CN1173735C - Use of anti-gp 39 antibodies for treatment and/or reversal of lupus and associated kidney disease - Google Patents
Use of anti-gp 39 antibodies for treatment and/or reversal of lupus and associated kidney disease Download PDFInfo
- Publication number
- CN1173735C CN1173735C CNB998067563A CN99806756A CN1173735C CN 1173735 C CN1173735 C CN 1173735C CN B998067563 A CNB998067563 A CN B998067563A CN 99806756 A CN99806756 A CN 99806756A CN 1173735 C CN1173735 C CN 1173735C
- Authority
- CN
- China
- Prior art keywords
- cell
- antibody
- cd40cr
- mice
- treatment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- 206010025135 lupus erythematosus Diseases 0.000 title claims abstract description 21
- 208000017169 kidney disease Diseases 0.000 title claims abstract description 9
- 238000011282 treatment Methods 0.000 title abstract description 47
- 239000012634 fragment Substances 0.000 claims abstract description 21
- 230000002441 reversible effect Effects 0.000 claims abstract description 8
- 101150013553 CD40 gene Proteins 0.000 claims description 66
- 102100040245 Tumor necrosis factor receptor superfamily member 5 Human genes 0.000 claims description 65
- 230000004927 fusion Effects 0.000 claims description 29
- 102100032937 CD40 ligand Human genes 0.000 claims description 18
- 206010001580 Albuminuria Diseases 0.000 claims description 14
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 12
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 11
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 claims description 10
- 230000003172 anti-dna Effects 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 7
- 201000008383 nephritis Diseases 0.000 claims description 6
- 230000006378 damage Effects 0.000 claims description 4
- 210000003734 kidney Anatomy 0.000 claims description 3
- 108010029697 CD40 Ligand Proteins 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims 6
- 238000000034 method Methods 0.000 abstract description 33
- 201000010099 disease Diseases 0.000 abstract description 17
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 17
- 210000003719 b-lymphocyte Anatomy 0.000 description 77
- 210000004027 cell Anatomy 0.000 description 63
- 102100035920 Probable hydrolase PNKD Human genes 0.000 description 61
- 241000699670 Mus sp. Species 0.000 description 57
- 108090000623 proteins and genes Proteins 0.000 description 44
- 235000018102 proteins Nutrition 0.000 description 42
- 102000004169 proteins and genes Human genes 0.000 description 42
- 230000004913 activation Effects 0.000 description 41
- 101000868215 Homo sapiens CD40 ligand Proteins 0.000 description 16
- 108090000978 Interleukin-4 Proteins 0.000 description 16
- 102000004388 Interleukin-4 Human genes 0.000 description 16
- 239000000427 antigen Substances 0.000 description 16
- 108091007433 antigens Proteins 0.000 description 16
- 102000036639 antigens Human genes 0.000 description 16
- 229940028885 interleukin-4 Drugs 0.000 description 16
- 201000000596 systemic lupus erythematosus Diseases 0.000 description 16
- 238000002560 therapeutic procedure Methods 0.000 description 15
- 101000759376 Escherichia phage Mu Tail sheath protein Proteins 0.000 description 14
- 230000000694 effects Effects 0.000 description 14
- 238000002474 experimental method Methods 0.000 description 14
- 238000005516 engineering process Methods 0.000 description 13
- 210000002443 helper t lymphocyte Anatomy 0.000 description 13
- 238000002360 preparation method Methods 0.000 description 13
- 241000699800 Cricetinae Species 0.000 description 12
- 108060003951 Immunoglobulin Proteins 0.000 description 12
- 241000699666 Mus <mouse, genus> Species 0.000 description 12
- 102000018358 immunoglobulin Human genes 0.000 description 12
- 238000001114 immunoprecipitation Methods 0.000 description 12
- 238000011160 research Methods 0.000 description 12
- 230000014509 gene expression Effects 0.000 description 11
- 239000003446 ligand Substances 0.000 description 11
- 239000013612 plasmid Substances 0.000 description 11
- 238000002347 injection Methods 0.000 description 10
- 239000007924 injection Substances 0.000 description 10
- 108020003175 receptors Proteins 0.000 description 10
- 102000005962 receptors Human genes 0.000 description 10
- 230000004083 survival effect Effects 0.000 description 10
- 238000012360 testing method Methods 0.000 description 10
- 210000001744 T-lymphocyte Anatomy 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 102000000743 Interleukin-5 Human genes 0.000 description 8
- 108010002616 Interleukin-5 Proteins 0.000 description 8
- 230000022131 cell cycle Effects 0.000 description 8
- 208000023275 Autoimmune disease Diseases 0.000 description 7
- 238000010168 coupling process Methods 0.000 description 7
- 238000005859 coupling reaction Methods 0.000 description 7
- 210000004408 hybridoma Anatomy 0.000 description 7
- 238000002372 labelling Methods 0.000 description 7
- 238000000746 purification Methods 0.000 description 7
- 230000001225 therapeutic effect Effects 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 241000725303 Human immunodeficiency virus Species 0.000 description 6
- 206010020751 Hypersensitivity Diseases 0.000 description 6
- 208000026935 allergic disease Diseases 0.000 description 6
- 230000007815 allergy Effects 0.000 description 6
- 239000002299 complementary DNA Substances 0.000 description 6
- 230000004069 differentiation Effects 0.000 description 6
- 201000004997 drug-induced lupus erythematosus Diseases 0.000 description 6
- 230000002163 immunogen Effects 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 6
- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 6
- 230000008569 process Effects 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 239000007787 solid Substances 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 230000000890 antigenic effect Effects 0.000 description 5
- 238000004043 dyeing Methods 0.000 description 5
- 229940088598 enzyme Drugs 0.000 description 5
- 230000036039 immunity Effects 0.000 description 5
- 239000002773 nucleotide Substances 0.000 description 5
- 125000003729 nucleotide group Chemical group 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 239000003053 toxin Substances 0.000 description 5
- 231100000765 toxin Toxicity 0.000 description 5
- 108700012359 toxins Proteins 0.000 description 5
- IQFYYKKMVGJFEH-XLPZGREQSA-N Thymidine Chemical compound O=C1NC(=O)C(C)=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 IQFYYKKMVGJFEH-XLPZGREQSA-N 0.000 description 4
- 230000003213 activating effect Effects 0.000 description 4
- 230000000340 anti-metabolite Effects 0.000 description 4
- 229940100197 antimetabolite Drugs 0.000 description 4
- 239000002256 antimetabolite Substances 0.000 description 4
- 150000001720 carbohydrates Chemical class 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 231100000673 dose–response relationship Toxicity 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 230000003053 immunization Effects 0.000 description 4
- 238000002649 immunization Methods 0.000 description 4
- ICIWUVCWSCSTAQ-UHFFFAOYSA-M iodate Chemical compound [O-]I(=O)=O ICIWUVCWSCSTAQ-UHFFFAOYSA-M 0.000 description 4
- 150000002632 lipids Chemical class 0.000 description 4
- 230000005855 radiation Effects 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- 238000011287 therapeutic dose Methods 0.000 description 4
- 208000030507 AIDS Diseases 0.000 description 3
- 229920000936 Agarose Polymers 0.000 description 3
- 206010003445 Ascites Diseases 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 3
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 3
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 3
- 102000013275 Somatomedins Human genes 0.000 description 3
- 229940122803 Vinca alkaloid Drugs 0.000 description 3
- 229940100198 alkylating agent Drugs 0.000 description 3
- 239000002168 alkylating agent Substances 0.000 description 3
- 230000001028 anti-proliverative effect Effects 0.000 description 3
- 230000005875 antibody response Effects 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 238000012790 confirmation Methods 0.000 description 3
- 235000018417 cysteine Nutrition 0.000 description 3
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 3
- 230000008021 deposition Effects 0.000 description 3
- 230000029087 digestion Effects 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 239000007943 implant Substances 0.000 description 3
- 238000004255 ion exchange chromatography Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 230000036470 plasma concentration Effects 0.000 description 3
- 229910052697 platinum Inorganic materials 0.000 description 3
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000008521 reorganization Effects 0.000 description 3
- 206010039073 rheumatoid arthritis Diseases 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- 206010002198 Anaphylactic reaction Diseases 0.000 description 2
- 230000003844 B-cell-activation Effects 0.000 description 2
- 108010021064 CTLA-4 Antigen Proteins 0.000 description 2
- 102000008203 CTLA-4 Antigen Human genes 0.000 description 2
- 229940045513 CTLA4 antagonist Drugs 0.000 description 2
- 241001269238 Data Species 0.000 description 2
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 2
- 102100022339 Integrin alpha-L Human genes 0.000 description 2
- 108010064593 Intercellular Adhesion Molecule-1 Proteins 0.000 description 2
- 102100037877 Intercellular adhesion molecule 1 Human genes 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- 108010064548 Lymphocyte Function-Associated Antigen-1 Proteins 0.000 description 2
- 108700018351 Major Histocompatibility Complex Proteins 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 241000283973 Oryctolagus cuniculus Species 0.000 description 2
- 108090000526 Papain Proteins 0.000 description 2
- 108010004729 Phycoerythrin Proteins 0.000 description 2
- 239000002202 Polyethylene glycol Substances 0.000 description 2
- 239000004365 Protease Substances 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 2
- 230000000172 allergic effect Effects 0.000 description 2
- 230000036783 anaphylactic response Effects 0.000 description 2
- 208000003455 anaphylaxis Diseases 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000005784 autoimmunity Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000027455 binding Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 230000007910 cell fusion Effects 0.000 description 2
- 239000003638 chemical reducing agent Substances 0.000 description 2
- 230000006957 competitive inhibition Effects 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 239000003937 drug carrier Substances 0.000 description 2
- 230000007717 exclusion Effects 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- 239000000945 filler Substances 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 208000033065 inborn errors of immunity Diseases 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000003993 interaction Effects 0.000 description 2
- 238000010255 intramuscular injection Methods 0.000 description 2
- 239000007927 intramuscular injection Substances 0.000 description 2
- 238000007913 intrathecal administration Methods 0.000 description 2
- 238000010253 intravenous injection Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000002502 liposome Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 230000000527 lymphocytic effect Effects 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000004005 microsphere Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000000214 mouth Anatomy 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 229940055729 papain Drugs 0.000 description 2
- 235000019834 papain Nutrition 0.000 description 2
- 239000008194 pharmaceutical composition Substances 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 208000028529 primary immunodeficiency disease Diseases 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 238000010188 recombinant method Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000004062 sedimentation Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- 238000010254 subcutaneous injection Methods 0.000 description 2
- 239000007929 subcutaneous injection Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 230000020382 suppression by virus of host antigen processing and presentation of peptide antigen via MHC class I Effects 0.000 description 2
- 230000000699 topical effect Effects 0.000 description 2
- 238000001890 transfection Methods 0.000 description 2
- 238000003146 transient transfection Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- 230000003612 virological effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- OOIBFPKQHULHSQ-UHFFFAOYSA-N (3-hydroxy-1-adamantyl) 2-methylprop-2-enoate Chemical compound C1C(C2)CC3CC2(O)CC1(OC(=O)C(=C)C)C3 OOIBFPKQHULHSQ-UHFFFAOYSA-N 0.000 description 1
- RNAMYOYQYRYFQY-UHFFFAOYSA-N 2-(4,4-difluoropiperidin-1-yl)-6-methoxy-n-(1-propan-2-ylpiperidin-4-yl)-7-(3-pyrrolidin-1-ylpropoxy)quinazolin-4-amine Chemical compound N1=C(N2CCC(F)(F)CC2)N=C2C=C(OCCCN3CCCC3)C(OC)=CC2=C1NC1CCN(C(C)C)CC1 RNAMYOYQYRYFQY-UHFFFAOYSA-N 0.000 description 1
- 208000029483 Acquired immunodeficiency Diseases 0.000 description 1
- 208000035285 Allergic Seasonal Rhinitis Diseases 0.000 description 1
- 108091008875 B cell receptors Proteins 0.000 description 1
- 208000037914 B-cell disorder Diseases 0.000 description 1
- DWRXFEITVBNRMK-UHFFFAOYSA-N Beta-D-1-Arabinofuranosylthymine Natural products O=C1NC(=O)C(C)=CN1C1C(O)C(O)C(CO)O1 DWRXFEITVBNRMK-UHFFFAOYSA-N 0.000 description 1
- 102100027207 CD27 antigen Human genes 0.000 description 1
- 102000017420 CD3 protein, epsilon/gamma/delta subunit Human genes 0.000 description 1
- 108050005493 CD3 protein, epsilon/gamma/delta subunit Proteins 0.000 description 1
- 101150098822 CD53 gene Proteins 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 108010062580 Concanavalin A Proteins 0.000 description 1
- 102100039498 Cytotoxic T-lymphocyte protein 4 Human genes 0.000 description 1
- 238000011767 DBA/2J (JAX™ mouse strain) Methods 0.000 description 1
- 108700003861 Dominant Genes Proteins 0.000 description 1
- 102100025137 Early activation antigen CD69 Human genes 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 206010018364 Glomerulonephritis Diseases 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 101000914511 Homo sapiens CD27 antigen Proteins 0.000 description 1
- 101000889276 Homo sapiens Cytotoxic T-lymphocyte protein 4 Proteins 0.000 description 1
- 101000934374 Homo sapiens Early activation antigen CD69 Proteins 0.000 description 1
- 101001059454 Homo sapiens Serine/threonine-protein kinase MARK2 Proteins 0.000 description 1
- 101000716102 Homo sapiens T-cell surface glycoprotein CD4 Proteins 0.000 description 1
- 101000914514 Homo sapiens T-cell-specific surface glycoprotein CD28 Proteins 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 1
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 1
- 102000000588 Interleukin-2 Human genes 0.000 description 1
- 108010002350 Interleukin-2 Proteins 0.000 description 1
- 208000003456 Juvenile Arthritis Diseases 0.000 description 1
- 206010059176 Juvenile idiopathic arthritis Diseases 0.000 description 1
- 206010050551 Lupus-like syndrome Diseases 0.000 description 1
- 208000034578 Multiple myelomas Diseases 0.000 description 1
- 108091008604 NGF receptors Proteins 0.000 description 1
- 108010032605 Nerve Growth Factor Receptors Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 241000276498 Pollachius virens Species 0.000 description 1
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 108090000412 Protein-Tyrosine Kinases Proteins 0.000 description 1
- 102000004022 Protein-Tyrosine Kinases Human genes 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 206010038997 Retroviral infections Diseases 0.000 description 1
- 239000006146 Roswell Park Memorial Institute medium Substances 0.000 description 1
- 239000012722 SDS sample buffer Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 206010039710 Scleroderma Diseases 0.000 description 1
- 206010048908 Seasonal allergy Diseases 0.000 description 1
- 102100028904 Serine/threonine-protein kinase MARK2 Human genes 0.000 description 1
- 208000021386 Sjogren Syndrome Diseases 0.000 description 1
- 230000006044 T cell activation Effects 0.000 description 1
- 102100036011 T-cell surface glycoprotein CD4 Human genes 0.000 description 1
- 102100027213 T-cell-specific surface glycoprotein CD28 Human genes 0.000 description 1
- 102100033725 Tumor necrosis factor receptor superfamily member 16 Human genes 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000002052 anaphylactic effect Effects 0.000 description 1
- 238000011091 antibody purification Methods 0.000 description 1
- 238000009175 antibody therapy Methods 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- IQFYYKKMVGJFEH-UHFFFAOYSA-N beta-L-thymidine Natural products O=C1NC(=O)C(C)=CN1C1OC(CO)C(O)C1 IQFYYKKMVGJFEH-UHFFFAOYSA-N 0.000 description 1
- 230000003851 biochemical process Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 208000019069 chronic childhood arthritis Diseases 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000024203 complement activation Effects 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 239000003636 conditioned culture medium Substances 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000000586 desensitisation Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 239000012636 effector Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 238000001400 expression cloning Methods 0.000 description 1
- 239000011536 extraction buffer Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000012757 fluorescence staining Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000011207 functional examination Methods 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 210000005260 human cell Anatomy 0.000 description 1
- 230000005934 immune activation Effects 0.000 description 1
- 208000026278 immune system disease Diseases 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 230000009851 immunogenic response Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 230000001861 immunosuppressant effect Effects 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000008611 intercellular interaction Effects 0.000 description 1
- 229940100602 interleukin-5 Drugs 0.000 description 1
- 201000002215 juvenile rheumatoid arthritis Diseases 0.000 description 1
- 239000006249 magnetic particle Substances 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- ZAHQPTJLOCWVPG-UHFFFAOYSA-N mitoxantrone dihydrochloride Chemical compound Cl.Cl.O=C1C2=C(O)C=CC(O)=C2C(=O)C2=C1C(NCCNCCO)=CC=C2NCCNCCO ZAHQPTJLOCWVPG-UHFFFAOYSA-N 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 230000002969 morbid Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000002085 persistent effect Effects 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- REQCZEXYDRLIBE-UHFFFAOYSA-N procainamide Chemical compound CCN(CC)CCNC(=O)C1=CC=C(N)C=C1 REQCZEXYDRLIBE-UHFFFAOYSA-N 0.000 description 1
- 229960000244 procainamide Drugs 0.000 description 1
- 230000006920 protein precipitation Effects 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000001542 size-exclusion chromatography Methods 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000009870 specific binding Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 229960002317 succinimide Drugs 0.000 description 1
- 230000008093 supporting effect Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 229940104230 thymidine Drugs 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000010474 transient expression Effects 0.000 description 1
- 238000002054 transplantation Methods 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 230000029812 viral genome replication Effects 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- -1 α/β TCR Proteins 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2875—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the NGF/TNF superfamily, e.g. CD70, CD95L, CD153, CD154
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/12—Drugs for disorders of the urinary system of the kidneys
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Engineering & Computer Science (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Urology & Nephrology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Epidemiology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Peptides Or Proteins (AREA)
Abstract
A method of treating lupus using anti-gp39 antibodies or fragments is provided. Such treatment has been shown to reverse disease, and in particular lupus-associated kidney disease, the major killer of lupus subjects.
Description
Related application
The present invention relates to anti--receptor of B cell antigen CD40, selectivity is called CD40CR, gp39 or up-to-date CD154 in the document, and the soluble ligand of this receptoroid, comprises containing the proteic fusion molecule of some CD40 at least.This is based on, and is part at least based on can be by combine the activatory discovery that suppresses the B cell that helper T lymphocyte mediates with the protein receptor of new 39kD on the helper T lymphocyte film to the fusion rotein of solubility CD40/ immunoglobulin.The invention provides roughly pure CD40CR receptor; The soluble ligand of CD40CR is provided, comprises anti--gp39 antibody and fragment thereof and contain the proteic fusion molecule of some CD40 at least; And control B is provided the method for cell-stimulating, this method is particularly useful in treatment allergy or autoimmune disease.More precisely, the present invention relates to resist-application of gp39 antibody in the treatment of systemic lupus erythematosus (sle) (SLE) or drug-induced lupus.
Background of invention
The research of Mitchison, Benacerraf and Raff proposes T first
hInteracting with the mutual physical property of B iuntercellular is the basis that humoral immunoresponse(HI) takes place.Subsequently studies confirm that T
hForm physics conjugate (Vitetta etc., Immunol.Rev., 99:193-239 (1987)), B cell response T in these conjugates with II class major histocompatibility complex (MHC) compatibility antigen-offer B cell
h(Barrett etc., J.Immunol., 143:1745-1754 (1989)).Because find T
h-the lymphokine of deriving proposes activated T to the growth and the differentiation performance effective influence of B cell
hNear the soluble factor that discharges mediates the activation of interactional B cell.Yet, without any a kind of lymphokine of molecular cloning, no matter be independent or combination, show the ability that enters the B cell cycle of inducing.Different with soluble factor, activation T
hSerous coat part can induce the B cell cycle to enter (Hodgkin etc., J.Immunol., 145:2025-2034 (1990); Noelle etc., J.Immunol., 146:1118-1124 (1991)).Use activation T
hThe research prompting of purification serous coat part, in activated T
hFilm on a kind of albumen initiating humoral immunization (Noelle etc., J.Immunol., the 146:1118-1124 (1991) that express; Bartlett etc., J.Immunol., 145:3956-3962 (1990)).
To activate T
hPurification serous coat (the PM of cell membrane
ACT) be used to study character (Hodgkin etc., J.Immunol., the 145:2025-2034 (1990) of this effector function; Noelle etc., J.Immunol., 146:1118-1124 (1991)).Activation T
hPM
ACTRather than tranquillization T
h(PM
REST) show the activity of inducing the B cell cycle to enter with in antigen non-specific, the II class-non-limiting way.In addition, confirmation is this by PM
ACTThe activity of expressing need activate 4-6 hour, from new synthetic RNA, and be protein (Bartlett etc., J.Immunol., 1145:3956-3962 (1990)) in itself.
Summary of the invention
The present invention relates to the anti--receptor of B cell antigen CD40 of called after CD40CR and the soluble ligand of this receptor, comprise containing the proteic fusion molecule of some CD40 at least.This is based on, at least be part based on can be by combine the activatory discovery of the B cell that suppresses the helper T lymphocyte mediation with the protein receptor (anti--receptor called after " CD40CR " of CD40) of new 39kD on the helper T lymphocyte film to the fusion rotein of solubility CD40/ immunoglobulin, and the activated discovery that can suppress the B cell that helper T lymphocyte mediates based on the direct monoclonal antibody at this 39kD receptor of called after MR1.
The invention provides CD40CR soluble ligand (comprising antibody) roughly pure CD40CR receptor and contain the proteic fusion molecule of some CD40 at least; And provide control B the method for cell-stimulating.
In specific embodiment of the present invention, contact the activation of the B cell that suppresses the experimenter with experimenter's helper T lymphocyte by treating effective dose soluble ligand or CD40CR.This inhibition to the B cell-stimulating is perhaps particularly useful in the treatment of allergy or autoimmune disease.
More particularly, the present invention is for needing the lupus experimenter of this treatment, as carrying out property systemic lupus erythematosus (sle) or drug induced lupus, even be in the patient of the late stage (this observes kidney damage often) of lysis, Therapeutic Method is provided, this Therapeutic Method is the anti--gp39 antibody for the treatment of effective dose, for example is disclosed in U.S.'s the 08/475th, No. 847 (application on June 7 nineteen ninety-five) of common transfer, present Anti-Human gp39 antibody or its fragment used of having allowed.
An advantage of the invention is that it can get involved an aspect of the immunne response of antigen non-specific.Allergic many therapies comprise the desensitization therapy to specific antigen at present, all need each patient is detected so that identify the antigen relevant with anaphylaxis.Practical problem is, in fact is impossible with the patient to each and every kind of potential allergic effect original work analyzing in detail.And, in most of autoimmune diseasees, even normally unknown the haveing nothing to do of cause of disease antigen with disease process.Relate to the interactional the present invention of antigen non-specific CD40/CD40CR and avoided needs the relevant antigenic characterized of allergy or autoimmunity.Therefore, the present invention's treatment unknown in those immunogens or that have lupus due to the immunogenic allergy of multicomponent and autoimmune disease such as pollinosis, the procainamide or systemic lupus erythematosus (sle) (SLE) has advantage in using especially.Also can be applied to the acute treatment immune activation, as anaphylactoid treatment.
Abbreviation
The Ig immunoglobulin
The Mab monoclonal antibody
PM
ACTSerous coat by activatory helper T lymphocyte preparation
PM
RESTSerous coat by the preparation of tranquillization helper T lymphocyte
The PAGE polyacrylamide gel electrophoresis
RIL4 recombinant interleukin-4
RIL5 recombinant interleukin-5
The SN supernatant
T
hHelper T lymphocyte
Cut line
Fig. 1. monoclonal antibody and CD40-Ig pass through PM
ACTTo the synthetic influence of guiding B cell RNA.
The A group. with tranquillization B cell and T
h1 Pmtest or PM
ACTCultivate together.To containing PM
ACTEach hole in add 25 μ g/ml anti--complex (every kind 25 μ g/ml) of CD4, anti--LFA-1 or anti--ICAM-1 or these materials, and by [
3H]-uridine to mix the RNA that measures the B cell synthetic.The RNA that cultivates assessment B cell after 42-48 hour is synthetic.Given result be three multiple hole cultures arithmetical average+/-standard deviation, and represent 5 such experiments.
The B group. with tranquillization B cell and T
h1 (●, ▲) or T
hThe PM of 2 ()
ACTCultivate together.To containing T
h1PM
ACTThe hole of culture (●, ▲) in add the CD40-IG (▲) or the control protein CD7E-Ig (φ) of increment gradually.To containing culture T
h2PM
ACTAdd the CD40-Ig of increment gradually in the hole ().The RNA that cultivates assessment B cell after 42-48 hour is synthetic.Described result be three multiple hole cultures arithmetical average+/-standard deviation, and represent 3 such experiments.
The C group. with tranquillization B cell and LPS (50 μ g/ml) or PM
ACTCultivate together.In the hole of culture, add CD40-Ig (25 μ g/ml; Hatch) or CD7E-Ig (25 μ g/ml; Solid).It is synthetic to press mensuration RNA described in the A group.Shown in the result be three multiple hole cultures arithmetical average+/-standard deviation, and represent 3 such experiments.
Fig. 2 .CD40-Ig suppresses the differentiation and the propagation of B cell
The A group. with tranquillization B cell and PM
ACT, rIL4 (10ng/ml) and rIL5 (5ng/ml) cultivate together.When cultivating beginning or when cultivating beginning back first, second or the 3rd day, add CD40-Ig or CD7E-Ig (25 μ g/ml).When cultivating the 6th day, collect each hole SN and described, with anti--quantitative IgM of the specific ELISA of isotype (■) and IgG by (J.Immunol., 146:1118-1124 (1991)) such as Noelle
1(●).At PM
ACT, IL4 and IL5 exist (not adding CD40-Ig) down, IgM and IgG
1Concentration be respectively that IgM is 4.6 μ g/ml and IgG
1Be 126ng/ml.When IL4 and IL5 shortage, do not measure IgM or IgG
1The result represents 3 such experiments.
The B group. with anti--CD3 tranquillization or activation T
h116 hours, after the radiation in the presence of IL4 (10ng/ml), with 1 * 10
4/ hole and tranquillization B cell (4 * 10
4/ culture) cultivates together.In culture, add CD40-Ig (▲) or CD7E-Ig (●) from 0-25 μ g/ml.After cultivation 66-72 hour, with 1.0 μ Ci [
3H]-thymidine imposes pulse and collects this culture each hole.Described dotted line represents that the B cell is to tranquillization T
hReplying of cell.Shown in the result be three multiple hole cultures arithmetical average+/-standard deviation, and represent 2 such experiments.
Fig. 3 .CD40-Ig is determined at activation T
hRather than tranquillization T
hThe molecule of last expression.Results tranquillization and activation T
h, and with fusion rotein one arise from 4 ℃ hatch 20 minutes after, add the goat-anti-hIgG (25 μ g/ml) of FITC-coupling.By analyzing at least 5000 cells/sample, measure percentage rate positive cell and MFI.The result represents 6 such experiments.CD40-Ig represents with solid scattergram.
Fig. 4 .CD40-Ig immunoprecipitation activation T
hThe 39kD albumen of 1 solute.With insoluble resisting-CD3 tranquillization or activation T
h116 hours.With antibody purification or fusion rotein (1-10 μ) immunoprecipitation tranquillization or activation T
h[
35S]-albumen of labelling.Described gel scattergram is represented 3 such experiments.
Fig. 5. inductive 39kD T
hThe memebrane protein monoclonal antibody specific suppresses PM
ACTInductive B cell RNA is synthetic.With tranquillization B cell and PM
ACTRespectively with anti--α/β of each 10 μ g/ml anti--CD3, CD40-Ig or MR1 cultivate.It is synthetic to press the described mensuration of Fig. 1 RNA.The result with the arithmetical average of three multiple hole cultures+/-standard deviation represents, and represents 3 such experiments.
Fig. 6 .MR1 and CD40-Ig identification activation T
hThe same molecular of last expression.
The A group. will activate T
hWith MR1 or contrast Ig fluorescence staining.Whether competitive for assessing CD40-Ig and MR1 in conjunction with activation T
h, the MR1 or the contrast hamster Ig (anti--α/β TCR) of variable concentrations gradient joined wherein with anti-CD 40 (20 μ g/ml).After 4 ℃ hatch 20 minutes, the washing sample product and with the human IgG FITC-coupling, anti-
1Monoclonal antibody hatch together.The result represents 3 such experiments.
The B group. usefulness MR1 (10 μ g/ sample) or CD40-Ig (10 μ g/ sample) immunoprecipitation [
35S]-the activation T of methionine-labelling
hAlbumen and separate this albumen with fluorography by PAGE.The result represents 2 such experiments.
Fig. 7 .CD40-Ig combines with the human cell line's.Various human T-cell line is exposed to the CD40-Ig of biotin-labelling, and assesses its combination by the fluidic cell method.
Fig. 8.
The nucleotide sequence (EMBO J., 8:1403-1410 (1989)) of the CD40 cDNA of A group .Stamenkovic etc.Underscore is for striding the film district.
The B group. can be used for expressing the plasmid sketch map of CD40-Ig.The aminoacid sequence of Δ CD40 position of fusion is shown in the proteic diagram part of following CD40.
Fig. 9.
That different treatment group NZB/NZW F1 mices are produced is anti--(ss) titre of dna antibody.The 4-10 monthly age mice that cavity ring (--zero---) representative is treated with MR-1; Hollow triangle (--△---) representative is taking place to accept the mice that MR-1 does not react behind the albuminuria; Closed square (--■---) representative is taking place to accept the mice that MR1 reacts behind the albuminuria, and solid torus (---●---) the representative mice of not receiving treatment.The numerical value that draws is the mean+/-SEM of every group of three titres.
Figure 10.
The survival rate of the NZB/NZW F1 mice of different treatment groups.The 4-10 monthly age mice that cavity ring (--zero---) representative is treated with MR1; Hollow triangle (--△---) representative is taking place to accept the mice that MR1 does not react behind the albuminuria; Closed square (--■---) representative is taking place to accept the mice that MR1 reacts behind the albuminuria, and solid torus (---●---) the representative mice of not receiving treatment.
Detailed Description Of The Invention
The invention provides roughly pure CD40CR acceptor; The soluble ligand of CD40CR is provided, comprises antibody and the fragment and the fusion molecule that contains CD40 of anti--gp39; And provide the using soluble part to control the method for B cell-stimulating.
For openly clear rather than restriction the present invention, detailed description of the present invention is divided into following part:
(i) in conjunction with the part of CD40CR;
(ii) for the identification of the method for CD40CR;
(iii) preparation of the CD40CR of purifying;
(iv) in conjunction with the purposes of the part of CD40CR; With
(v) purposes of CD40CR.
(iii) preparation of purifying CD40CR; (especially treating carrying out property lupus, such as systemic loupus erythematosus or drug-induced lupus).
The invention provides the soluble ligand of CD40CR, comprise (i) contain at least the fusion molecule of some CD40 albumen and (ii) with the antibody of CD40CR specific binding or the CD154 of antibody fragment or gp39 or known described antigen.
Term used herein " solubility " represents that part of the present invention is not always to be combined with the cell serous coat. Yet soluble ligand of the present invention can be fixed on the acellular solid support, comprises lipid, protein or carbohydrate molecule, ball, vesica, magnetic-particle, fiber etc., maybe can be embedded in implant or the vesica.
The ability of such part and CD40CR combination can confirm by the identical protein combination of the described part of proof and CD40-Ig (as follows) or MR1 (as follows) or in conjunction with the another kind of antibody of CD40CR (such as disclosed antibody in No. the 08/475th, 847, U.S. of common transfer).
Part of the present invention can form Pharmaceutical composition with suitable carrier.
The invention provides the solubility fusion molecule for the CD40CR part. This class fusion molecule contains at least a part of CD40 albumen with the second minute sub-connection. The preferred CD40 cross-film district that lacks of described CD40 part. The part of CD40 albumen that can be used according to the invention is defined as any part that can be combined with CD40CR, as showing the part of being combined with same protein such as MR1 or CD40-Ig.
Spendable the second molecule comprises peptide class and protein, lipid and carbohydrate, in the preferred embodiment of the invention, can for immunoglobulin molecules or its part (such as Fv, Fab, F (ab ')2Or Fab ' fragment) or CD8 or other adhesion molecule such as B7. The second molecule can derive from non-human source or human origin, maybe can be chimera. The second molecule also can be a kind of enzyme, toxin, growth factor, lymphokine, antiproliferative, alkylating agent, antimetabolite, antibiotic, vinca alkaloids, platinum coordinate complex, radio isotope or fluorescent chemicals.
Can be by chemical synthesis or preferably produce fusion molecule of the present invention by the DNA recombinant technique.
For example, the nucleotide sequence of a part of CD40 albumen of encoding at least and the nucleotide sequence of second molecule of coding can be made up in suitable expression vector, then in protokaryon or preferred eukaryotic expression system such as yeast, baculoviral or mammalian expression systems (comprising transgenic animals), express.
Perhaps, at least a portion CD40 albumen can or be used with CD40 or second bonded part of molecule and do the affinity chromatograph expression by electrophoretic techniques.Include but is not limited to G28-5 (this hybridoma preserving number be HB9110 and be preserved in American type culture collection) and the CD40CR of anti-CD 40 antibodies with the bonded part of CD40, hereinafter will elaborate as producing by hybridoma.If second molecule is immunoglobulin or immunoglobulin fragment, can use the affinity column that contains anti--immune globulin antibody; If second molecule contains the Fc fragment, can use the protein A post.
According to the preferred embodiments of the invention, the nucleotide sequence of available code CD40 albumen (this albumen blocks from striding the diaphragm area upstream) is produced the part of CD40.Can prepare this nucleotide sequence by the plasmid that digestion contains coding CD40 antigenic cDNA, as it is described press Stamenkovic etc. (EMBO J., 8:1403-1410 (1989)), usefulness
Pst I(P) and
Sau 3A-(S3) Restriction Enzyme digestion.With resulting P/S3 fragment sub-clone go into P and
Bam H I(B) in Xiao Hua the identical plasmid, with the CD40 gene (see figure 8) of production truncate.
In the non-sex-limited specific embodiments of the present invention, being used for producing the expression vector that contains at least a portion CD40 and immunoglobulin sequences can be preferably form from the isolating eukaryotic promoter of DNA sequence and the enhancer sequence of coding constant region for immunoglobulin by viral origin of replication, bacillary origin of replication, bacillary selected marker and by restriction endonuclease sites, after connect the polyadenylic acid signal sequence, described restriction endonuclease sites can allow the DNA sequence (seeing Fig. 8 .b) of sub-clone coding at least a portion CD40.
In particular of the present invention, can be in the fusion plasmid of immunoglobulin with the CD40 gene sub-clone of truncate, as (Cell, 61:1303-1313 (1990)) as described in the Aruffo etc., use
MIu IWith B digestion, make up the pCD40-Ig plasmid, this plasmid-encoded fusion molecule CD40-Ig (see figure 8).Then can by with the pCD40-Ig plasmid transfection to the COS cell, form transient expression system and produce the CD40-Ig fusion rotein.Collect the CD40-Ig that is produced from COS cell conditioned medium liquid, and press Aruffo etc., Cell, 161:1303-1313 (1990) is described to carry out purification through the protein A column chromatography.
Soluble ligand of the present invention preferably includes antibody molecule, monoclonal antibody molecule or contains and the fragment of these antibody molecules of the bonded antigen binding site of CD40CR (gp39) (preferred people gp39).This class part also can contain second molecule, but this molecule protein, lipid, carbohydrate, enzyme, toxin, somatomedin, lymphokine, antiproliferative, alkylating agent, antimetabolite, antibiotic, vinca alkaloids, platinum coordinate complex, radiosiotope or fluorescent chemicals and can be connected with described antibody molecule or fragment.
When described part is monoclonal antibody or its fragment, can use any monoclonal antibody that can provide the technology that produces antibody molecule by the cell line continuous culture to prepare anti-CD40CR (gp39).For example, and the hybridoma technology of at first developing by Kohler and Milstein (1975, Nature, 256:495-497) and recent available other technology, as human B cell hybridoma technology (Kozbar etc., 1983, Immunology Today, 4:72) with the EBV-hybridoma technology (Cole etc. that produce human monoclonal antibodies, 1985, monoclonal antibody and treatment for cancer, Alan R.Liss, Inc., 77-96 page or leaf) all within the scope of the present invention.Usually preferred humanization or mosaic type resist-gp39 antibody, because they are difficult for causing immunogenic response (HAMA replys) after administration.
Can produce the antibody fragment that contains the described molecule of idiotype by known technology.For example, this class fragment is including, but not limited to: F (ab ')
2Fragment, it can produce by using the described antibody molecule of pepsin; Fab ' fragment, it can be by reduction F (ab ')
2Segmental disulfide bond produces; F (ab ')
2Fragment, it can handle described antibody molecule generation by using papain; And 2Fab or Fab fragment, they can be handled described antibody molecule by the Reducing agent of using papain and reduction disulfide bond and produce.
As mentioned above, the present invention also provides by the chimera of technology generation known in the art or people's antibody, as Morrison etc., Proc.Natl.Acad.Sci.U.S.A., technology described in No. the 85305604.2nd, 81:6851-6855 (1984) or european patent application, the announcement No. 0173494 (Morrison etc., announcement on March 5th, 1986).
The immunogen that is used to produce antibody can be any source that contains CD40CR.For example, activated T
h, as can be with activatory people T
hCell is as immunogen.In addition, can use roughly pure CD40CR by the described preparation of following 5.3 parts.If with activated T
hAs immunogen, can test sero-fast anti-activation T
hCell rather than tranquillization T
hThe reactivity of cell.
Described immunogen also can comprise reorganization gp39 or its fragment.In this, clone and express the DNA of coding Mus and people gp39 with recombination method.These protein provide potential immunogen for anti--gp39 production of antibodies.
In a preferred embodiment of the invention, described soluble ligand is an Anti-Human gp39 monoclonal antibody, more preferably humanization or mosaic type Anti-Human gp39 antibody.Following method is used to produce the MR1 monoclonal antibody, and this antibody specificity ground combines with Mus gp39, and can be used to produce other antibody at CD40CR.
With 5-10
6 Activated T
h1 cell (D1.6) is lumbar injection week about, continues 6 all immune hamsters.As anti-Mus T
h1 serum titer is greater than 1: 10, and 000 o'clock, adopt splenocyte and the NSI of immune hamster, carry out cell fusion with Polyethylene Glycol.With flow cytometer tranquillization and the activation T
hFrom the hole of the hybridoma that contains growth, screen supernatant on 1.Further a kind of generation selectivity identification of test activates T
hThe concrete hybridoma of monoclonal antibody, and with its sub-clone to derive MR1.In ascites, produce MR1 also by ion exchange high pressure liquid chromatography (HPLC) purification.
In addition, more preferably according to the U.S. the 08/475th, No. 847 preparation Anti-Human gp39 antibody, document integral body by reference is attached to herein.
The present invention also provides and comprises monoclonal antibody or its segmental part, and this monoclonal antibody or its fragment can competitive inhibition MR1 and the combining of its target antigen or CD40-Ig and its receptor.
Can be by (i) CD40CR in conjunction with CD40, contain the fusion molecule of some CD40 and at least such as the ability of the antibody of MR1; (ii) CD40CR can stimulate the B cell to enter the functional characteristic of cell cycle, propagation and differentiation; And (iii) the cell distribution of CD40CR is identified CD40CR.
Can by CD40CR in conjunction with such as CD40, contain the fusion molecule of CD40 and identify CD40CR at the ability of the part of the antibody of CD40CR.
As hereinafter discussing in more detail, several technology are used to identify CD40CR.For example, confirm that CD40-Ig and MR1 can discern same 39kD molecule.The two all can be from radiolabeled T to find CD40-Ig and MR1
hImmunoprecipitation 39kD protein (Fig. 5 b) in the solute.In addition, the immunoprecipitation of 39kD protein and CD40-Ig has been removed by MR1 from T
hThe antigen of discerning in the solute.
Also available CD40CR stimulates B to enter cell cycle, propagation and differentiation capability evaluation CD40CR.
For example, find activatory (PM
ACT) rather than (PM of tranquillization
REST) T
hCell serous coat (PM) can be induced the RNA of B cell, and synthetic (Fig. 1 a); Activatory this inducing action of indication B cell is not subjected to the influence of antibody as anti--LFA-1, anti--CD4, anti--ICAM-1.Yet,, find that CD40-Ig or MR1 can suppress PM as Fig. 1 b and shown in Figure 6
ACT-inductive B cell activation.
Can by such as [
3H]-uridine mix RNA (as the B cell differentiation, RNA synthetic increase) or by [
3H]-inducing action of the mixing of thymidine (it is synthetic to measure the DNA relevant with cell proliferation) technical measurement B cell activation.For obtaining the optimum determining of CD40CR, can interleukin 4 (IL4) be joined in the culture medium about 10ng/ml concentration the influence of B cell proliferation.
In addition, can be according to the activation of the functional examination B cell of immunoglobulin secretion.For example, CD40CR (roughly pure form, or exist among the PM, otherwise or) can be joined in the tranquillization B cell with IL4 (10ng/ml) and IL5 (5ng/ml).Cultivate after 3 days, can add the culture medium of additional volumes.When the 6th day of cultivating, collect the supernatant (SN) of each culture and measure IgM and Ig according to described (J.Immunol., 146:1118-1124 (1991)) such as Noelle
1Amount.
Also can identify CD40CR by the cell distribution of CD40CR.For example, according to cells were tested by flow cytometry, observe CD40-Ig and activation but not tranquillization T
h1 in conjunction with (Fig. 3).And, observe combining of activating T cell in CD40-Ig and Jurkat cell, HSB2 cell and the human peripheral, but do not show significantly and the combining of cem cell, HPBALL cell or Mus thyoma cell.
For example, be not restriction, can be following be present in CD40CR on the particular cell types (" test cell ") by the flow cytometer evaluation.Available tranquillization (negative control) and activation (positive control) T
hCell parallel testing subject cell.All cells is with about 1 * 10
5The concentration of cell/50 μ l and part (as CD40-Ig or MR1) were cultivated 20 minutes at 4 ℃, added the antibody of the anti--part of FITC-coupling then.Can add final concentration in all samples is iodate third ingot of 2 μ g/ml.For example carrying out flow cytometry analysis on the BD FACSCAN then.After cell forward scattering of forward gate and lateral scattering, red negative (because iodate third ingot exclusion) can determine the green fluorescence logarithm of living cells.
The invention provides roughly pure CD40CR.Can be by following method, by the cell that carries CD40CR such as activatory helper T lymphocyte, Jurkat and this CD40CR of HSB2 cell preparation.
Can be according to described (J.Immunol., 146:1118-1124 (1991)) such as Noelle, by the settlement action of discontinuous sucrose gradient, from suitable cell such as activated T
h1 cell, preparation cell serous coat.Separation of C D40CR by the following method then: take out the diffusion barrier crude extract with gentle detergent, carry out size exclusion chromatography afterwards, affinity chromatograph, immunoprecipitation (as with CD40-Ig or MR1) and/or gel electrophoresis are carried out with being attached to suitable ligand on the solid support (as MR1 or CD40-Ig) in the back again.
Can expect that the proteinic molecular weight that is produced is about 39kD.
The invention provides solubility CD40CR (promptly acellular), this solubility CD40CR can be included in the Pharmaceutical composition that suitable carriers is formed.It further provides and second bonded CD40CR of molecule, and described second molecule can be peptide, protein, lipid, carbohydrate, enzyme, toxin, somatomedin, lymphokine, antiproliferative, alkylating agent, antimetabolite, antibiotic, vinca alkaloids, platinum coordinate complex, radiosiotope or fluorescent chemicals.
The present invention further provides roughly pure CD40CR by chemosynthesis or the preparation of DNA recombinant technique.For example, can be by being inserted into the λ gt10 expression system from the cDNA of activatory helper T lymphocyte preparation, then with MR1 or CD40-Ig in conjunction with screening, identifying the CD40CR-expression cloning, thereby isolate the gene of CD40CR.Perhaps, can with from the cDNA transfection of activatory helper T lymphocyte preparation to the COS cell, with the supernatant of MR1 or CD40-Ig screening transfectional cell, identify the CD40CR product.Can use expression system known in the art then the gene of CD40CR is used to express CD40CR.
The invention provides the method for utilizing with the bonded part control of CD40CR B cell-stimulating.Specifically, it provides the method that suppresses the B cell-stimulating, comprises B cell and T
hThe mixture of cell is exposed to the bonded valid density part with CD40CR.5.1 parts are to the existing explanation of applicable part in front.Can in external or body, implement method of the present invention.Valid density refers to suppress the concentration of the part of B cell-stimulating, and available any technology known in the art (comprising the described technology of aforementioned 5.2 parts) is measured, at least about 30%, and preferred about 75%.Preferred, specific non--restricted embodiment according to the present invention can be with CD40-Ig as part, and valid density can be about 10 μ g/ml at least in the case.In another concrete non--restricted embodiment of the present invention, can use the MR1 monoclonal antibody, valid density can be about 10 μ g/ml at least in this case.If implement described method in vivo, the plasma concentration or the local concentration of the valid density assignment body of part.For example, preferably suppress the activation of B cell, so that restriction is to whole immune influence at regional area.
In specific embodiment, the invention provides curee's the method that treatment suffers from the illness relevant with the B cell-stimulating, comprise give therapeutic dose to the experimenter with the bonded part of CD40CR.The experimenter can not be human or preferred human.
The illness relevant with the B cell-stimulating is including, but not limited to allergy (comprising anaphylaxis); Autoimmune disease comprises drug induced lupus, systemic lupus erythematosus (sle), adult's rheumatoid arthritis, juvenile rheumatoid arthritis, scleroderma, Sjogren Cotard etc.; And the viral disease that relates to the B cell, comprise that Epstein-Barr virus (Epstein-Barr) infects and comprise the retroviral infection of HIV (human immunodeficiency virus) infection.
Owing to proposed the B cell-stimulating and to bring out the incubation period human immunodeficiency virus replication relevant, preferably described part of the present invention given HIV (human immunodeficiency virus) (HIV) positive individuals that those also do not develop into acquired immune deficiency syndrome (AIDS) (AIDS) or ARC.
As mentioned above, and in the following embodiments, especially preferably use anti--gp39 antibody or its fragment, comprise its application in drug induced lupus or systemic lupus erythematosus (sle).As the master data of following embodiment and confirmations of experiment institute, found that the treatment of described resisting-gp39 antibody has reduced the generation of autoantibody and alleviated nephropathy, and made the prolongation of NZB/NZW (animal model of generally acknowledged human SLE) survival period.
In addition, in the mice (activeness lupus state) of albuminuria 2-3+, further confirm, resist-gp39 antibody reverse disease (evidence is to give behind the antibody to prolong life cycle and not have albuminuria) unexpectedly.Therefore, after the animal model generation nephropathy of people's lupoid of generally acknowledging, show, reverse the lupus process with anti--gp39 Antybody therapy.This confirms that in treatment lupus and other autoimmune disease, anti--gp39 antibody is used for the probability of human therapeutic use.Particularly it has confirmed the patient that this antibody-like can be used for the treatment of the activeness PD even be in stage terminal stage of a disease.Disease process such as the common renal damage of lupus patient will be treated even may be reversed to this treatment effectively.
In suitable pharmaceutical carrier, can pass through any method afford part known in the art, comprise intravenous injection, lumbar injection, subcutaneous injection, intrathecal injection, intra-articular injection or intramuscular injection, and oral cavity, intranasal, ophthalmic and rectally, but and in its microsphere, liposome and/or the slow release implant.
The therapeutic dose of part is defined as the amount of the clinical adverse effect that can significantly alleviate B cell-stimulating or T cell activation, and can with used part and treat and change.If use CD40-Ig, no matter be that system's (plasma concentration) or topical therapeutic concentration can be about 10 μ g/ml then.If use MR1 or other anti--gp39 antibody,, no matter be that system's (plasma concentration) or topical therapeutic concentration can be about 10 μ g/ml then as humanization or mosaic type Anti-Human gp39 antibody or its fragment.
In further embodiment of the present invention, said method can utilize the part that contains toxin or antimetabolite, will kill T like this
hCell or make it impaired is because destroy T
hCell makes increases the B cell-stimulating.
Also available part of the present invention comes labelling activated T cell, is a kind of technology of diagnosis of the T of can be used for cell disease.For this reason, can be with the part that contains enzyme, radiosiotope, fluorescent chemicals or other detectable label be exposed to the T cell in the external or body, measure described binding capacity.
Also part of the present invention can be used to transmit the material such as the somatomedin of activating T cell.
The invention provides the method for the molecular Control B cell-stimulating that utilizes CD40CR or contain CD40CR, described CD40CR or the molecule that contains CD40CR are to prepare according to preceding method.Specifically, provide the method that promotes the B cell-stimulating, comprise the CD40CR of B cellular exposure in valid density.Can be in vivo or this method of external enforcement.Valid density refers to induce the concentration of the receptor of B cell-stimulating, can pass through any technical measurement known in the art, at least about 30%.The present invention specific, in the non-limiting embodiments, the CD40CR concentration of part or system can be about 10 μ g/ml.
In specific embodiment, the present invention provides Therapeutic Method for the experimenter suffer from humoral immunization weakens relevant immune deficiency disorder, comprises the CD40CR that gives experimenter's therapeutic dose.The experimenter can not be human, but preferred human.
Weaken relevant immune deficiency disorder with humoral immunization and comprise acquired immunodeficiency, as causing by chemotherapy or radiotherapy, and the hereditary that relates to humoral immunization.
In suitable pharmaceutical carrier, can pass through any method afford CD40CR known in the art, comprise intravenous injection, lumbar injection, subcutaneous injection, intrathecal injection, intra-articular injection or intramuscular injection, and oral cavity, intranasal, ophthalmic and rectally, and it can be included in microsphere, liposome and/or the slow release implant.
The CD40CR therapeutic dose of CD40 is defined as can increase the amount that produces about 30% immunoglobulin at least.
In further embodiment, CD40CR can be coupled on the toxin, be in preferred destruction and express experimenter under the situation of B cell of CD40.The example of this situation comprises to be accepted organ transplantation or suffers from multiple myeloma or the patient of other B cell malignancies or autoimmune disease.
Also CD40CR can be used for the B cell of marker expression CD40, be a kind of technology that can be used for the diagnosis of B cell disease.For this reason, will be with enzyme, radiosiotope, fluorescent chemicals or the bonded receptor of other detectable label be exposed to the B cell in the external or body, and measure described bonded amount.
Also available CD40CR passes to the B cell with molecule bonded with it.
New receptor CD40CR on the activatory helper T lymphocyte is thin in conjunction with the CD40 and the B that transduces
Born of the same parents' relatedness activation signal
Material and method
Animal
(Jackson Laboratories, Bar Harbor ME) are used to prepare filler cell (filler cell) to support T with female DBA/2J mice
hClone's growth, and be used to prepare tranquillization B cell.
D 1.6 is a kind of I-A
d-restrictive rabbit immunoglobulin-specificity T
h1 clone (Kurt-Jones etc., J.Exp.Med., 166:1774-1787 (1987)), derive from David doctor Parker (University of Massachusetts, Worcester).D 1.6 is referred to herein as T
h1.
Described according to (J.Immuno., 146:1118-1124 (1991)) such as Noelle, with T
h1 (8 * 10
6/ hole) (6 orifice plates, Corning cultivate in NY), and wrap by 16 hours with anti--CD340 μ g/4ml PBS in each hole of this plate in a bunch collection hole.
Described according to (the same) such as Noelle, prepare the cell serous coat by the discontinuous sucrose gradient sedimentation.
Described according to (J.Exp.Med., 155:1523 (1982)) such as Defranco, by preparing tranquillization spleen B cell in discontinuous Percoll (Percoll) gradient sedimentation.From the cell of the interfacial separation of 70-75% (density is 1.087-1.097) Percoll usually＞95%mIg
+, have evenly, lowly do not reply near the light scattering of forward direction and to Con A.
With ion exchange HPLC following monoclonal antibody of purification from mice (this mice was rebuild already through overshoot and bone marrow) ascites: anti--CD3:145-2C11 (Leo etc., Proc.Natl.Acad.Sci.USA, 84:1374-1378 (1987)); Anti--α, β: H57-597; Anti--CD4:GK1.5 (Wilde etc., J.Immunol., 131:2178-2183 (1983)); Anti--ICAM:YN1/1.7.4 (Prieto etc., Eur.J.Immunol., 19:1551-1557 (1989)); Anti--LFA-1:FD441.8 (Sarmiento etc., Immunol.Rev., 68:135 (1982)); And anti--rat/hamster κ chain: RG-7 (Springer, Hybrid., 1:257-273 (1982)).
By using Restriction Enzyme
Pst I(P) and
Sau3A (S3) digested plasmid prepares the fusion rotein of CD40, and this plasmid contains the coding antigenic cDNA of CD40 (Stamenkovic Seed, EMBO J., 8:1403-1410 (1989)).With this P/S3 fragment sub-clone in same plasmid, this plasmid with P and
Bam H1(B) digestion.The proteinic CD40 Δ of CD40 that allows the preparation coding to block from the membrane spaning domain upstream.To encode the then dna fragmentation sub-clone (Aruffo etc., Cell, 61:1303-1313 (1990)) in this immunoglobulin fusion plasmid of CD40 Δ is used
MIu IDigest with B.According to described (Cell, 61:1303-1313 (1990)) such as Aruffo, produce described CD40-Ig fusion rotein in the COS cell and with its purification on the protein A post by transient transfection.
Interleukin-4 (IL4): reorganization Mus IL4 by C.Maliszewski and doctor's K.Grabstein generosity provide (Immunex Corporation, Seattle, WA.).
T cell growth factor (IL5): reorganization Mus IL5 is available from R﹠amp; The D institute (Sarrento, CA.).
With 3 * 10
4(Costar, Cambridge are incubated among the 50 μ l cRPMI in MA) tranquillization B cell in the A/2 micro titer plate well.In these holes, add 0.5 μ g T
h1 or T
h2 memebrane proteins.At 42-48 hour, each hole was with 2.5 μ Ci
3(New England Nuclear, Boston MA) imposes pulse to the H-uridine, collects each hole, and measures radioactivity by the liquid scintillation spectroscopy.Described result with the cpm/ culture+/-standard deviation represents.
Cultivate tranquillization B cell as mentioned above.In culture hole, add 0.5 μ g T
h1 memebrane protein, IL4 (10ng/ml) and IL5 (5ng/ml).Cultivated the 3rd day, and added 50 other μ l cRPMI.Cultivated the 6th day, and collected the supernatant in each hole and described, measure IgM and IgG by (J.Immunol., 146:1118-1124 (1991)) such as Noelle
1Amount.
With 4 * 10
4(Costar, Cambridge are incubated among the 50 μ l cRPMI in MA) tranquillization B cell in the A/2 micro titer plate well.In these holes, add 1 * 10
4Tranquillization or activation, radiating (500 rads (rad)) T
h1 and IL4 (10ng/ml).Cultivated the 3rd day, according to Noelle etc. at J.Immunol.,
146: described in the 1118-1124 (1991), each hole is with 1 μ Ci
3H-thymidine burst process.
With 5-10 * 10
4Activated T
h1 (D1.6) be lumbar injection every other week, continues immune 6 weeks of hamster.As anti-Mus T
h1 serum titer is greater than 1: 10, and 000 o'clock, adopt splenocyte and the NSI of immune hamster, carry out cell fusion with Polyethylene Glycol.With flow cytometer tranquillization and the activation T
hScreen the supernatant in the hole of the hybridoma that contains growth on 1.Further test is a kind of can produce selectivity identification activation T
hThe specific cross tumor of monoclonal antibody, and with its sub-clone with the MR1 that derives.In ascites, produce MR1 also by ion exchange HPLC purification.
Collect tranquillization and activation T
h(with anti--CD3 16 hours) and with 1 * 10
5Cell/50 μ l and fusion rotein were cultivated 20 minutes at 4 ℃, added goat-anti-people (h) IgG (25 μ g/ml of FITC-coupling then; Southern Biotechnology, Birmingham, AL).Adding iodate third ingot to final concentration in all samples is 2 μ g/ml.On BD FACSCAN, carry out flow cytometry.After cell forward scattering of forward gate and lateral scattering, red negative (because iodate third ingot exclusion) can determine the green fluorescence logarithm of living cells.
The mensuration of positive percentage and MFI will be analyzed 5,000 survivaling cells at least.Utilize the RG7 of FITC-coupling with MR1 dyeing, be the monoclonal antibody of mouse anti-rat/hamster κ chain.
With insoluble resisting-CD3 tranquillization or activation T
h1 16 hours.Use 1mCi[
35S]-methionine/cysteine labelling tranquillization and activation T
h(20 * 10
6/ ml) protein 1 hour, this moment as described (Noelle etc., (1986) J.Immunol.,
137: 1718-1726), its usefulness is contained the RPMI washed twice of 10% hyclone and the described cell precipitation of dissolving in extraction buffer.Solute to 500 μ l (is equivalent to 5 * 10
6Cell) adds antibody purified or fusion rotein (1-10 μ g) in 4 ℃ of effects 16 hours.At this moment, solute is moved in the test tube that contains 50 μ l filling protein A-agarose.Protein precipitation A-agarose is suspended again and test tube was hatched 1 hour in 4 ℃ of vibrations.Use 3 * high stringency lavation buffer solution washing sample then.Sedimentary protein A-agarose is suspended in again in the SDS sample buffer of 30 μ l, and carries out electrophoresis in 10% polyacrylamide gel.Behind running gel, fixing described gel also carries out fluorography.
For determining described cell surface molecule, this is numerator mediated by PM
ACTInduce to enter the B cell cycle, to PM
ACTWith add at T in the culture of B cell
hThe monoclonal antibody of memebrane protein.PM
ACTInducing the RNA of B cell synthetic is to use PM
REST(Fig. 1 a) for 8 times of observation.Add anti--LFA-1, anti--CD4, anti--ICAM-1 separately or with combining form, all can not suppress by PM
ACTThe B cell RNA that causes is synthetic.
In human system, confirmed that the monoclonal antibody of anti-CD 40 is induced B cell proliferation (Clark and Lane, (1991) Ann.Rev.Immunol.9:97-127), therefore hint that CD40 is the important triggering molecule of B cell.Whether participate in for measuring CD40 by PM
ACTSynthesizing of the B cell RNA that causes is with extracellular domain and the human IgG of people CD40
1The soluble fusion protein of Fc domain (CD40-Ig) joins PM
ACTIn the culture of B cell.Preparation is from T
h1 and T
h2 deutero-PM
ACTAnd use it for and stimulate the synthetic of B cell RNA.CD40-Ig adding culture causes the inhibition to the synthetic dose dependent of B cell RNA, by T
h1 and T
h2 deutero-PM
ACTCause, and CD40-Ig is about 5 μ g/ml.And CD7E-Ig fusion rotein (Damle and Aruffo, Proc.Natl.Acad.Sci.USA
88: 6403-6407 (1991)) even when using 25 μ g/ml, still do not have an effect.
For whether research CD40-Ig suppresses the activation of the B cell that caused by T-dependent/non-dependent activator, cultivation B cell in the presence of LPS and the CD40-Ig is being arranged.Second day, evaluation RNA synthetic (Fig. 1 c).The B cell-stimulating unrestraint effect of CD40-Ig to causing by LPS, but suppress the B cell to PM
ACTReply.
At PM
ACT, when IL4 and IL5 exist, the differentiation of B cell polyclone produces Ig (Hodgkin etc., (1990) J.Immunol.
145: 2025-2034; Noelle etc., (1991) J.Immunol.
146: 1118-1124).For estimating in this process needs, cultivating initial or adding CD40-Ig in a couple of days subsequently of cultivating to the CD40 signal.(control level of Fig. 2 during a) with its shortage compared, and it is to producing polyclone IgM and IgG cultivating the adding of initial CD40-Ig
1Inhibition greater than 95%.In contrast, show then that in first day of cultivation and the adding of second day CD40-Ig it is to IgM and IgG
1Generation inhibitory action if any, also be atomic little.After these data showed 24 hours, the signal that sends by CD40 no longer was essential for the differentiation secretion Ig of B cell.
Therefore, data have further pointed out CD40 to participate in PM
ACTThe activation of the B cell that causes.For confirming that CD40 is also with by complete, survival, activated T
hThe activation of the B cell that causes is relevant, studies.Activate T with insoluble resisting-CD3
h1 16 hours, collect and the described T of radiation
h1.In the presence of IL4, with the T of this radiation
h1 cultivates and is cultivating the propagation that detected the B cell on the 3rd day with the B cell.Use T
h1 needs ectogenic IL4 when reaching B cell proliferation, because T
h1 does not produce IL4 (Noelle etc., (1989) J.Immunol.
143: 1807-1814).With use PM
ACTObservation identical, CD40-Ig suppresses T by radiation in the dose dependent mode
hThe propagation of the B cell that causes (Fig. 2 b).Its negative control, CD7E-Ig do not show obvious effect.
For observing activated T
hWhether 1 express conjugated protein to CD40, with tranquillization and activated T
h1 (16 hours) resist-HigG with FITC-subsequently with CD40-Ig or CD7E-Ig dyeing.Combination (Fig. 3) by flow cytometer method assessment CD40-Ig.To activate 16 hours T with anti--CD3
h1, but not tranquillization T
h1, the CD7E-Ig dyeing that need not contrast, positive rate 56% with CD40-Ig.Conjugated protein for identifying this CD40-Ig, with T
h1 protein usefulness [
35S]-methionine/cysteine biosynthesis labelling and with CD40-Ig or CD7E-Ig immunoprecipitation protein.The protein of this immunoprecipitation is separated and fluorography (Fig. 4) with SDS-PAGE.Article one, significantly the apparent molecular weight of band is that 39kD and a low-molecular-weight band are β2Wei Qiudanbais.When lacking monoclonal antibody, then can't see significant band.Also can be from using
125The activation T of I vector (vectorially) labelling
hMiddle immunoprecipitation 39kD band confirms that described 39kD protein is memebrane protein.
Developed at activating T
hGo up and not at tranquillization T
hLast selective expression's antigenic monoclonal antibody specific causes T in order to evaluation
hActive T of effective stage
hMolecule.A kind of such monoclonal antibody MR1 is identified in activation T
hSelective expression's antigen on 1.Whether MR1 discerns identical molecule with CD40-Ig for research, has carried out fluidic cell and has learned and block research.The T of activatory rather than tranquillization
hCell, the dyeing of its CD40-Ig and MR1 are approximately 56% and 61% respectively, and (Fig. 5 a).MR1, but be not that another hamster resists-the anti-α of T cell monoclonal antibody/β TCR, block activation T in the dose dependent mode
hThe dyeing of cell and CD40-Ig.These Notes of Key Datas CD40-Ig and MR1 identification 39kDT
hOverlapping or same epi-position on the albumen.Be further to confirm the identical molecule of CD40-Ig and MR1 identification, by from radiolabeled T
hImmunoprecipitation protein in the solute is identified the described antigen in conjunction with MR1.The protein of the equal immunoprecipitation 39kD of CD40-Ig and MR1 (Fig. 1 .5b).At last, the immunoprecipitation of 39kD protein and CD40-Ig has been removed origin self-activation T
hRadiolabeled solute in the antigen of MR1 identification, support that described MR1 antigen and CD40 conjugated protein are this identical principles.
Carry out functional study with MR1, whether neutralize by PM to illustrate this monoclonal antibody
ACTThe activity of expressing.With PM
ACTCultivate separately or in the presence of hamster monoclonal antibody or CD40-Ig with the B cell.Two kinds of hamster monoclonal antibodies, anti-α/β TCR and α-CD3 can not suppress by PM
ACTThe activation of the tranquillization B cell that causes.In contrast, MR1 or CD40-Ig suppress B cell-stimulating (Fig. 6).
The T that described data show is given prominence to the monoclonal antibody blocking-up
hSurface molecular (LFA-1, CD4, ICAM-1, CD3, α/β TCR) does not hinder activation T
hInduce the ability that enters the B cell cycle.In contrast, specificity is blocked the activation of T-dependency B cell at protein-bonded CD40-Ig of CD40 or monoclonal antibody in the dose dependent mode.And conjugated protein being accredited as of described CD40 is in activation but not the T of tranquillization
hThe protein of selective expression's 39kD on the film.The two all blocks by PM specificity at the protein-bonded CD40-Ig of described 39kD CD40 and monoclonal antibody
ACTThe activation of the B cell that causes.
Although many memebrane proteins and T
hIt is relevant that-dependency B cell sends signal, but evidence provided herein has been got rid of the effect of some molecule (LFA-1, CD4, CD3, α/β TCR, ICAM-1), and hint CD40 is used for by T as B-cell receptor
hThe related signal that sends.The data show specificity suppresses T at protein-bonded CD40-Ig of CD40 and monoclonal antibody
hThe activation of-dependency B cell.
The part of CD40 is to be expressed in T activatory but not tranquillization
hOn the protein of 39kD.Biochemical Research points out that this 39kD protein is a single chain molecule, because electrophoretic migration is not subjected to the influence of Reducing agent.According to the functional study that in this research, provides, activated T
h1 and T
h2 the two all to express described 39kD CD40 conjugated protein.This is consistent with described functional study, the described T that studies show that
h1 and T
h2 the two all induce B to enter cell cycle.When attempting further to identify the proteinic feature of described 39kD, be that the cDNA transient transfection of CD protein (Cd53, CD27 and CD69) of 39kD is in the COS cell and test the combination of the CD40-Ig of described cell with the coding molecule weight range.The neither one transfectional cell is expressed the protein in conjunction with CD40-Ig.Therefore guess that this 39kD protein is not a kind of in this class CD protein.
At T
hWith the biochemical basis of B iuntercellular conducted signal be to make us unintelligible.Discriminating to as the CD40 of the molecule of helper T lymphocyte conducted signal causes the concern of known coupling in the specificity biochemical route of CD40 molecule.By means of 4 existence of being rich in the primitive of cysteine in its extracellular region territory, CD40 becomes a member of trk C (NGFR) family.Shown that monoclonal antibody sends signal (Uckun etc. by CD40, J.Biol.Chem.266:17478-17485 (1991)) participate in the activation of tyrosine kinase, the result has increased the activation of the serine/threonine kinase of the generation of inositoltriphosphoric acid and at least 4 uniquenesses.Based on sending the information that obtains the signal from described other member of NGF receptor family, expection activation T
hWith the intercellular interaction of B will generation effect in the many links of same biochemical process.
In conjunction with research, use biotin-butanimide (Sigma) about immunofluorescence with CD40 Ig fusion rotein and biotin coupling.Carrying out the fluidic cell credit by two step stainings and Coulter Epics C instruments with phycoerythrin (PE)-Streptavidin (Becton-Dickinson) then analyses.The representative result who screens a plurality of T cell lines offers below.Find Jurkat and the combination specifically of HSB2 cell line, other T cell line comprises that CEM, HPBALL and mice thyoma then do not combine (Fig. 7) with CD40 Ig fusion rotein.
Be used for the treatment of or prevent the application of the anti-gp39 antibody of lupus
Systemic lupus erythematosus (sle) (SLE) is a kind of disease that produces multiple pathogenic autoantibody (Boumpas, Ann Int Med, 1995) that is characterized as.These antibody directly by identification on the normal cell antigenic determinant or indirectly the formation by immune complex cause infringement, described immune complex can be deposited in the normal structure and the activating complement cascade system.
As normal antibody response, from research, check at present the SLE of inbrde mouse species and class-lupus, the generation of lupus B cell autoantibody depends on CD4
+Complementary T (T
h) synergism of cell.Embodiment comes from the research of traditional SLE mouse model, and this model is the female descendant disease Mus of the NZB/NZW that (to comprise the generation of autoantibody and the generation of ICG) in many aspects similar to people SLE.Also reversed nephritis really with the Therapeutic Method prevention that reduces anti-CD 4 antibodies to mice.Unfortunately, the angle from treatment to the transience therapeutic test of mankind itself's immune disease or even anti-CD 4 antibodies, also can cause the long-term depletion of the lymphocytic subgroup that this is important in some case.And anti-CD 4 antibodies is disappointing such as the effect in the small-scale clinical trial in the disease of rheumatoid arthritis.
More specific immunosuppressant is practicable, because identified participation T
hAuxiliary several interactions of molecules of cell, feasible only targeting provide and play an active part in the Th cell of offering help for the antibody generation.An example is to pass through T
hSecondary signal is transmitted in interaction between the B7.1/B7.2 on CD28/CTLA4 on the cell and the B cell.Any one participant's of the two blocking antibody can hang down at the external T of making cell and reply at this.When giving NZB/NZW mice even terminal stage of a disease during sick Mus, the fusion rotein that contains mouse cell external structure territory CTLA-4 and mice Ig Cg2a chain combination at present demonstrates the generation of autoantibody capable of blocking, is to play a role with combining of endogenous CTLA-4 by competitive inhibition B7.2 in the treatment mice by inference.
Experimental result
We have studied in the NZB/NZW Mus and have given the effect of anti-CD 40 L antibody to 4 to 10 monthly age SLE mices.The treatment of anti-CD 40 L antibody has significantly reduced the generation of the Anti-DNA autoantibody of these mices, has alleviated nephropathy and has prolonged life cycle, does not cause that anti-antibody is replied in the mice or causes the infringement of tangible immunity system easily in treatment.These packets are contained among Fig. 6.When 11 monthly ages, the mice of 60% anti-CD 40 L Antybody therapy still survives, rather than the mice (Fig. 9) of treatment or Hig treatment not.The inspection of the nephridial tissue of long-term survivors behind the antagonism-CD40L Antybody therapy shows inappreciable pathological change and does not have significantly immunity deposition at glomerule.
The data that obtain are also pointed out, anti-CD 40 L antibody therapy can avoid two main uses anti--the immunotherapy misgivings of the antibody of lymphocytic cell surface molecule: anti-antibody takes place reply and treat the back permanent immunity and suppress.In our one group of experiment (cohort), antagonism-CD40L Antybody therapy effectively, the mice of survival anti-antibody does not take place is replied in the time of 11 months, almost is a fixing phenomenon in the animal of accepting another kind source antibody.We infer that this is because anti-CD 40 L antibody has prevented the production of antibodies at self.In the normal mouse research of handling with identical anti-CD 40 L antibody MR-1, do not observe replying of anti--MR-1, point out us observed replying on the NZB/NZW mice and may represent a kind of special allergy that is confined to the autoimmunity mice.In a word, this observes for the therapeutic application of Anti-Human CD40L antibody significant, because particularly use " humanization " type Antybody therapy to avoid the severe complication of Antybody therapy with anti-CD 40 L antibody.In our research, the mice that anti--anti-CD 40 L antibody response takes place is good unlike control mice aspect its Anti-DNA production of antibodies or existence, in any case prompting, anti-CD 40 L antibody can not prevent that the generation antibody of such mice is anti-.Anti--anti-CD 40 L antibody that anti-CD 40 L production of antibodies can cause quick removing to give subsequently, and therefore lose the treatment curative effect.For supporting this viewpoint, when handling mice, identify in the glomerule of the reactionless mice subgroup of anti-CD 40 L antibody treatment and find hamster antibody deposition that anti-CD 40 L antibody responder does not then have this phenomenon with non-specific hamster IgG.This observation with anti--formation and the deposition of anti-CD 40 L antibody mediated immunity complex in the kidney that gives anti-CD 40 L antibody continuously is consistent, this may make glomerulonephritis worsen but not prevention.
At last, confirmation is along with stopping with anti-CD 40 L Antybody therapy, anti-after 4 months-KLH antibody response ability is recovered at the most, and establishing described antibody is potential reversible to the immunosuppressive action of immunity system, so the therapy of anti-CD 40 L antibody is applicable to human treatment's application.
In these experiments, with the MR-1 of 200 μ g weekly twice lumbar injection give 4-10 monthly age NZB/NZW mice.Equally, by MR-1 dosage being increased to 250 μ g, on every Wendesdays time, 4 to 10 monthly age of lumbar injection mice therapeutic scheme, realize treatment.This dosage is used for the rheumatoid arthritis model of collagen protein-cause, causes treating mice 100% survival (Durie, Science, 1993).Use this scheme, present described NZB/NZW mice is all 100% survivals (Figure 10) when 11 monthly ages, and two matched groups of mice are all dead when 10 monthly ages.
Determine the lupus treatment of diseases
Although purpose is to prevent the SLE of NZB/NXW F1 mice with anti--gp39 antibody, indication can not be used to this antibody to get involved activeness progressivity morbid state.Whether the available antibodies treatment reverses the disease of being established in order to measure, and one group of mice is developed into urine dipstick mensuration 2-3+ albuminuria (suitable and 100mg/ days to 500mg/ days).At this moment, the mice random packet is not treated group or accepted MR1 (100 μ g/ days) group for continuing.Do not treat mice all death when 10 monthly ages for described 10.In contrast, 5 survival to 11 monthly ages in 10 MR-1 treatment mice.Wherein 3 mices are healthy and do not have an albuminuria.Yet, what is interesting is that 1 Anti-DNA antibody (table 1) that height-titre do not occur is arranged in these mices.In 5 survival Mus 2 Anti-DNA antibody and 4+ albuminuria that show illness and the high titre of tool are arranged.
Therefore, find that in nephropathy the back taking place treats the disease that mice is divided in reversible transfer part with MR1.Reply in the mice at 1 and to find to have reversed albuminuria, but do not reverse the Anti-DNA production of antibodies, further increased MR1 and brought into play and be different from the probability of eliminating other mechanism that T-dependency antibody produces with the MR1 treatment.
Equally, also show development (Jacob, Science, 1988 that can significantly delay nephritis with tumor necrosis factor (TNF-α) treatment NZB/NZW mice; Gordon, 1989).Interesting is, observed heavy nephritis in NZB/NZW F1 mice, part are owing to be positioned dominant gene on the H-2 complex from what the NZW parental generation obtained.Described NZW mice has the TNF-alpha levels of reduction, and this level is relevant with the polymorphism of the TNF-α gene that is arranged in the H-2 complex.Described NZB/NZW F1 mice also has the level of significantly reduced TNF-α.In addition, the NZB/NZW F1 mice with anti--IL-10 Antybody therapy has postponed the generation of nephritis basically and has prolonged survival period.When with anti-TNF-Alpha antibodies therapeutic alliance, the beneficial effect of mediation appears being raised by the inductive TNF-alpha levels of IL-10-in the treatment mice, and eliminated anti--IL-10 and replied.
In preliminary experiment, we have also measured the level of mice TNF-α with ELISA, this mice be respectively give before and give the back MR-1 4 treatment groups.Described level is shown in table 2.According to the reduction of albuminuria and the generation of Anti-DNA antibody, only there is pair mice that the MR-1 treatment is replied to confirm that TNF-α is from treating the 35.01pg/ml after preceding 2.16pg/ml is increased to treatment.The described MR-1 effect of these Notes of Key Datas can partly be passed through the increase mediation of the TNF-α of NZB/NZW F1 mice at least.
In addition, all to confirm to utilize anti--gp39 antibody and its active fragment to be used for the treatment of human lupus be systemic lupus erythematosus (sle) or drug-induced lupus to these results.Particularly, these antibody can be used for the activeness that feature often shows as nephritis, the treatment of carrying out property lupus gets involved.This antibody-like should suppress the process of described disease, even the probability that reverses the course of disease is arranged.
Table 1
After age MR1 treatment treatment back alleviation was treated with the MR1 persistent period
(moon) (week) albuminuria (week) is treated (week) albuminuria again
7 13 +4 N/A N/A - -
9600 nothings--
660 12 nothings--
560 11 have 6 (0) 0
81 death----
6 18 death----
6609 have 3 (+4)+4
66 death----
75 death---
62 death---
Table 2.
| Mice * | The age (mos.) of institute's sample thief | TNF-α(pg/ml)# |
| Not | 2 | 5.5 |
| 9 | 3.1 | |
| The MR-1 | 2 | 2.2 |
| 9 | 35.0 | |
| MR-1 | 2 | 0.0 |
| 8 | 4.3 |
*1 mice of each treatment group.
# value representation TNF-α ELISA test kit (Genzyme, Cambridge, MA) meansigma methods of the duplication of Ce Dinging.
The multiple public publication of this paper citation integral body by reference is attached to herein.
Claims (10)
1. at least a specificity of effective dose is used for making the purposes of suffering from the medicine that the albuminuria that shows as albuminuretic lupus patient alleviates in conjunction with the antibody of CD40L or its fragment in production.
2. the purposes of claim 1, wherein said medicine also are used to make nephritis to alleviate.
3. the purposes of claim 2, wherein said medicine also is used to reverse nephropathy.
4. the purposes of claim 1, wherein said antibody or its segmental dosage range are 10 μ g/ml-1000 μ g/ml.
5. the purposes of claim 1, wherein said patient is in the lupus late stage that shows as kidney damage.
6. the purposes of claim 1, wherein said antibody or its fragment are in conjunction with human CD 40 L.
7. the purposes of claim 1, wherein said medicine and solubility CD40 or solubility CD40 fusion rotein give jointly.
8. the purposes of claim 1, wherein said antibody is anti-human CD 40 L antibody.
9. the purposes of claim 1, wherein said medicine is used to alleviate or alleviate the nephropathy relevant with lupus, but can not block the generation of anti-DNA antibody.
10. the purposes of claim 1, wherein said medicine are used to make TNF-α to produce to be increased.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US5448898A | 1998-04-03 | 1998-04-03 | |
| US09/054,488 | 1998-04-03 |
Related Child Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200410078988XA Division CN100352497C (en) | 1998-04-03 | 1999-04-02 | Antibody inhibition body fluid immunity of using special integrated CD40CR(CD40 part) |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1303301A CN1303301A (en) | 2001-07-11 |
| CN1173735C true CN1173735C (en) | 2004-11-03 |
Family
ID=21991433
Family Applications (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB998067563A Expired - Fee Related CN1173735C (en) | 1998-04-03 | 1999-04-02 | Use of anti-gp 39 antibodies for treatment and/or reversal of lupus and associated kidney disease |
| CNB200410078988XA Expired - Fee Related CN100352497C (en) | 1998-04-03 | 1999-04-02 | Antibody inhibition body fluid immunity of using special integrated CD40CR(CD40 part) |
Family Applications After (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB200410078988XA Expired - Fee Related CN100352497C (en) | 1998-04-03 | 1999-04-02 | Antibody inhibition body fluid immunity of using special integrated CD40CR(CD40 part) |
Country Status (9)
| Country | Link |
|---|---|
| EP (1) | EP1067954A4 (en) |
| JP (1) | JP2002510643A (en) |
| KR (1) | KR20010072564A (en) |
| CN (2) | CN1173735C (en) |
| AU (1) | AU743824B2 (en) |
| HU (1) | HUP0101689A3 (en) |
| NO (1) | NO20004966L (en) |
| TW (1) | TWI224969B (en) |
| WO (1) | WO1999051258A1 (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| MA41459A (en) | 2015-02-03 | 2017-12-12 | Als Therapy Development Inst | ANTI-CD40L ANTIBODIES AND METHODS FOR TREATING CD40L ILLNESSES OR DISORDERS |
| US10669343B2 (en) | 2015-08-05 | 2020-06-02 | Janssen Biotech, Inc. | Anti-CD154 antibodies and methods of using them |
| GB2578037A (en) | 2017-05-24 | 2020-04-15 | Als Therapy Development Inst | Therapeutic anti-CD40 ligand antibodies |
Family Cites Families (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5672347A (en) * | 1984-07-05 | 1997-09-30 | Genentech, Inc. | Tumor necrosis factor antagonists and their use |
| US5474771A (en) * | 1991-11-15 | 1995-12-12 | The Trustees Of Columbia University In The City Of New York | Murine monoclonal antibody (5c8) recognizes a human glycoprotein on the surface of T-lymphocytes, compositions containing same |
| IL104684A0 (en) * | 1992-02-14 | 1993-06-10 | Bristol Myers Squibb Co | The cd40cr receptor and ligands therefor |
| US5876950A (en) * | 1995-01-26 | 1999-03-02 | Bristol-Myers Squibb Company | Monoclonal antibodies specific for different epitopes of human GP39 and methods for their use in diagnosis and therapy |
| US5833987A (en) * | 1995-06-07 | 1998-11-10 | Trustees Of Dartmouth College | Treatment of T cell mediated autoimmune disorders |
| US6001358A (en) * | 1995-11-07 | 1999-12-14 | Idec Pharmaceuticals Corporation | Humanized antibodies to human gp39, compositions containing thereof |
| EP0948355A1 (en) * | 1997-01-10 | 1999-10-13 | Biogen, Inc. | Treatment of lupus nephritis with anti-cd40l compounds |
| WO1998039026A2 (en) * | 1997-03-07 | 1998-09-11 | Biogen, Inc. | Methods of therapeutic administration of anti-cd40l compounds |
| WO1999000143A1 (en) * | 1997-06-27 | 1999-01-07 | Biogen, Inc. | Cd154 blockade therapy for autoimmune diseases |
-
1999
- 1999-04-02 CN CNB998067563A patent/CN1173735C/en not_active Expired - Fee Related
- 1999-04-02 JP JP2000542029A patent/JP2002510643A/en not_active Withdrawn
- 1999-04-02 AU AU33796/99A patent/AU743824B2/en not_active Withdrawn - After Issue
- 1999-04-02 KR KR1020007011045A patent/KR20010072564A/en active Search and Examination
- 1999-04-02 WO PCT/US1999/007321 patent/WO1999051258A1/en not_active Application Discontinuation
- 1999-04-02 EP EP99915237A patent/EP1067954A4/en not_active Withdrawn
- 1999-04-02 HU HU0101689A patent/HUP0101689A3/en unknown
- 1999-04-02 CN CNB200410078988XA patent/CN100352497C/en not_active Expired - Fee Related
- 1999-06-28 TW TW088105322A patent/TWI224969B/en not_active IP Right Cessation
-
2000
- 2000-10-02 NO NO20004966A patent/NO20004966L/en not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| NO20004966L (en) | 2000-12-04 |
| NO20004966D0 (en) | 2000-10-02 |
| KR20010072564A (en) | 2001-07-31 |
| EP1067954A1 (en) | 2001-01-17 |
| HUP0101689A3 (en) | 2004-08-30 |
| CN1757413A (en) | 2006-04-12 |
| CN100352497C (en) | 2007-12-05 |
| WO1999051258A1 (en) | 1999-10-14 |
| EP1067954A4 (en) | 2004-08-18 |
| TWI224969B (en) | 2004-12-11 |
| JP2002510643A (en) | 2002-04-09 |
| HUP0101689A2 (en) | 2001-08-28 |
| AU743824B2 (en) | 2002-02-07 |
| CN1303301A (en) | 2001-07-11 |
| AU3379699A (en) | 1999-10-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP4148537B2 (en) | CD40CR receptor and its ligand | |
| JP5802245B2 (en) | Human monoclonal antibody human CD134 (OX40) and methods for making and using the same | |
| JP2021036887A (en) | Humanized anti-ox40 antibodies and uses thereof | |
| Mathsson et al. | Immune complexes from rheumatoid arthritis synovial fluid induce FcγRIIa dependent and rheumatoid factor correlated production of tumour necrosis factor-α by peripheral blood mononuclear cells | |
| EP3882276A1 (en) | Bispecific antibody, preparation method therefor and application thereof | |
| RU2130782C1 (en) | Dna recombinant molecule encoding jcam-3 molecule, jcam-3 adhesion molecule, antibody able to bind with such molecule, pharmaceutical composition | |
| JP2018197236A (en) | Anti-tumour immune response to modified self-epitopes | |
| TWI222453B (en) | Ligand for herpes simplex virus entry mediator and methods of use | |
| CN106659775A (en) | Il-15-based molecules and methods of use thereof | |
| EA011302B1 (en) | Antibodies that bind interleukin-4 receptor | |
| EP0872488B1 (en) | MONOCLONAL ANTIBODY REACTING SPECIFICALLY WITH Fas LIGAND AND PROCESS FOR PRODUCING THE SAME | |
| EP1767625A1 (en) | Activity regulator for interferon producing cell | |
| AU2006202940A1 (en) | TR3-specific binding agents and methods for their use | |
| CN1173735C (en) | Use of anti-gp 39 antibodies for treatment and/or reversal of lupus and associated kidney disease | |
| AU2020387401A1 (en) | Anti-B7-H3 monoclonal antibody and methods of use thereof | |
| EP0740553B1 (en) | Monoclonal antibodies to antigens expressed by hematopoietic facilitatory cells | |
| ES2301164T3 (en) | SLAM, ANTIGEN COESTIMULATOR OF THE SURFACE OF CELLS T. | |
| US20020058037A1 (en) | Use of anti-gp-39 antibodies for treatment and/or reversal of lupus and lupus associated kidney disease |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20041103 |