotected amino terminus, deprotection, followed by addition of lysine, formed a branched chain structure for peptide synthesis. Peptides with specific biological function or with sequences that are susceptible to enzymatic degradation were modified by the addition of D-amino acids. A particularly useful addition is the addition of L-alanine-D-isoglutamine with the peptide of interest synthesized from the NH2-terminus of D-isoglutamine. In another arrangement, the peptide was synthesized with L-lysine-L-lysine-peptide-D-isoglutamine. The carboxy-terminal lysine groups are highly susceptible to enzymatic degradation by many enzymes in the microenvironment, while D-isoglutamine results in an increase in the half-life of the peptide and provides a hydrophobic site for assembling peptides that require anfaphatic properties to elicit a function such as receptor binding and immunological induction through events associated with MHC. A tyrosine residue was added to the amino terminus for radioactive labeling with 12Syode to determine the coupling efficiency of peptide to carrier protein, and to identify the peptide during purification. 125I also provided a tracer to track the half-life of the peptide in biological systems and evaluate receptor binding when the function of the peptide was not affected by the addition of tyrosine.

EXAMPLE 8.6 USE OF SYNTHETIC PEPTIDES THAT IMITATE HIV PEPTIDE SEQUENCES AND OTHER RETROVIRUS FOR IDENTIFICATION OF EPITOPES VIRAL Within this invention, synthetic peptide sequences that mimic the highly conserved sequences found in HIV and other retroviruses are disclosed and their functional significance as immunological targets in the treatment of viral infections is identified. These peptides have additional application in the diagnosis and management of HIV infection that is the result of HIV microvariants with sequences that contribute to the pathogenesis of HIV through non-specific down regulation of immunological reactions, induction of autoimmunity and through effects toxins that lead to peripheral neuropathy associated with HIV.

Each of these events, when presented in a patient, contributes to the pathogenesis of HIV and declines the quality of life. The synthetic peptides used to identify and quantify those regions of HIV peptide that initiate these events are useful for identifying risk factors for autoimmunity and peripheral neuropathy. The synthetic peptides provide additional utility in a novel screening procedure to monitor the progression of the disease and changes in the advance as a result of the treatment. In one step of this invention an enzyme immunoassay (EIA) was configured for the identification of goat antibody specificities on HIV not recognized by human anti-HIV. Purified preparations of the HIVpl gpl20 and gp41 proteins were applied to concavities of polyvinyl microtiter plates at 5 ug / ml in phosphate buffered saline (PBS <pH 7.8) by incubation for 20 hours at 37 ° C. The concavities were washed with PBS containing 0.1% Tween 20 (PBS / Tween) and the unoccupied sites of each concavity were saturated with 5% bovine serum albumin by incubation for one hour at 37 ° C. Plates were used immediately or stored at 4 ° C. 100 ul of human IgG anti-HIV (example 1) was added to each concavity, incubated for 25 hours at 4 ° C and the concavities were washed with PBS / Tween. Goat anti-HIV, previously produced and labeled with HRP, was added by common procedures, to the concavities, at a dilution necessary to produce an absorbance of 1.0 under the conditions of the analysis (1: 10,000 titer) and incubated for 24 hours. hours at 4 ° C. The concavities were washed and substrate was added to determine the amount of goat IgG binding. The percentage of inhibition of the binding induced by blocking HIV proteins was calculated with human anti-HIV, using the formula: (DO without blockade - DO blocked) x 100 OD of negative control blocked DO without blockade The minimum blockade of goat anti-HIV-HRP conjugate for human HIV antigen was indicative of the goat anti-HIV binding different from the anti-HIV one. HIV human. In another step, the EIA was configured to identify the epitopes on HIV proteins that were antigenic with goat antibodies, but not with human antibodies. Purified preparations of HIVpl gpl20 and gp41 proteins were applied on 5 ug / ml polystyrene granules in PBS by incubation for 24 hours at 37 ° C. The granules were washed with PBS containing 0.1% Tween 20 (PBS / Tween) and the unoccupied sites in each granule were saturated with 5% bovine serum albumin, incubating for 1 hour at 37 ° C. The granules were used immediately or stored at 4 ° C. 100 ul of anti-HIV human IgG (example 1) was added to each granule, incubated for 24 hours at 4 ° C. 100 ul of human IgG anti-HIV was added (Example 1) to each granule, incubated for 24 hours at 4 ° C and washed the granules with PBS / Tween. Synthetic peptides were dissolved in PBS containing 5 mg / ml bovine serum albumin and 0.1% Tween 20, at a concentration of 0.1 mg / ml. 25 ul of peptides added to 100 ul solution of the goat anti-HIV conjugate-IgG-HRP and incubated for 24 hours at 4 ° C. The mixture was then added to two series of granules coated with HIV. One series was blocked with human anti-HIV that was added to two series of granules coated with HIV. A series was blocked with human anti-HIV that was added at the same time as the peptides were added to the goat anti-HIV-HRP conjugate. The granules with the reagents were incubated for 24 hours at 4 ° C. After the incubation the granules were washed and the peroxidase activity was measured, as described before. Activity was plotted against the position of the peptide within the HIV proteins. These graphs showed areas of the HIV proteins used as targets by goat HIV immune IgG, which was not recognized by the human antibody. When inhibition of binding with a specific synthetic peptide was observed, more peptides were synthesized to overlap the original peptide by peptides of additional lengths. The lack of inhibition by the synthetic peptides was considered to represent the lack of immunological target by the goat immune system. Using this procedure, linear peptide epitopes were selected for study. In another preferred embodiment of this invention, an EIA was configured to use peptide-peroxidase conjugates, produced as described below, to identify epitopes reactive to the antibody in HIV proteins, recognized by the goat immune system, but not by the human . The peptides that mimic the HIV sequences covalently linked to HRP were covalently linked to produce a library of enzyme-labeled peptides for use in mapping the antibody specificity. Specifically, HRP was supplied to a reaction tube, at a calculated molar ratio of 1 part HRP: 10 parts of peptide. Each peptide was supplied individually to be coupled, to microtubes at a concentration of 0.1-1 umol / ml, at volumes of 0.1-1 ml, each of which varied, depending on the amount of conjugate desired and the molecular weight for the peptide. HRP was dissolved at a concentration of 1-10 umol / ml in 0.1 M carbonate / bicarbonate buffer (pH 9.8) at 4 ° C and sodium periodate was added to obtain a final concentration of 0. 02M. The mixture was immediately dispensed into microtubes containing the peptides, mixed and reacted for 30 minutes. 0.02 M ethylene glycol was added to inactivate the remaining sodium periodate. The intermediate Schiff base, formed between the amino terminus of the peptide and the aldehyde formed by oxidation of the carbohydrate portions of HRP was reduced by the addition of 0.2 M sodium borohydride in water. Chromatography of each peptide-peroxidase conjugate was used on Sephadex G25, to remove reagents and excess peptide. In this preferred assay the antibody was used to bind epitopes identified in HIV proteins prepared from lysates of HIV viruses and synthetic HIV peptides, covalently linked to HRP, since the antibody reactive with the HIV epitopes retained a site of antigen binding that could react with the synthetic peptide epitopes bound to the peroxidase.

The granules were coated with HIV proteins as described above. Granules coated with goat anti-HIV IgG were reacted for 24 hours, the granules were washed and reacted with the substrate to determine peroxidase activity and, therefore, peptide binding. With this procedure, only the exact epitopes contained within the synthetic peptide were recognized. In this analysis, the complete repertoire of goat reactivities was identified. Human goat anti-HIV and anti-HIV reactivities were compared, and peptides that reacted only with goat anti-HIV were selected as candidate target epitope targets. These data were not considered including all differences between human and goat reactivity to HIV proteins, since differences between these reactivities and deviations between amino acid peptide sequence homologies in the HIV protein could result in reactivity of missing epitope, secondary for the selection of the synthetic peptide.

EXAMPLE 9 EFFICACY OF ANTIBODY AS A THERAPEUTIC AGENT A study was carried out with a group of immunochemically designed antibodies to determine the efficacy of the antibodies as a therapeutic agent for the treatment of HIV / AIDS. Specifically, a mixture of lysates of HIV isolates HIVIMU, HIV1BAL and HIV2NZ was treated, as in example 3, to eliminate low molecular weight contaminants and HLA class I and class II antigens, and to deglycosylate HIV proteins . The protein mixture was analyzed and found to consist of peptides that replicate or mimic the following regions of HIV1SF2: gpl20 proteins: an epitope region extending from the amino acid residue 4-27 and a second epitope region extending from the residue 54-76 amino acid; gp41: an epitope region extending from amino acid residue 502-531; the p66 / 55 heterodimer of reverse transcriptase: an epitope region extending from residue 254-295 of amino acid; PlO protease: an epitope region extending from the 69-94 amino acid region; Gag gene p24 protein: an epitope region extending from the 166-181 amino acid region; Gag gene pl7 protein: an epitope region extending from the 2-23 amino acid region and a second epitope region extending from the amino acid region 89-122; and Gage gene p7 protein: an epitope region extending from region 390-41 and 438-443 amino acid. These amino acid sequences can not trigger an immune response in humans when contacted by infection or naturally through the environment, but they do elicit an immune response in other mammalian species. The purified and treated proteins were enriched and further purified by using preparative SDS-PAGE electrophoresis and, when desired, by immunoaffinity chromatography using commercially available monoclonal antibodies (ICN Costa Mesa, CA, and Advanced Biotechnologies, Columbia, MD, USA) ), for gpl20, gp41, pp66 / 55, p24, pl7 and plO, each coupled individually to Sepharose CL4B, using procedures known in the art. After purification, each peptide of purified HIV protein, of interest, was individually conjugated to the MDP microparticle which is described in example 4.

The HIV-MDP conjugates were formulated into microparticles as a combination containing equimolar concentrations of each HIV protein / peptide, at a final concentration of approximately 1 mg protein / ml physiological saline. 0.5 ml of HIV / MDP combination was emulsified in 1.5 ml of squalene containing 0. 05% of Tween 80, and was injected subcutaneously in 60 goats in 4 to 6 injection sites per goat. Reinforcement immunizations were given at monthly intervals to obtain the antibody responses. Serum samples of each goat were obtained monthly, before monthly reinforcement, and tested by Western staining, HIV neutralization and enzyme immunoassay, described in example 8.6. Once the desired antibody response was obtained, the goats were subjected to plasmapheresis from the jugular vein using a Baxter A 201-A-401 Autophresis machine. The goat red blood cells were returned to physiological saline. The volume of plasma collected from each animal, in each plasmapheresis, was 350 ml + 5.0 ml. The goat immunological plasma was fractionated with octanoic acid, after adjusting the pH to 4.8 with hydrochloric acid. The mixture was centrifuged and passed through an activated carbon filter to remove the agglomerates and reduce the level of octanoic acid to less than 0.05%. The fraction containing immunoglobulin (IgG) was further purified by passing it through a series of columns: 1. Sephadex G25 with 20 mM phosphate buffer; 2. Ion exchange with Whatman DE53 balanced in phosphate buffer; 3. Sephadex G25 equilibrated in 0.9% saline. The filtrate from the final column was filtered on a sterile filter and then concentrated or diluted to give a final concentration of 10 mg of IgG / ml, with the desired biological activity. SDS-PAGE and Western spotting assays demonstrated goat IgG immunoglobulin reactivity at a dilution of 1: 100 with the seven different viral proteins such as gpl20, gp41, p5l / 66, p24, pl7, p7 and plO. Tests for HIV neutralization demonstrated broad neutralization of laboratory and wild-type HIVl strains, such as those described in Example 8.3. Enzyme-linked immunoassay analysis, described in Example 8.3, demonstrated positive reactions to the nine desired epitopes in sample dilutions of 1: 1000 or more. The production batches satisfying these criteria were given the commercial name of HRG214, as described, and consisted of sterile, pyrogen-free goat IgG at a concentration of 10 mg / ml suspended in 0.9% sodium chloride, without excipients. Each production lot is compared to the reference lot, as described here, to determine the equivalence of the activity of HRG214. The potency is expressed as mg-equivalent / ml. Non-pregnant men and women aged over 18 were selected, based on whether they had a serodiagnosis of HIV infection documented by Western staining, with clinically symptomatic AIDS defining criteria, and with a CD4 T lymphocyte count. 300 cells / ul within 30 days before entering the study. Clinical and laboratory parameters were used to evaluate the efficacy. Clinical parameters included changes in opportunistic infections, changes in body weight, changes in gastrointestinal function, including evacuation consistency and frequency, changes in energy level, changes in appetite, physical strength and endurance, and a general change in the quality of life. Laboratory parameters included changes in CD4 and CD8 lymphocyte numbers, selected hematology, blood chemistry and urinalysis and, when available, changes in viral loads by measuring viral RNA by PCR. The results of the study showed that, with the use of immunochemically designed antibodies, patients improved clinically, with a decrease in opportunistic infections, increase in body weight, changes in gastrointestinal function, including less severe diarrhea, increases in energy level, increase in appetite, improvement in strength and physical endurance, and a general improvement in quality of life. The laboratory parameters showed improvements with increase in the number of CD4 and CD8 lymphocytes, improvement in hematology, blood chemistry and numbers of urine analysis, as well as decrease in viral loads by measuring viral RNA by PCR, and decrease in infectivity, when measured by TCID. The detailed laboratory results of the study are indicated in the annexed tables 91. A 9.13.

EXAMPLE 9.1 CLINICAL EVALUATION OF TOXICITY AND THE EFFICACY OF HRG214 IN THE TREATMENT OF PATIENTS INFECTED WITH HIV Thirty-five patients infected with HIV were treated with HRG214. HRG214 was evaluated for its efficacy in reducing HIV viral load and improving the progression and symptoms of the disease. The patients were stratified by the CD4 / mm3 number (<200 and> 200) the therapeutic regimen: Group 1: HRG214 CD4 < 200, n = 11. Group 2: HRG214 and repeated treatment monthly; CD4 < 200, n = 6. Group 3: HRG214 and repeated treatment to the advance, CD4 < 200, n = 5. Group 4: HRG214 CD4 > 200, n = 7 Group 5: HRG214 and repeated treatment to advance (patients are receiving chemotherapy for malignancy), CD4 < 200, n = 6. Group 6: placebo control CD4 > 500, n = 3. Group 7: Control with placebo, CD4 > 200-500, n = 3. Patients in the treatment groups received 21-28 daily infusions of HRG214 at 1-1.5 mg / kg of body weight. The patients with CD4 were again treated < 200 for three days, monthly (n = 6) or were re-treated with evidence of HIV progression (n = 11). Adverse reactions to minor fever << 2 ° C, chills, headache and muscle pain. Clinical chemistry and hematology measurements during and after treatment remained unchanged or improved in all patients over a period of 90 days. Twenty-eight patients were evaluated for changes in nutritional status. Twenty-four increased from around 1 to 10 kg of body weight, with an average increase of around 2 kg (p = 0.0014). Four of the six patients receiving systemic chemotherapy for malignancy remained stable (n = 2) or lost weight (around 1 and 2.7 kg, respectively). The weight increases were directly correlated with increases in measurements of total serum protein and serum albumin. Quantitative HIV RNA decreased in all treatment groups.

Group 4 HRG214; day 405 315/487 970/1164 • 94% n = 7, s = 7 The rate of loss of CD4 / mm3 over time in all treatment groups was reduced (p <0.01) compared to control groups 6 and 7. A sustained increase in CD4 was observed in groups 2, 3 and 4. Infectivity measurements by microculture (TCID) showed a logarithmic reduction 2 in infectivity by treatment on day 7-14 (p <0.001), which was not obvious from the quantitative HIV-RNA measurements. Clinical changes included increases in appetite and stamina with marked improvements in chronic fatigue syndrome, diarrhea, malabsorption, candidiasis, CMV (excluded retinitis) Herpes simplex and zoster, cutaneous Molluscum contagiosum, oral hairy leukoplakia, wasting syndrome, bacterial folliculitis and pneumonitis, as well as peripheral neuropathy related to HIV. HRG214 offers a new drug that helps the management of HIV infection.

Data for patient groups 2-5 are presented in more detail as follows: Patient group 2: Patients n = 6 Primary objective: To assess the safety and efficacy of HRG214 treatment of HIV infection, at a daily dose of 1.5 mg / kg / day for 28 days and a repeated treatment monthly 3 times. The clinical follow-up will continue for 3 years. Patients will have repeated treatment options with recurrence. Endpoints: The standardization of clinical and laboratory parameters, including improvement in opportunistic infections, incidence of infections, wasting syndrome, peripheral neuropathy and improvement in abnormal blood chemistry and hematology, CD4 and CD8 and reductions in HIV-RNA, they were quantified by means of PCR. The safety variables include: Blood chemistry and hematology and clinical parameters. The efficacy variable includes: quantitative HIV-RNA through CPR, CD4 and CD8 counts. Follow-up period = 3 years Follow-up period to date = > 345 days Demographic study: Patients n = 6. Start date: October 23, 1995; Days passed: 390; survivors = 6; deaths = 0; loss of follow-up = 0. Patient demography: HIV positive; AIDS defining criteria with CD4 number < 50 / mm3. Treatment drug (s): HRG-214 - 1.5 mg / kg / day, with repeated treatments monthly.

TABLE 9.2 GROUP OF PATIENTS 2 TABLE 9.3 GROUP OF PATIENTS 2 TABLE 9.4 GROUP OF PATIENTS 2 Patient group 3: Patients n = 5 Primary objective: To assess the safety and efficacy of treatment with HRG 214 of HIV infection at a daily dose of 1.5 mg / kg / day for 28 days. The clinical follow-up will continue for 3 years. Patients will have options to repeat treatment with recurrence. Endpoints: The standardization of clinical and laboratory parameters including improvements in opportunistic infections, incidence of infections, wasting syndrome, peripheral neuropathy and improvements in blood chemistry and abnormal hematology, CD4 and CD8 and reductions in HIV-RNA, were quantified through CPR. The safety variables include: Blood chemistry and hematology and clinical parameters. The efficacy variable includes: quantitative HIV-RNA through CPR, CD4 and CD8 counts. Follow-up period = 3 years. Follow-up period to date = > 469 days Start date: June 13, 1995; days passed, 380; survivors = 3; deaths = 2, -transfer loss = 0. Patient population: HIV positive patients with defining criteria for AIDS. Five (5) of the five (5) patients had CD4 < 50 / mm3 of blood. HIV-RNA quantified by PCR showed statistically significant reductions after treatment; on day 7 (P = 0.0179) and days 21-28 (P = 0.043). HIV-AFJST measurements on days 60-90 and 120-150 showed reduced but increasing HIV-RNA values. The 5 patients were treated again (three consecutive doses monthly, beginning between days 120-150). After the new treatment, a statistically significant fall (P = 0.0006) was observed in the HIV-RNA measurements on the day > 250. Statistical analyzes were carried out using paired t-test, signed Wilcoxon rank test and Mann-Whitney rank sum test. START DATE: June 13, 1995 FINISHING DATE: open TABLE 9.5 GROUP OF PATIENTS 3 TABLE 9.6 GROUP OF PATIENTS 3 TABLE 9.7 PATIENT GROUP 3 Patient Group 4 Patients n Primary objective: To evaluate the toxicity and efficacy of treatment with HRG214 of HIV infection at a daily dose of 1.5 mg / kg / day for 28 days with IFN inducer on days 1-7 and 21-23 . The clinical follow-up will continue for 3 years. Patients will have the option to repeat the treatment with recurrence. End points: Normalization of clinical parameters and laboratory parameters including improvement in 10, incidence of infections, wasting syndrome, etc., in blood chemistry and abnormal hematology, CD4 and CD8, reductions in HIV-RNA quantified by PCR. Follow-up period: 3 years. Tracking period to date: > 405 days Demography of the study: Patients n = 7; Start date: August 25, 1995; Until the day 390, survivors: 7; deaths: 0, loss of follow-up: 0. Demographics of patients: HIV-positive patients with CD4 number > 200 without defining criterion of AIDS. Treatment drug (s): 1.5 mg / kg / day for 28 days. Patients will have the option to repeat the treatment with recurrence.

TABLE 9.8 PATIENT GROUP 4 TABLE 9.9 GROUP OF PATIENTS 4 TABLE 9.10 GROUP OF PATIENTS 4 Patient group 5. Patients n = 6 Primary objective: To evaluate the toxicity and efficacy of HRG214 treatment. The clinical follow-up will continue for three years. Patients will have options to repeat the treatment with recurrence. End points: Normalization of clinical parameters and laboratory parameters including improvement in 10, incidence of infections, wasting syndrome, etc., in blood chemistry and abnormal hematology, CD4 and CD8, reductions in HIV-RNA quantified by PCR.

Follow-up period: 3 years. Tracking period to date: > 469 days Start date: June 13, 1995; until day 270, survivors = 2; deaths 4 (2 deaths between 180-210, 1 death between 240-270, one death after 270); loss of follow-up: 0. Patient population: HIV positive patients with AIDS defining criteria that include CDR < 200 / mm3 of blood and disseminated Kaposi's sarcoma. Patients were treated with systemic chemotherapy. Two patients died between days 180-210 and two patients died after day 240. HIV-RNA quantified by CPR, showed reductions on days 60-90, 120-150, 180-210 and 240-270 (p = 0.018) . Statistical analysis was performed using paired t-analysis, Wilcox analysis, signed-rank analysis and Mann-Whitney rank-sum analysis.

TABLE 9.11 GROUP OF PATIENTS 5 TABLE 9.12 GROUP OF PATIENTS 5 TABLE 9.13 GROUP OF PATIENTS 5 CD8 / mm3 QUANTII ATIVO TEST Day 1 Day 21-28 Day 60-90 Day 120- Day 180- Day 240- 150 210 270 Media 476.3 303.5 415.8 371 326.4 676 Error of 152.9 62.7 159.1 100.7 96.2 No rule to define Maximum 1170 540 1152 710 618 676 Minimum 98 86 92 62 101 676 Medium 352 271.5 293.5 245 676 358 BIBLIOGRAPHY 1. H. Mitsuya, S. Broder, Nature 325, 773-778 (1987). . The Molecular Biology of Tumor Viruses, J. Tooze et al., Ed. (1973). 3. RNA Tumor Viruses, R. Weiss, Ed. (1982). 4. F. Gonzalez-Scarano, R. E. Shoppe, C. E. Calisher, N. Nathanson, Virology, 120, 42-53 (1982). 5. S. Matsunc, S. Inouye, Infection and Immuni ty, 390, 155-158 (1983). 6. J. Mathews, J. Roehrig, The Journal of Immunology, 129 2763-2767 (1982). 7. M. Robert-Guroff, M. Brown, R. Gallo, Nature 316, 72-74 (1985). 8. R. Weiss, and co-authors, Nature 316, 69-72 (1985). 9. T. Matthews and co-authors, Proceedings of the National Academy of Sciences 83, 9709-9713 (1986). 10. W. Robey and co-authors, Proceedings of the National Academy of Sciences, 83, 7023-7027 (1986). 11. L. Lasky and co-authors, Science 233, 209-212 (1986). 12. D. Zagury and co-authors, Nature 326, 249-250 (1987). 13. J. McDougai and co-authors, Science, 231, 382-385 (1986) 14. S. Putney and co-authors, Science 234, 1392-13905 (1986) 15. S. Norley, R. Kurth, The Retroviridae, J. Levy Ed. (Plenum Press, 1994), volume 5. 16. J. Carison, JAMA 260, 674-679 [1988]. 17. J. Lange and co-authors, Bri tish Medical Journal 292, 228-230 (1986). 18. J. McDougal and co-authors, Journal of Clinical Investigat, 80, 316-324 (1987). 19. Amador A., A. De Rossi, G. Faulker-Valle, 1. Chieco-Bianchi, Clinical Immunology and Immunopathology, 46, 342-351, (1988). 20. A. Amardori and co-authors, The Journal of Imunology, 89, 2146, 2152 (1989). 21. E. Barker, S. W. Barnett, L. Stamataos, J. A. Levy, in The Viruses: The Retroviridae, J. A. Levy Ed. (Plenum Press, New York and London, 1995), volume 4, pages 1-7. 22. J. A. Levy in HIV and the Pathogenesis of AIDS, A. Press, Ed. (ASM Press, Washington, DC, E. U. A., 1994), pages 1-5. 23. D. Nixon, K. Broliden, G. Ogg, P.-A. Broliden, Immunology 76, 515-534 (1992). 24. P. Linsley, J. Ledbetter, E. Kinney-Thomas, S.-L Hu, J Virology 62, 3695-3702 (1988).

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Claims (1)

1. NOVELTY OF THE INVENTION CLAIMS 1. A composition characterized in that it comprises HIV proteins isolated from a lysate of an HIV isolate that has been treated to eliminate the HLA human antigens of class I and class II present in the lysate; wherein the proteins have been deslicosylated and where the proteins comprise at least one epitope region that does not elicit an immune response in man when found by infection or environmental exposure, but unleashes an immune response in at least one non-human mammalian species. 2. - A composition according to claim 1, further characterized in that said epitope region comprises a neutralizing or inactivating region of the HIV protein. 3. - A composition according to claim 1, further characterized in that the epitope region has an amino acid sequence that corresponds to, or immunologically mimics a portion of an amino acid sequence of human protein. 4. - A composition according to claim 1, further characterized in that it has been enriched for said region (s) epitope (s). 5. A composition according to claim 4, further characterized in that the epitope region (s) constitute (n) at least about 25% of the protein. 6. A composition according to claim 5, further characterized in that said epitope region (s) comprises between about 50% and about 95% of the protein. 1 . - A composition according to claim 1, further characterized in that it comprises a mixture of lysates of different HIV isolates. 8. - A composition according to claim 1, further characterized in that it comprises a mixture of lysates of HIVIM, HIV1BA, and HIV2NZ. 9. A composition according to claim 1, further characterized in that the epitope region corresponds to, or mimics at least one protein epitope region of HIV isolate HIV1SF2 that does not elicit an immune response in man when found by infection or environmental exposure, but causes an immune response in at least one other mammalian species. 10. A composition according to claim 9, further characterized by the epitope region corresponding to or mimicking an epitope region of at least one of the following HIV1SF2 proteins: (a) an envelope glycoprotein gpl20 envelope; (b) an enveloping gp41 transmembrane glycoprotein; (c) reverse transcriptase; (d) plO protease; or (e) gag precursor. 11. A composition according to claim 10, further characterized in that at least one of the epitope regions of the HIVSF2 proteins comprises: (a) a region extending from amino acid residue 4 to amino acid residue 27 of the gpl20 glycoprotein; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region that extends from the waste 2 amino acid to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 12. - A composition according to claim 1, further characterized in that it additionally comprises an adjuvant. 13. - A composition according to claim 12, further characterized in that the adjuvant comprises a carrier molecule to which the HIV protein is coupled. 14. A composition according to claim 13, further characterized in that said molecule comprises poly-L-lysine, keyhole limpet hemocyanin, thyroglobulin, an albumin or tetanus toxoid. 15. - A composition according to claim 13, further characterized in that the carrier molecule comprises multiple repeats of a glycopeptide. 16. A composition according to claim 15, further characterized in that the carrier molecule comprises multiple repeats of muramyl dipeptide. 17. A composition according to claim 16, further characterized in that said multiple repeats of the muramyl dipeptide are intertwined. 18. A composition according to claim 17, further characterized in that the multiple repeats of the muramyl dipeptide comprise a terminal dipeptide of L-alanine-D-isoglutamine. 19. A composition, characterized in that it comprises a synthetic peptide comprising an epitope region corresponding to, or mimics a neutralizing or inactivating region of an HIV protein; wherein the peptide does not elicit an immune response in humans when found by infection or environmental exposure, but which does elicit an immune response in at least one non-human mammalian species. 20. - A composition according to claim 19, further characterized in that the epitope region has an amino acid sequence that corresponds to, or mimics a portion of a human protein. 21. A composition according to claim 19, further characterized in that at least one amino acid within the epitope region is modified to increase the MHC interactions or the immunological response obtained after administration of the peptide to a non-human mammal. 22. - A composition according to claim 21, further characterized in that at least one amino acid is modified in order to create an amphipathic helix with the epitope region enclosed between hydrophilic amino acids and hydrophobic amino acids. 23. - A composition according to claim 19, further characterized in that it comprises a mixture of synthetic peptides, wherein the peptides comprise regions of epitope corresponding to, or mimic more than one neutralizing or inactivating region of HIV proteins. 24. - A composition according to claim 19, further characterized in that the epitope region corresponds to or mimics a neutralizing or inactivating region of an HIV isolate protein, HIV1SF2. 25. A composition according to claim 23, further characterized in that the epitope regions correspond to, or mimic more than one neutralizing or inactivating region of the proteins of the HIV isolate, HIV1SF2. 26. A composition according to claim 24 or 25, further characterized in that the HIV1S 2 protein comprises: (a) an envelope glycoprotein outer glycoprotein; (b) an enveloping gp41 transmembrane glycoprotein; (c) reverse transcriptase; (d) plO protease; or (e) gag precursor. 27. A composition according to claim 26, further characterized by the neutralizing or inactivating region of the HIVSF2 protein comprising: (a) a region extending from the amino acid residue 4 to the amino acid residue 27 of the glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein g? 41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region that extends from the waste 2 amino acid to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 28. A composition according to claim 19, further characterized in that it additionally comprises an adjuvant. 29. A composition according to claim 28, further characterized in that the adjuvant comprises a carrier molecule to which the HIV peptide is coupled. 30. A composition according to claim 29., further characterized in that the carrier molecule comprises poly-L-lysine, keyhole limpet hemocyanin, thyroglobulin, an albumin or tetanus toxoid. 31. A composition according to claim 29, further characterized in that the carrier molecule comprises multiple repeats of a glycopeptide. 32. - A composition according to claim 31, further characterized in that the carrier molecule comprises multiple repeats of the muramyl dipeptide. 33. - A composition according to claim 32, further characterized in that the multiple repeats of the muramxlico dipeptide are intertwined. 34. A composition according to claim 33, further characterized in that the multiple repeats of the muramyl dipeptide comprise a terminal dipeptide of L-alanine-D-isoglutamine. 35.- A method to identify a neutralizing or inactivating region of an HIV protein, in which the neutralizing or inactivating region does not trigger an immune response in man when it is found through infection or environmental exposure, but it does provoke an immune response in a non-human mammal; characterized in that said method comprises: (a) extracting HIV proteins from a lysate of an HIV strain; (b) immunizing a non-human mammal with that extract; (c) obtaining antisera from said immunized mammal; (d) employing the antiserum in a competitive immunoassay with human HIV antisera to identify regions of HIV proteins that are recognized by antibodies in said antisera, but not recognized by antibodies in human antisera; and (e) determining which of said regions is a neutralizing or inactivating region. 36. A method according to claim 35, further characterized in that the neutralizing or inactivating region comprises or is homologous to, one of the following regions of an HIV HIV1SF2 isolate protein: (a) (a) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 37.- A method to obtain antibodies that react with an epitope in a neutralizing or inactivating region of an HIV protein, where the neutralizing or inactivating region of the protein can not trigger an immune response in man, when it is found by infection or by environmental exposure, but causes an immune response in a non-human mammal; characterized in said method because it comprises: (a) isolating proteins from a lysate of an HIV isolate; (b) identifying an epitope on at least one of said proteins, having an amino acid sequence corresponding to or mimicking the amino acid sequence of a neutralizing or inactivating region, which can not trigger an immune response in man when it is found by infection or environmental exposure, but elicit an immune response in a non-human mammal; (c) combining said proteins with a physiologically acceptable carrier; (d) immunizing a host non-human mammal, with said proteins and said carrier; Y (e) obtain antibodies for the epitope of the immunized host. A method according to claim 37, further characterized in that the lysate is treated to eliminate HLA class I and class I antigens. 39. A method according to claim 37, further characterized in that the proteins are deglycosylated before being combined with the physiological carrier. 40. A method according to claim 37, further characterized in that the amino acid sequence of the epitope corresponds to, or mimics a portion of a human protein amino acid sequence. 41. - A method according to claim 37, further characterized in that the protein is conjugated with an adjuvant before combining it with a physiologically acceptable carrier. 42. - A method according to claim 41, further characterized in that the adjuvant comprises a macromolecular carrier. 43. - A method according to claim 42, further characterized in that the macromolecular carrier comprises multiple repeats of muramyl dipeptide. 44. A method according to claim 43, further characterized in that the multiple repeats of the muramyl dipeptide comprise a terminal dipeptide of L-alanine-D-isoglutamine. 45. A method according to claim 37, further characterized in that the proteins comprise epitopes that correspond to, or mimic more than one neutralizing or inactivating region. 46. A method according to claim 37, further characterized in that the neutralizing or inactivating region comprises a portion of an envelope glycoprotein or a transmembrane protein. 47. - A method according to claim 45, further characterized in that at least one of the neutralizing or inactivating regions comprises a portion of a envelope glycoprotein or transmembrane protein. 48. A method according to claim 47, further characterized in that it additionally comprises a neutralizing or inactivating region of the plO protease. 49. A method according to claim 37, further characterized in that the epitope corresponds to, or mimics an epitope region of HIV1SF comprising: (a) a region extending from amino acid residue 4 to amino acid residue 27 of the glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region that extends from the waste 2 amino acid to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 50. A method according to claim 49, further characterized in that the proteins comprise epitopes corresponding to or mimic more than one neutralizing or inactivating region and the epitopes correspond to, or mimic two or more of said epitope regions of HIVSF2-51. - A method according to claim 37 or 45, further characterized in that said proteins have been enriched in said epitope (s). 52. A method according to claim 45, further characterized in that the epitopes are present in relative proportions ranging from about 1: 1 to a maximum difference in amount between any two epitopes, of 10: 1. 53. - A method for obtaining antibodies that react with an epitope in a neutralizing or inactivating region of an HIV protein, where the neutralizing or inactivating region of the protein can not trigger an immune response in man when it is found by infection or by environmental exposure , but it does elicit an immune response in a non-human mammal, said method characterized in that it comprises: (a) synthesizing a peptide having an amino acid sequence that corresponds to, or mimics an epitope in a neutralizing or inactivating region of an HIV protein , where the region can not unleash an immune response in humans when it is found through infection or environmental exposure, but causes an immune response in a non-human mammal; (b) combining the peptide with a physiologically acceptable carrier; (c) immunizing a mammalian, non-human host with the peptide and the carrier; and (d) obtaining antibodies to the epitope of said immunized host. 54. A method according to claim 53, further characterized in that the peptide has an amino acid sequence that mimics a portion of an amino acid sequence of human protein. 55. - A method according to claim 3, further characterized in that the peptide is conjugated with an adjuvant before combining it with the physiologically acceptable carrier. 56. A method according to claim 55, further characterized in that the adjuvant comprises a macromolecular carrier. 57. A method according to claim 56, further characterized in that the macromolecular carrier comprises multiple repeats of muramyl dipeptide. 58. A method according to claim 57, further characterized in that the multiple repeats of the muramyl dipeptide comprise a terminal dipeptide of L-alanine-D-isoglutamine. 59. A method according to claim 53, further characterized in that it comprises a mixture of peptides, each of which has an amino acid sequence that corresponds to or mimics an epitope in a neutralizing or inactivating region of an HIV protein; where said region can not unleash an immune response in man, when it is found by infection or environmental exposure, but it does unleash an immune response in a non-human mammal. 60. - A method according to claim 53, further characterized in that the peptide has an amino acid sequence that corresponds to or mimics a neutralizing or inactivating region comprising a portion of an envelope glycoprotein or a transmembrane protein. 61.- A method according to claim 59, further characterized in that at least one of the peptides has an amino acid sequence that corresponds to, or mimics a neutralizing or inactivating region comprising a portion of an envelope glycoprotein or an envelope protein. transmembrane 62. - A method according to claim 61, further characterized in that it additionally comprises a peptide having an amino acid sequence that corresponds to, or mimics a pilo protease neutralizing or inactivating region. 63.- A method according to claim 53, further characterized in that the amino acid sequence of the peptide corresponds to, or mimics the amino acid sequence of an epitope of a neutralizing or activating region of an HIV1SF2 protein, comprising: (a) ) a region extending from the amino acid residue 4 to the amino acid residue 27 of the glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the trans embrana glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 64.- A method according to claim 59, further characterized in that the peptides correspond to, or mimic amino acid sequences of at least two epitopes in neutralizing or inactivating regions of HIV1SF2 proteins; said regions comprising: (a) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to the amino acid residue 76 of the glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 65. - A method according to claim 59, further characterized in that the peptides are present in relative proportions ranging between about 1: 1 and a maximum difference in the amount between any two peptides, of 10: 1. 66.- An antibody that recognizes, and reacts with, an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of an HIV protein, characterized in that said neutralizing or inactivating region of the protein can not elicit an immune response in the man, when it is found by infection or by environmental exposure. 67.- An antibody according to claim 66, further characterized in that the epitope has an amino acid sequence that corresponds to, or immunologically mimics a portion of an amino acid sequence of human protein, in an amino acid sequence of neurotoxin protein. 68.- An antibody according to claim 66, further characterized in that the protein comprises glycoprotein of envelope precursor gplβO, gpl20 devoid of carbohydrate, or transmembrane glycoprotein gp 41. 69. An antibody according to claim 66, further characterized in that the protein comprises a p55 precursor of gag, devoid of carbohydrate, or the p7, p24 or p7 products of gag divided. 70. An antibody according to claim 66, further characterized in that the protein comprises the plO protease devoid of carbohydrate, or the p66 / 55 heterodimer of reverse transcriptase. 71. An antibody according to claim 67, further characterized in that the human protein comprises alpha-photoprotein, asparatyl-protease, nucleotidohydrolase of deoxyuridine 5 ** -triphosphate, cationic eosinophil protein or eosinophil-derived neurotoxin. 72. An antibody according to claim 67, further characterized in that the antibody recognizes an epitope corresponding to or mimics an epitope in one of the following neutralizing or inactivating regions of the HIV isolate, HIV1SF2: (a) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and from amino acid residue 438 to 443 of the p7 protein of the gene gag; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 73. A combination of at least two antibodies, each of which recognizes and reacts with an epitope corresponding to, or mimics an epitope in a neutralizing or inactivating region of an HIV protein; characterized in that the neutralizing or inactivating region of the protenine can not unleash an immune response in man, when it is found by infection or by environmental exposure. 74. A combination of antibodies according to claim 73, further characterized in that each of the antibodies recognizes and reacts with an epitope having an amino acid sequence that corresponds to, or immunologically mimics a portion of a protein amino acid sequence. human, in an amino acid sequence of neurotoxin protein. A combination of antibodies according to claim 73, further characterized in that at least one of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of precursor gpl60 of envelope precursor devoid of carbohydrate, glycoprotein gpl20 transmembrane glycoprotein gp 41. 76.- A combination of antibodies according to claim 73, further characterized in that at least one of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing region or inactivator of a p55 precursor of gag devoid of carbohydrate, or pl7, p24 or p7 products of divided g. 77. - A combination of antibodies according to claim 73, further characterized in that at least one of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of the carbohydrate deprived plO protease, or the heterodimer of reverse transcriptase p66 / 55. 78. A combination of antibodies according to claim 74, further characterized in that the human protein comprises alpha-fetoprotein, aspartyl-protease, 5'-nucleotidehydrolase-deoxyuridinetriphosphate, cationic eosinophilic protein or eosinophil-derived neurotoxin. 79. A combination of antibodies according to claim 73, further characterized in that at least one of the antibodies recognizes an epitope corresponding to or mimics an epitope in one of the following neutralizing or inactivating regions of the HIV isolate HIVISF ?: (a) ) a region extending from the amino acid residue 4 to the amino acid residue 27 of the glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene protein • gag p24; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region that extends from the waste 2 amino acid to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 80. A combination of antibodies according to claim 79, further characterized in that it comprises antibodies that recognize an epitope corresponding to or mimic an epitope in each of the following neutralizing or inactivating regions of the HIV isolate HIVISF ?: (a) a region extending from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; and (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41. 81. A combination of antibodies according to claim 79, further characterized in that it comprises antibodies that recognize an epitope corresponding to or mimic an epitope in each of the following neutralizing or inactivating regions of HIV isolate HIVlsF2: (a) a region extending from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41; and (d) a region extending from amino acid residue 69 to 94 of the plO protease. 82. A combination of antibodies according to claim 79, further characterized in that it comprises antibodies that recognize epitopes corresponding to or mimicking epitopes in each of the following neutralizing or inactivating regions of the HIV isolate HIV1SF2: (a) a region that extends from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41; and at least one of the following neutralizing or inactivating regions: (d) a region extending from the amino acid residue 69 through 94 of the plO protease; (e) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (f) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gene gag; (g) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (h) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein; (i) a region extending from amino acid residue 254 to 295 of the reverse transcriptase heterodimer p66 / 55. 83. A combination of antibodies according to claim 73, further characterized in that the antibodies recognize epitopes corresponding to or mimic epitopes in the following neutralizing or inactivating regions of the HIV isolate. HIV1SF2. "(A) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to the amino acid residue 76 of the glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 84. - A composition characterized in that it comprises a combination of antibodies according to claim 79, in a pharmaceutically acceptable carrier. 85.- A composition characterized in that it comprises a combination of antibodies according to claim 80, in a pharmaceutically acceptable carrier. 86.- A composition characterized in that it comprises a combination of antibodies according to claim 81, in a pharmaceutically acceptable carrier. 87. A composition characterized in that it comprises a combination of antibodies according to claim 82, in a pharmaceutically acceptable carrier. 88. A composition characterized in that it comprises a combination of antibodies according to claim 83, in a pharmaceutically acceptable carrier. 89. An antibody according to claim 66, further characterized in that it is bound to a toxin or a radioactive material. 90. An antibody according to claim 66, further characterized in that it is an aggregate with a human T cell activator. 91.- A composition, characterized in that it comprises an antibody according to claim 66, in combination with asida-3? -deoxythidine, 2 ', 3'-dideoxycytidine, 2', 3'-dideoxy-2 ', 3' -dideshydrocytidine. 92.- A composition, characterized in that it comprises the proteins of claim 1, in combination with a pharmaceutically acceptable carrier. 93.- A composition according to claim I, characterized in that the proteins are coupled to a macromolecular carrier. 94. A composition according to claim 93, further characterized by the carrier being a microparticle of muramyl dipeptide. 95.- A composition characterized in that it comprises one or more synthetic peptides of claim 19, in combination with a pharmaceutically acceptable carrier. 96. A composition according to claim 95, further characterized in that the peptides are coupled to a macromolecular carrier. 97. - A composition according to claim 96, further characterized in that the carrier is a microparticle of muramyl dipeptide. 98.- The use of a composition comprising one or more antibodies, each of which recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of an HIV protein, where the neutralizing or inactivating region of the protein can not trigger an immune response in man, for the manufacture of a drug to inhibit an HIV infection in a human infected with the virus. 99. The use according to claim 98, further characterized in that the epitope has an amino acid sequence that corresponds to or immunologically imitates a portion of an amino acid sequence of human protein, when found by infection or environmental exposure. 100. The use according to claim 98, further characterized in that the protein comprises the envelope precursor devoid of carbohydrate gpl60, glycoprotein gpl20 or transmembrane gp 41. 101. The use according to claim 99, further characterized in that the protein comprises the precursor of gag p55 devoid of carbohydrate, or the products of gag divided pl7, p24 or p7. 102. The use according to claim 98, further characterized in that the protein comprises plO protease devoid of carbohydrate or the reverse transcriptase heterodimer p66 / 55. 103. The use according to claim 98, further characterized in that the medicament obtained with said antibodies provides about 0.1 to 200 mg of said antibodies to the patient, per day. 104. The use according to claim 98, further characterized in that when a combination of antibodies is administered, the ratio of each antibody with respect to the others, does not differ by more than a factor of 10. 105.- The use of according to claim 104, further characterized in that the ratio of each of the antibodies, one with respect to the other, is approximately 1: 1. 106. - The use according to claim 98, further characterized in that the antibodies are conjugated to a macromolecular carrier. 107. The use according to claim 106, further characterized in that the carrier is a microparticle of muramyl dipeptide. 108. The use according to claim 98, further characterized in that the antibody recognizes an epitope corresponding to, or mimics an epitope in one of the following neutralizing or isolating regions of the HIV isolate HIV isolate: (a) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to the amino acid residue 76 of the glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region that extends from the waste 2 amino acid to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 109. The use according to claim 108, further characterized in that at least two antibodies are administered and each of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope of a neutralizing or inactivating region of an HIV protein.; wherein the neutralizing or inactivating region of the protein can not unleash an immune response in man when it is found by infection or by environmental exposure. 110. The use according to claim 109, further characterized in that at least one of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of the envelope precursor devoid of carbohydrate gpl60, the glycoprotein gpl20 or the transmembrane gp41. 111. The use according to claim 109, further characterized in that at least one of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of the p55 precursor of gag. * devoid of carbohydrate, or the products of gag divided pl7, p24 or p7. 112. The use according to claim 109, further characterized in that at least one of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of the carbohydrate deprived plO protease, or the heterodimer of reverse transcriptase p66 / 55. 113. The use according to claim 109, further characterized in that it comprises antibodies that recognize and react with epitopes corresponding to or mimic the epitopes of each of the following neutralizing or inactivating regions of the HIV isolate HIV1SF2: (a) a region which extends from the amino acid residue 4 to the amino acid residue 27 of the glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 114. The use of a composition comprising one or more antibodies, each of which recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of an HIV protein, wherein the neutralizing region or protein inactivator can not unleash an immune response in man, for the manufacture of a drug to neutralize or inactivate one or more essential steps in the life cycle of HIV, in a patient infected with the virus. The use according to claim 112, further characterized in that the epitope has an amino acid sequence that corresponds to or that immunologically mimics a portion of an amino acid sequence of human protein. The use according to claim 114, further characterized in that the antibody recognizes an epitope corresponding to or mimics an epitope in one of the following neutralizing or inactivating regions of the HIV isolate HIV1SF2: (a) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 117. The use according to claim 115, further characterized in that at least two antibodies are administered and each of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of an HIV protein. , where the neutralizing or inactivating region of the protein can not trigger an immune response in man. 118. The use according to claim 114, further characterized in that it comprises antibodies that recognize and react with epitopes from each of the following neutralizing or inactivating regions of the HIV isolate HIVISF: (a) a region extending from the residue of amino acid 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; and (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41. The use according to claim 114, further characterized in that it comprises antibodies that recognize and react with epitopes corresponding to or mimic the epitopes of each of the following neutralizing or inactivating regions of the HIV isolate HIV1SF2: (a) a region extending from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41; and (d) a region extending from amino acid residue 69 to 94 of the plO protease. The use according to claim 114, further characterized in that it comprises antibodies that recognize and react with epitopes corresponding to or mimic epitopes of each of the following neutralizing or inactivating regions of the HIV isolate HIV1SF2: (a) a region that extends from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region that extends * from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 121. The use according to claim 114, further characterized in that the antibodies are conjugated to a macromolecular carrier. 122. The use according to claim 121, further characterized in that the carrier comprises a muramyl dipeptide microparticle. 123.- The use of a composition comprising one or more antibodies, each of which recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of an HIV protein, wherein the neutralizing region or protein inactivator can not unleash an immune response in man when found by infection or by environmental exposure; for the manufacture of a medicament for preventing HIV infection in a patient who has been exposed to HIV. 124. The use according to claim 119, further characterized in that the epitope has an amino acid sequence that corresponds to or immunologically mimics a portion of a human protein amino acid sequence. 125. The use according to claim 119, further characterized in that the antibody recognizes an epitope corresponding to or mimics an epitope in one of the following neutralizing or inactivating regions of the HIV isolate HIV1SF2: (a) a region extending from amino acid residue 4 to amino acid residue 27 of glycoprotein gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of glycoprotein gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of the transmembrane glycoprotein gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p66 / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 126. The use according to claim 125, further characterized in that at least two antibodies are administered, and each of the antibodies recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of a protein. HIV, where the neutralizing or inactivating region of said protein can not trigger an immune response in man. 127. The use according to claim 125, further characterized in that it comprises antibodies that recognize and react with epitopes corresponding to or mimic epitopes of each of the following neutralizing or inactivating regions of the HIV isolate HIVINF ?: (a) a region extending from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; and (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41. 128. The use according to claim 125, further characterized in that it comprises antibodies that recognize and react with epitopes corresponding to or mimic epitopes of each of the following neutralizing or inactivating regions of the HIV isolate HIVINF ?: (a) a region extending from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41; and (d) a region extending from amino acid residue 69 to 94 of the plO protease. 129. The use according to claim 125, further characterized in that it comprises antibodies that recognize and react with epitopes corresponding to or mimic epitopes of each of the following neutralizing or inactivating regions of the HIV isolate HIVISF ?: (a) a region extending from amino acid residue 4 to amino acid residue 27 of gpl20; (b) a region extending from amino acid residue 54 to amino acid residue 76 of gpl20; (c) a region extending from amino acid residue 502 to amino acid residue 541 of gp41; (d) a region extending from amino acid residue 254 to amino acid residue 295 of the reverse transcriptase heterodimer p6β / 55; (e) a region extending from amino acid residue 69 to 94 of the plO protease; (f) a region extending from amino acid residue 166 to amino acid residue 181 of the gene gag p24 protein; (g) a region extending from amino acid residue 390 to amino acid residue 410, and amino acid residue 438 to 443 of the p7 protein of the gag gene; (h) a region extending from amino acid residue 2 to amino acid residue 23 of the gene gag pl7 protein; or (i) a region extending from amino acid residue 89 to amino acid residue 122 of the gene gag pl7 protein. 130.- A method to detect the presence of HIV in a sample of biological fluid, from an individual who may have been infected with HIV, characterized in that it comprises using an antibody of claim 66 in an antibody-antigen analysis in which the antibody is combined with a sample of the body of the individual, under conditions that lead to the formation of antibody-antigen complex, and determine whether the antibody binds to an HIV antigen. 131.- A method to detect the presence of HIV in an individual who may have been infected with HIV, characterized in that it comprises employing an antibody of claim 66 in an enzyme immunoassay, wherein the antibody is conjugated to an enzyme and contacted with an individual's body fluid sample, under conditions that lead to the formation of the antibody complex -antigen, and determine if the antibody binds to an HIV antigen. 132. A method for purifying protein that contains at least one epitope corresponding to or that mimics an epitope of a neutralizing or inactivating region of an HIV protein, from a protein solution, characterized in that it comprises immobilizing an antibody according to the Claim 66, to a solid substrate or support; contacting the immobilized antibody with a solution containing the protein under conditions suitable for the formation of immunological complexes between the antibody and the protein; Separate the unbound protein from the protein bound to the antibody and release the antibody protein protein and recover said protein. 133. A composition characterized in that it comprises viral proteins isolated from a viral lysate that has been treated to eliminate the antigens of human HLA class I and class II, present in the lysate, where the proteins have been deglycosylated and where the proteins comprise at least less an epitope region that does not elicit an immune response in man when it is found through infection or environmental exposure, but which elicits an immune response in at least one non-human mammalian species. 134. A composition according to claim 133, further characterized in that the epitope region comprises a neutralizing or inactivating region of the viral protein. 135. A composition according to claim 133, further characterized in that the epitope region has an amino acid sequence that corresponds to or that immunologically mimics a portion of an amino acid sequence of human protein. 136.- A composition according to claim 133, further characterized in that the proteins are coupled to a macromolecular carrier. 137. - A composition according to claim 136, further characterized in that the carrier is a microparticle of muramyl dipeptide. 138.- A composition according to claim 136, further characterized in that the muramyl dipeptide comprises a terminal dipeptide of L-alanine-D-isoglutamine. 139. A composition characterized in that it comprises a synthetic peptide comprising an epitope region corresponding to or that mimics a neutralizing or inactivating region of a viral protein; where the peptide does not elicit an immune response in humans when found by infection or environmental exposure, but does elicit an immune response in at least one non-human mammal. A composition according to claim 139, further characterized in that the epitope region has an amino acid sequence that corresponds to or that immunologically mimics a portion of a human protein amino acid sequence. 141. A composition according to claim 139, further characterized in that the proteins are coupled to a macromolecular carrier. 142. - A composition according to claim 141, further characterized in that the carrier is a microparticle of muramyl dipeptide. 143. A composition according to claim 141, further characterized in that the muramyl dipeptide comprises a terminal dipeptide of L-alanine-D-isoglutamine. 144.- A method to identify a neutralizing or inactivating region of a viral protein, where the neutralizing or inactivating region does not trigger an immune response in man when it is found by infection or environmental exposure, but it does provoke an immune response in an animal non-human; characterized in that said method comprises: (a) extracting viral proteins from a viral lysate; (b) immunizing a non-human mammal with the extract; (c) obtaining antisera from the immunized mammal; (d) employing the antisera in a competitive immunoassay with human viral antisera, to identify regions of viral proteins that are recognized by antibodies present in antisera, but not recognized by the antibodies of human sera; and (e) determining which of said regions is a neutralizing or inactivating region. 145. - A method for obtaining antibodies that react with an epitope in a neutralizing or inactivating region of a protein, where the neutralizing or inactivating region of the protein can not trigger an immune response in man when it is found through infection or environmental exposure, but it does elicit an immunological response in a non-human mammal, characterized in that method because it comprises: (a) isolating proteins from a viral lysate; (b) identifying an epitope on at least one of the proteins having an amino acid sequence corresponding to or mimicking the amino acid sequence of a neutralizing or inactivating region which can not trigger an immune response in man, but which elicits a response immunological in a non-human mammal; (c) combining said proteins with a physiologically acceptable carrier; (d) immunizing a non-human mammalian host with the proteins and the carrier; and (e) obtaining antibodies to the epitope of said immunized host. 146. A method according to claim 145, further characterized in that the proteins are treated to eliminate HLA class I and class II antigens and to eliminate carbohydrates before combining with the physiological carrier. 147. - A method according to claim 146, further characterized in that the proteins are conjugated to a macromolecular carrier comprising a microparticle of muramyl dipeptide, before combining them with the physiological carrier. 148.- A method to obtain antibodies that react with an epitope in a neutralizing or inactivating region of a viral protein, where the neutralizing or inactivating region of the protein can not trigger an immune response in humans when it is found through infection or exposure environmental, but causes an immune response in a non-human mammal; characterized in said method because it comprises: (a) synthesizing a peptide having an amino acid sequence that corresponds to or mimics an epitope in a neutralizing or inactivating region of a viral protein; where the region can not unleash an immune response in man but elicits an immune response in a non-human mammal; (b) combining the peptide with a physiologically acceptable carrier; (c) immunizing a non-human mammalian host with the peptide and the carrier; and (d) obtaining antibodies to the epitope from the immunized host. 149. - A method according to claim 148, further characterized in that the proteins are treated to eliminate HLA class I and class III antigens and to eliminate the carbohydrates before combining them with the physiological carrier. 150. A method according to claim 149, further characterized in that the proteins are conjugated to a macromolecular carrier comprising a muramyl dipeptide microparticle before combining them with the physiological carrier. 151. An antibody that recognizes and reacts with, an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of a viral protein, where the neutralizing or inactivating region of the protein can not trigger an immune response in man, when is found by infection or environmental exposure. 152. A combination of at least two antibodies, characterized in that each of them recognizes and reacts with, an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of a viral protein; where the neutralizing or inactivating region of the protein can not unleash an immune response in man when it is found through infection or environmental exposure. 153. The use of a composition comprising one or more antibodies, each of which recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of a virus protein, where the neutralizing or inactivating region of the protein can not unleash an immune response in man when it is found by infection or by environmental exposure; for the manufacture of a medicament for inhibiting a viral infection in a human infected with the virus. 154. The use according to claim 153, further characterized in that the antibodies are conjugated to muramyl dipeptide microparticles. 155.- The use of a composition comprising one or more antibodies, each of which recognizes and reacts with an epitope corresponding to or mimics an epitope in a neutralizing or inactivating region of an HIV protein, where the neutralizing or inactivating region of the protein can not unleash an immune response in man when it is found by infection or by environmental exposure; for the manufacture of a medicament for preventing a viral infection in a patient who has been exposed to the virus. 156.- A method for detecting the presence of a virus in a sample of biological fluid of an individual, which may have been infected with the virus, characterized in that it comprises: using an antibody of claim 150 in an antibody-antigen analysis in that the antibody is combined with a sample of fluid from the body of the individual, under conditions that lead to the formation of the antibody-antigen complex; and determine whether the antibody binds to a virus antigen. 157.- A method for detecting the presence of a virus in an individual that may have been infected with the virus, characterized in that it comprises using an antibody of claim 151 in an enzyme immunoassay; where the antibody is conjugated to an enzyme and left in contact with a sample of fluid from the body of the individual, under conditions that lead to the formation of the antibody-antigen complex; and determine whether the antibody binds to a virus antigen. 158. A method for purifying protein containing at least one epitope corresponding to or mimicking an epitope of a neutralizing or inactivating region of a viral protein, from a protein solution, characterized in that it comprises immobilizing an antibody according to the claim 150, on a solid substrate or support; contacting the immobilized antibody with a solution containing the protein, under conditions suitable for the formation of immunological complexes between the antibody and the protein; separating the unbound protein from the protein bound to the antibody; and releasing the bound protein from the antibody and recovering said protein.