Nothing Special   »   [go: up one dir, main page]

CN109254147A - Human peripheral blood immune cell function fully assesses kit and appraisal procedure - Google Patents

Human peripheral blood immune cell function fully assesses kit and appraisal procedure Download PDF

Info

Publication number
CN109254147A
CN109254147A CN201811191491.7A CN201811191491A CN109254147A CN 109254147 A CN109254147 A CN 109254147A CN 201811191491 A CN201811191491 A CN 201811191491A CN 109254147 A CN109254147 A CN 109254147A
Authority
CN
China
Prior art keywords
human
antibody
cell
ratio
fluorescein
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.)
Pending
Application number
CN201811191491.7A
Other languages
Chinese (zh)
Inventor
尹芝南
吴扬哲
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
DONGGUAN JINAN UNIVERSITY INSTITUTE
Original Assignee
DONGGUAN JINAN UNIVERSITY INSTITUTE
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by DONGGUAN JINAN UNIVERSITY INSTITUTE filed Critical DONGGUAN JINAN UNIVERSITY INSTITUTE
Priority to CN201811191491.7A priority Critical patent/CN109254147A/en
Publication of CN109254147A publication Critical patent/CN109254147A/en
Pending legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56966—Animal cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/7051—T-cell receptor (TcR)-CD3 complex
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70514—CD4
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70517—CD8
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70521—CD28, CD152
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70532—B7 molecules, e.g. CD80, CD86
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G01N2333/70539—MHC-molecules, e.g. HLA-molecules
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/7056—Selectin superfamily, e.g. LAM-1, GlyCAM, ELAM-1, PADGEM
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70578—NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30 CD40 or CD95
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70589—CD45
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70596—Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/715—Assays involving receptors, cell surface antigens or cell surface determinants for cytokines; for lymphokines; for interferons

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Hematology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Biomedical Technology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Virology (AREA)
  • Zoology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The present invention provides a kind of human peripheral blood immune cell function and fully assesses kit and appraisal procedure.It includes anti-human CD3, CD4, CD8, CD19, CD21, CD24, CD25, CD27, CD28, CD38, CD56, CD57, CD94, CD127, CD45RA, CXCR3, CXCR5, CCR4, CCR6, CCR7, HLA-DR, PD-1, P30, P46, NKG2D, KIR (NKB1), γ δ, V δ 2, IgD, IgM antibody that human peripheral blood immune cell function of the invention, which fully assesses kit, and the above antibody carries fluorescein label.The kit can be used in fully assessing the function of the immunocyte in human peripheral blood, easy to use, safe.

Description

Human peripheral blood immune cell function fully assesses kit and appraisal procedure
Technical field
The present invention relates to medical field more particularly to a kind of human peripheral blood immune cell function fully assess kit and Appraisal procedure.
Background technique
The generation of the immune function of human immunocyte and various diseases, progress have a directly related property, such as tumour Occur, development process is exactly because of the result that human autoimmune's cell function is lowered or is suppressed for a long time.But make a general survey of state Inside and outside Research Literature, discovery have not been used in the immunocyte that special assessment human peripheral blood has killing ability in the world at present The method of function.Clinical, third party testing agency evaluation immune function of human body method also rests on cell both at home and abroad at present The conventional indexs such as quantity, ratio.For example, the blood routine of hospital simply shows the quantitative range and ratio of leucocyte, as CD3 is thin Quantity, the ratio of born of the same parents, quantity, ratio of CD4+T cell etc., these data carry out the participation meaning of clinical diagnosis not to doctor Greatly.
Currently, including clinical and third party testing agency etc. without can really evaluate immunocyte function itself and quality Method.
Summary of the invention
The purpose of the present invention, which first consists in, provides a kind of human peripheral blood immune cell function comprehensive assessment kit, can It is easy to use, safe in conjunction with flow cytometer for being fully assessed to the function of the immunocyte in human peripheral blood.
The object of the invention is also to provide a kind of human peripheral blood immune cell functions to fully assess method, can be to people The function of immunocyte in peripheral body is fully assessed.
In order to achieve the above object, present invention firstly provides a kind of human peripheral blood immune cell functions to fully assess reagent Box, comprising: resist with fluorescein-labeled anti-CD3antibody, with fluorescein-labeled anti-human CD4 antibody, with fluorescein-labeled People CD8 antibody is marked with fluorescein-labeled anti human CD 19 antibody, with fluorescein-labeled anti-human CD21 antibody, band fluorescein Anti-human CD24 antibody, with fluorescein-labeled anti-human CD25 antibody, with fluorescein-labeled anti-human CD27 antibody, band fluorescein Label anti-human CD28 antibody, with fluorescein-labeled anti-human CD38 antibody, with fluorescein-labeled anti-human CD56 antibody, with glimmering The anti-human CD57 antibody of light element label resists with fluorescein-labeled anti-human CD94 antibody, with fluorescein-labeled anti-human CD127 Body resists with fluorescein-labeled anti-human CD 45 RA antibody, with fluorescein-labeled anti-human CXCR3 antibody, with fluorescein-labeled People CXCR5 antibody, with fluorescein-labeled anti-human CCR4 antibody, with fluorescein-labeled anti-human CCR6 antibody, band fluorescein mark Note anti-human CCR7 antibody, with fluorescein-labeled anti-human HLA-DR antibodies, with fluorescein-labeled anti-human PD-1 antibody, with glimmering The anti-human NKP30 antibody of light element label resists with fluorescein-labeled anti-human NKP46 antibody, with fluorescein-labeled anti-human NKG2D Body, with fluorescein-labeled anti-human KIR antibody, with fluorescein-labeled anti-human γ anti-δ, with fluorescein-labeled anti-human V δ 2 Antibody, with fluorescein-labeled anti-human IgD antibody, with fluorescein-labeled anti-human IgM antibodies.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are monoclonal antibody.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, any one in anti-human IgM antibodies are pulvis or liquid preparation.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are all made of the encapsulation of antibody reagent pipe, and the antibody reagent pipe is light transmittance In 10% brown plastic tube below.
Preferably, the light transmittance of the antibody reagent pipe is zero, i.e., completely opaque.
Optionally, when the antibody contained in the antibody reagent pipe is liquid preparation, antibody in the antibody reagent pipe Volume is 0.5ml-5ml.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are liquid preparation.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody At least one of 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are pulvis, the human peripheral blood immunocyte function It further includes phosphate buffer that kit, which can be fully assessed,.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies fluorescein label selected from PerCP-Cy5.5, BV510, FITC, AF647、PE-Cy7、BV421、PE、AF647、APC-H7、BB515。
The present invention also provides a kind of human peripheral blood immune cell functions to fully assess method, includes the following steps:
Step 1 provides human peripheral blood immune cell function comprehensive assessment kit as described above；
Peripheral blood mononuclear cells is extracted from 2ml-10ml human peripheral blood；
Step 2, when the human peripheral blood immune cell function fully assess kit in all antibody be liquid When preparation, takes 1 μ l-5 μ l respectively from all antibody, all mixed with the peripheral blood mononuclear cells, at 2 DEG C -6 DEG C Under the conditions of be protected from light incubation 10 minutes to 30 minutes, obtain cell detection sample；
It is first when at least a kind of antibody is pulvis in human peripheral blood immune cell function comprehensive assessment kit Pulvis is first dissolved as by antibody-solutions using phosphate buffer, takes 1 μ l-5 μ respectively from the antibody of all liq form later L is all mixed with the peripheral blood mononuclear cells, and incubation 10 minutes to 30 minutes is protected from light under the conditions of 2 DEG C -6 DEG C, is obtained To cell detection sample；
The concentration of antibody is 0.1-2mg/ml in the liquid preparation；The pulvis obtains after mixing with phosphate buffer Antibody-solutions in antibody concentration be 0.1-2mg/ml；
Step 3 tests and analyzes the cell detection sample using flow cytometer.
Beneficial effects of the present invention:
Human peripheral blood immune cell function of the invention fully assess kit can in conjunction with flow cytometer for pair The function of immunocyte in human peripheral blood is fully assessed, easy to use, and safe to the human body, which can be used for In the panimmunities functional assessment projects such as the immune function of human immunity health control evaluation and infections relating patient assessment.This The human peripheral blood immune cell function of invention fully assess method can function to the immunocyte in human peripheral blood into Row fully assesses, and operating procedure is simple, safe operation process.
Interior establish for the first time carries out comprehensively the function of the immunocyte in human peripheral blood to the present invention at the international level The method of assessment is fully assessed using function of the panimmunity functional parameter to the immunocyte in human peripheral blood, If judging, the function of the immunocyte in human peripheral blood is too low, prompts detected person using intervention means pair such as immunization therapies The immune function of itself is promoted, and the health status of human body is promoted.
Specific embodiment
Term as used herein:
" by ... preparation " it is synonymous with "comprising".Term "comprising" used herein, " comprising ", " having ", " containing " Or its any other deformation, it is intended that cover non-exclusionism includes.For example, composition, step, method comprising listed elements, Product or device are not necessarily limited to those elements, but may include not expressly listed other elements or such composition, step Suddenly, method, product or the intrinsic element of device.
Conjunction " by ... form " exclude any element that do not point out, step or component.If in claim, This phrase will make claim closed, so that it is not included the material in addition to the material of those descriptions, but relative Except customary impurities.When phrase " by ... form " be rather than immediately following theme in the clause that appears in claim main body after When, only it is limited to element described in the clause；Other elements be not excluded the claim as a whole it Outside.
Equivalent, concentration or other values or parameter are excellent with range, preferred scope or a series of upper limit preferred values and lower limit When the Range Representation that choosing value limits, this should be understood as specifically disclosing by any range limit or preferred value and any range Any pairing of lower limit or preferred value is formed by all ranges, regardless of whether the range separately discloses.For example, when open When range " 1~5 ", described range should be interpreted as including range " 1~4 ", " 1~3 ", " 1~2 ", " 1~2 and 4~ 5 ", " 1~3 and 5 " etc..When numberical range is described herein, unless otherwise stated, otherwise the range is intended to include its end Value and all integers and score in the range.
" mass parts " refer to the basic measurement unit for indicating the mass ratio relationship of multiple components, and 1 part can indicate arbitrary list Position quality, can such as be expressed as 1g, may also indicate that 2.689g etc..If we say that the mass parts of component A are a parts, the matter of B component Measuring part is b parts, then it represents that the quality of component A and the mass ratio a:b of B component.Alternatively, indicating that the quality of component A is aK, B group The quality divided is bK (K is arbitrary number, indicates multiplying factor).It can not misread, unlike mass fraction, all components The sum of mass parts be not limited to 100 parts of limitation.
"and/or" is used to indicate that one of illustrated situation or both may to occur, for example, A and/or B includes (A And B) and (A or B)；
In addition, indefinite article "an" before element of the present invention or component and "one" quantitative requirement to element or component (i.e. frequency of occurrence) unrestriction.Therefore "one" or "an" should be read as including one or at least one, and odd number The element or component of form also include plural form, unless the obvious purport of the quantity refers to singular.
Present invention firstly provides a kind of human peripheral blood immune cell functions to fully assess kit, comprising: band fluorescein The anti-CD3antibody of label, with fluorescein-labeled anti-human CD4 antibody, with fluorescein-labeled anti-human CD8 antibody, band fluorescence Element label anti human CD 19 antibody, with fluorescein-labeled anti-human CD21 antibody, with fluorescein-labeled anti-human CD24 antibody, band Fluorescein-labeled anti-human CD25 antibody resists with fluorescein-labeled anti-human CD27 antibody, with fluorescein-labeled anti-human CD28 Body, with fluorescein-labeled anti-human CD38 antibody, with fluorescein-labeled anti-human CD56 antibody, with fluorescein-labeled anti-human CD57 antibody is marked with fluorescein-labeled anti-human CD94 antibody, with fluorescein-labeled anti-human CD127 antibody, band fluorescein Anti-human CD 45 RA antibody, with fluorescein-labeled anti-human CXCR3 antibody, with fluorescein-labeled anti-human CXCR5 antibody, with glimmering Light element label anti-human CCR4 antibody, with fluorescein-labeled anti-human CCR6 antibody, with fluorescein-labeled anti-human CCR7 antibody, With fluorescein-labeled anti-human HLA-DR antibodies, with fluorescein-labeled anti-human PD-1 antibody, with fluorescein-labeled anti-human NKP30 antibody, with fluorescein-labeled anti-human NKP46 antibody, with fluorescein-labeled anti-human NKG2D antibody, band fluorescein mark Note anti-human KIR antibody, with fluorescein-labeled anti-human γ anti-δ, with fluorescein-labeled anti-human 2 antibody of V δ, band fluorescein Label anti-human IgD antibody, with fluorescein-labeled anti-human IgM antibodies.
Human peripheral blood immune cell function of the invention fully assess kit can in conjunction with flow cytometer for pair Immunocyte in human peripheral blood is tested and analyzed, and 72 immune functions of adaptive immune cell, described 72 are exempted from Epidemic disease functional parameter includes: T cell toatl proportion, the ratio of helper T lymphocyte, the ratio of killer T cell, double positive T cells The ratio (Th/Tc) of ratio, helper T lymphocyte and killer T cell, the ratio for initializing CD4+T cell, terminal differentiation CD4+ The ratio of T cell, the ratio of center memory CD4+T cell, the ratio of responsiveness memory CD4+T cell, failure CD4+T cell Ratio, the ratio of functionality CD4+T cell, the ratio of regulatory T cells, ratio, the terminal differentiation for initializing CD8+T cell The ratio of CD8+T cell, the ratio of center memory CD8+T cell, the ratio of responsiveness memory CD8+T cell, failure CD8+T are thin The ratio of born of the same parents, the ratio of inactive phase specific C D8+T cell, the ratio of inactive phase specificity terminal differentiation CD8+T cell, The ratio of the ratio of continuous expression virus-specific CD8+T cell, the terminal differentiation CD8+T cell of continuous expression virus-specific It is example, the ratio of the ratio of function interdiction CD8+T cell, the ratio of functionality CD8+T cell, terminal aging CD8+T cell, total Memorability CD8+T cell ratio, return the ratio of nest Memorability CD8+T cell, the ratio of Th1 cell, Th2 cell ratio Example, the ratio of Th17 cell, Th1 cell and the ratio (Th1/Th2) of Th2 cell, the ratio of Th17 cell and Th2 cell (Th17/Th2), the ratio (Th1+Th17/Th2) of Th1 cell+Th17 cell and Th2 cell, the ratio of Tfh cell, function suppression The ratio of property Tfh cell processed, the periphery auxiliary ratio of Tfh cell, the ratio of Tfh1 cell, the ratio of Tfh2 cell, Tfh17 are thin The ratio of born of the same parents, the ratio of Tc1 cell (1 type CD8+T cell), the ratio of Tc2 cell (2 type CD8+T cell), Tc17 cell (17 Type CD8+T cell) ratio, the ratio of NK T cell, the ratio of NK cell, the ratio of prematurity NK cell, maturation NK cell Ratio, prematurity/maturation NK ratio, the ratio of early function blocking property NK cell, advanced stage function of (Mature NK cells) Can the ratio of blocking property NK cell, the ratio of activated NK, ratio, the virus infection of conventional lethal NK cell specifically kill The ratio of wound property NK cell, the ratio of gamma delta T cells, the ratio of 1 positive T cell of V δ, the ratio of 2 positive T cell of V δ, V δ 1+/V δ The ratio of 2+, the ratio of 2 positive T cell of functionality V δ, the ratio of function inhibitio V δ 2+T cell, conventional 2 sun of lethal V δ The property ratio of T cell, the ratio of 2 positive T cell of virus infection specific killing V δ, the ratio of 1 positive T cell of functionality V δ, The ratio of 1 positive T cell of function inhibitio V δ, the ratio of conventional lethal 1 positive T cell of V δ, virus infection specific killing Property 1 positive T cell of V δ ratio, the ratio of B cell, non-conversion hysteria B cell (the memory unswitched (MZ-like) of memory B cells) ratio, the ratio of B cell (classic memory switched B cells) of memory conversion, thick liquid cell (Plasmablast) ratio of ratio, CD21 negative B cell (CD21low B cells), B progenitor cells (B Cells ratio), preceding juvenile cell (B cells) ratio, the ratio of B10 cell (B10cells), memory The ratio of property B cell (Memory B cells).
T cell toatl proportion is the ratio of T cell sum and total number of lymphocytes in the human peripheral blood of same volume；T is thin The effect of born of the same parents is to play adaptive immunity function；The term of reference of T cell toatl proportion is 45.76%~77.45%.
The ratio of helper T lymphocyte is the ratio of helper T lymphocyte number and T cell sum in the human peripheral blood of same volume Value；Helper T lymphocyte can play key effect in adaptive immunity, as antibody generates, the phagocytic function of macrophage, kills Signal need to be transmitted by helper T lymphocyte by hurting lethal effect of cell etc.；The term of reference of the ratio of helper T lymphocyte be 40%~ 67%.
The ratio of killer T cell is the ratio of killer T cell number and T cell sum in the human peripheral blood of same volume Value；Killer T cell mainly plays lethal immune function, and the cell such as cancer cell including removing mutation is removed by germ sense The cell of dye is purged to special antigen reactive target cell is generated；The term of reference of the ratio of killer T cell is 31%~58%.
The ratio of double positive T cells is the ratio of double positive T cell numbers and T cell sum in the human peripheral blood of same volume Value；Double positive T cells are mainly undifferentiated thymus T cells；The term of reference of the ratio of double positive T cells is 1%~5%.
The ratio (Th/Tc) of helper T lymphocyte and killer T cell is complementary T in the human peripheral blood of same volume The ratio of cell number and killer T cell number；The term of reference of the ratio (Th/Tc) of helper T lymphocyte and killer T cell is 0.57%~2.44%, it is clinical judgment immune function of human body disorder that the ratio is significant in immune function assessment Sensitive indicator shows that immune function is in overacfivity state and answers such as without acute infection or allergy if CD4/CD8 ratio increases The autoimmune diseases such as vigilant hypertension, diabetes and quasi-wind gateway then illustrate that body is in immunosupress shape if ratio reduces State, to be vigilant malignant tumour includes leukaemia, lupus erythematosus, AIDS, respiratory tract infection etc..
The ratio for initializing CD4+T cell is that CD4+T cell number and CD4+T are initialized in the human peripheral blood of same volume The ratio of total number of cells；Initialization CD4+T cell identifies the pathogen that immune system was never met and generates immune answer Answer reaction, the proportional representation of the cell initial credit of CD4+T cell and subsequent differentiation capability, after receiving antigenic stimulus, just Beginningization CD4+T is divided into different subtype cell at different conditions, plays different immune functions；Initialize the ratio of CD4+T cell The term of reference of example is 36.3% ± 15.3%.
The ratio of terminal differentiation CD4+T cell be same volume human peripheral blood in terminal differentiation CD4+T cell number with The ratio of CD4+T total number of cells；Terminal differentiation CD4+T cell is the CD4+T cell without differentiation updating ability；Terminal differentiation CD4+ The term of reference of the ratio of T cell is 15.3% ± 1.5%.
Center remember CD4+T cell ratio be same volume human peripheral blood in center remember CD4+T cell number with The ratio of CD4+T total number of cells；Center memory CD4+T cell is highly sensitive to antigenic stimulus (especially secondary stimulus), generates It is particularly important to generate immunological memory effect to vaccine for quick immunological effect reaction；Remember the ginseng of the ratio of CD4+T cell in center Examining range is 33.8% ± 15.6%.
The ratio that responsiveness remembers CD4+T cell is that responsiveness remembers CD4+T cell in the human peripheral blood of same volume Several ratios with CD4+T total number of cells；Responsiveness remember CD4+T cell have effector function, can all kinds of inflammation of quick release because Son；The term of reference that responsiveness remembers the ratio of CD4+T cell is 27.6%~55.9%.
The ratio of failure CD4+T cell is failure CD4+T cell number and CD4+T cell in the human peripheral blood of same volume The ratio of sum；Failure CD4+T cell number is associated with various autoimmune disease, such as type 1 diabetes, rheumatoid etc.；It declines The term of reference for exhausting the ratio of CD4+T cell is 11.96% ± 8.60%.
The ratio of functional CD4+T cell is functionality CD4+T cell number and CD4+T in the human peripheral blood of same volume The ratio of total number of cells；Functional CD4+T cell is can reactivity CD4+T cell；The reference of the ratio of functional CD4+T cell Range is 89.56% ± 3.06%.
The ratio of regulatory T cells is regulatory T cells number and CD4+T total number of cells in the human peripheral blood of same volume Ratio；Regulatory T cells maintain immunologic balance important adjusting cell, mainly to internal autoimmune response rise negative control/ Inhibiting effect regulates and controls immune tolerance.Its ratio plays the role of tumour, immune deficiency, repulsion very big.It is excessively high with tumour phase It closes, it is too low related to autoimmunity disease；The term of reference of the ratio of regulatory T cells is 4.14% ± 1.85%.
The ratio for initializing CD8+T cell is that CD8+T cell number and CD8+T are initialized in the human peripheral blood of same volume The ratio of total number of cells；Initializing CD8+T cell is one of important cells group of cellular immunotherapy；Initialize CD8+T cell Ratio term of reference be 35.4%~53.2%.
The ratio of terminal differentiation CD8+T cell be same volume human peripheral blood in terminal differentiation CD8+T cell number with The ratio of CD8+T total number of cells；Terminal differentiation CD8+T cell is the cd8 cell without differentiation more new function；Terminal differentiation CD8+T The term of reference of the ratio of cell is 21.2%~40.3%.
Center remember CD8+T cell ratio be same volume human peripheral blood in center remember CD8+T cell number with The ratio of CD8+T total number of cells；Center memory CD8+T cell is to virus, antigen subinfection or the sensitive cell of stimulation again, energy Tachysynthesis response is enough transferred, proliferation can be stablized in secondary lymphatic organ and is divided into effector cell under antigenic stimulus；Center The term of reference for remembering the ratio of CD8+T cell is 4.61%~6.0%.
The ratio that responsiveness remembers CD8+T cell is that responsiveness remembers CD8+T cell in the human peripheral blood of same volume Several ratios with CD8+T total number of cells；Responsiveness, which remembers CD8+T cell, has anti-infective, anti-tumor function, releasable a variety of anti- Virocyte cerebroysin simultaneously has killing ability to mutation cell；The term of reference of ratio of responsiveness memory CD8+T cell is 33.1%~56.3%.
The ratio of failure CD8+T cell is failure CD8+T cell number and CD8+T cell in the human peripheral blood of same volume The ratio of sum；Failure CD8+T cell number is associated with a variety of viral infections.If failure cell increases, show cd8 cell Anti-infective, antitumor and removing target cell ability reduces；The term of reference of the ratio of failure CD8+T cell is 30%~55%.
The ratio of inactive phase specific C D8+T cell is inactive phase specificity in the human peripheral blood of same volume The ratio of CD8+T cell number and responsiveness memory CD8+T total number of cells；The ginseng of the ratio of inactive phase specific C D8+T cell Examining range is 78.7%~96%.
The ratio of inactive phase specificity terminal differentiation CD8+T cell is the inactive phase in the human peripheral blood of same volume The ratio of specific terminal differentiation CD8+T cell number and terminal differentiation CD8+T total number of cells；Inactive phase specificity terminal differentiation The term of reference of the ratio of CD8+T cell is 85.2%~98.8%.
The ratio of continuous expression virus-specific CD8+T cell is continuous expression virus in the human peripheral blood of same volume The ratio of specific C D8+T cell number and responsiveness memory CD8+T total number of cells；Continuous expression virus-specific CD8+T cell Ratio term of reference be 2.3%~8.6%.
The ratio of the terminal differentiation CD8+T cell of continuous expression virus-specific be same volume human peripheral blood in hold The terminal differentiation CD8+T cell number of continued da virus specificity and the ratio of terminal differentiation CD8+T total number of cells；It is described to continue table The term of reference of the ratio of the terminal differentiation CD8+T cell of da virus specificity is 6.4%~20%.
The ratio of function interdiction CD8+T cell is function interdiction CD8+T cell in the human peripheral blood of same volume Several ratios with CD8+T total number of cells.Function interdiction CD8+T is the CD8+T cell of function inhibitio, especially with it is antitumor Ability is related, is maintaining body's immunity stable state and is establishing in immune tolerance to play an important role, can be used as The immune detection index of autoimmune disease, AIDS and immune deficiency, ratio increase, then immunological effect effect is pressed down System.The term of reference of the ratio of function interdiction CD8+T cell is 24.4 ± 7.6%.
The ratio of functional CD8+T cell is functionality CD8+T cell number and CD8+T in the human peripheral blood of same volume The ratio of total number of cells.Functional CD8+T cell is to kill sick germ infection with the active CD8+T cell of immune function Target cell and tumour cell, important effector function is played in antitumor, anti-infective and allograft rejection.Function Property CD8+T cell ratio term of reference be 52.5%-70%.
The ratio of terminal aging CD8+T cell be same volume human peripheral blood in terminal aging CD8+T cell number with The ratio of CD8+T total number of cells.Terminal aging CD8+T cell has the characteristics that low proliferation, easy apoptosis, and ratio raising is exempted from it The decline of epidemic disease effect is related.The term of reference of the ratio of terminal aging CD8+T cell is 5.45%~20.48%.
The ratio of total Memorability CD8+T cell is Memorability CD8+T cell total in the human peripheral blood of same volume Several ratios with CD8+T total number of cells.Total Memorability CD8+T cell is can be stimulated again by antigen and quickly activate and play straight Connect the subgroup of killing tumor cell or germ infection cell.The term of reference of the ratio of total Memorability CD8+T cell is 12.9%~25.8%.
The ratio for returning nest Memorability CD8+T cell is that nest Memorability CD8+T is returned in the human peripheral blood of same volume The ratio of cell number and total Memorability CD8+T cell number.Returning nest Memorability CD8+T cell is that can move to second level leaching Bar organ receives antigenic stimulus and re-activation is effector cell, is the important indicator of the treatment and prognosis of HIV infection patient.It returns The term of reference of the ratio of nest Memorability CD8+T cell is 73% ± 10.3%.
The ratio of Th1 cell is the ratio of Th1 cell number and Th total number of cells in the human peripheral blood of same volume.Th1 Cell is also known as inflammatory Tcell, Major Secretory interleukin-22, interferon-γ, interferon-a, tumor necrosis factor etc., main to be situated between The immune response of guided cell poison and local inflammation-related, auxiliary antibody generate, and participate in cellular immunity and delayed type hypersensitivity inflammation Generation, delayed hypersensitivity T cell can be considered as.The participation of Th1 cell resists germ intracellular immune.The ratio mistake of Th1 cell Height can cause organ specific autoimmune's disease.The term of reference of the ratio of Th1 cell is 14%~46%.
The ratio of Th2 cell is the ratio of Th2 cell number and Th total number of cells in the human peripheral blood of same volume.Th2 Cell Major Secretory IL-4,5,6 and 10 etc., major function is stimulation B cell proliferation and generates immunoglobulin G (IgG1) It is related with humoral immunity with E (IgE) antibody.The participation of Th2 cell resists extracellular cause of disease parasite immunity.The ratio of Th2 cell is excessively high It can cause allergic inflammation and asthma.The term of reference of the ratio of Th2 cell is 12%~25%.
The ratio of Th17 cell is the ratio of Th17 cell number and Th total number of cells in the human peripheral blood of same volume. Th17 cell participation mediating inflammatory reaction (resisting extracellular bacterium, fungal infection), autoimmune disease, tumour and graft rejection etc. Occurrence and development.Th17 cell mainly plays pro-inflammatory effect, and ratio is excessively high to cause diseases associated with inflammation (EAE, CIA etc.).Th17 The term of reference of the ratio of cell is 0.8%~8.3%.
The ratio (Th1/Th2) of Th1 cell and Th2 cell is Th1 cell number and Th2 in the human peripheral blood of same volume The ratio of cell number；The term of reference of the ratio (Th1/Th2) of Th1 cell and Th2 cell is 0.71~3.33；Th1/Th2 ratio Example, which increases, will promote cell immune response, enhance the activity of CD+8T cell, enhance disease in anti-tumor capacity and scavenger-cell The ability of poison, but increase inflammatory reaction simultaneously, as hepatitis B virus infection can aggravate the damage of liver cell.Th1/Th2 ratio reduces It will promote humoral immune reaction, but inhibit cell immune response, cytotoxic T lymphocyte (cytotoxicity Tlymphocyte, CTL) cell activity decrease, hepatocellular injury mitigation.
The ratio (Th17/Th2) of Th17 cell and Th2 cell be in the human peripheral blood of same volume Th17 cell number with The ratio of Th2 cell number；The term of reference of the ratio (Th17/Th2) of Th17 cell and Th2 cell is 0.04~0.6；Th17/ The raising of Th2 ratio occurs to diseases associated with inflammation, the tumour etc. including chronic inflammation, development is related.
The ratio (Th1+Th17/Th2) of Th1 cell+Th17 cell and Th2 cell is in the human peripheral blood of same volume The ratio of Th1 cell+Th17 cell number and Th2 cell number；The ratio (Th17/Th2) of Th1 cell+Th17 cell and Th2 cell Term of reference be 0.89~3.79；The diseases such as Th1+Th17/Th2 and autoimmune disease, tumour and graft-versus-host Occurrence and development it is related.
The ratio of Tfh cell is the ratio of Tfh cell number and CD4+T total number of cells in the human peripheral blood of same volume. Tfh cell secretes cytokine profiles and a variety of costimulatory molecules of expression complete the booster action that antibody is generated to B cell.Tfh The ratio of cell is excessively high to cause antibody associated autoimmune disease.The ratio of Tfh cell is too low to cause immune deficiency.Th17 The term of reference of the ratio of cell is 23%~41.2%.
The ratio of function inhibitio Tfh cell be same volume human peripheral blood in function inhibitio Tfh cell number with The ratio of CD4+T total number of cells.Function inhibitio Tfh cell has immune down regulation, and it is living to can inhibit Tfh cell transition Change.The term of reference of the ratio of function inhibitio Tfh cell is 15.2%~36%.
The ratio of periphery auxiliary Tfh cell is periphery auxiliary Tfh cell number and CD4+T in the human peripheral blood of same volume The ratio of total number of cells.Periphery auxiliary Tfh cell is with the T helper cell for inhibiting function.The ratio of periphery auxiliary Tfh cell Term of reference be 22.6%~39.6%.
The ratio of Tfh1 cell is the ratio of Tfh1 cell number and Tfh total number of cells in the human peripheral blood of same volume. Tfh1 cell does not play a major role in Help B Cells differentiation, poor related to the antibody response of influenza vaccines.Tfh1 cell exists Quantity is reduced in the autoimmunity diseases such as spontaneous lymthoma.The term of reference of the ratio of Tfh1 cell is 19.2%~29.8%.
The ratio of Tfh2 cell is the ratio of Tfh2 cell number and Tfh total number of cells in the human peripheral blood of same volume. Tfh2 cell more effectively can assist naive B cell to be differentiated to form plasmablast, non-suddenly in autoimmunity disease and high-level B cell Quantity increases in the patients such as golden surprise lymthoma.The term of reference of the ratio of Tfh2 cell is 15.3%~23.6%.
The ratio of Tfh17 cell is the ratio of Tfh17 cell number and Tfh total number of cells in the human peripheral blood of same volume Value.Tfh17 cell effectively can assist naive B cell to be differentiated to form plasmablast as Tfh2 cell.The function of Tfh17 cell It can be similar with the function of Tfh2 cell.The term of reference of the ratio of Tfh17 cell is 31%~40%.
The ratio of Tc1 cell (1 type CD8+T cell) is Tc1 cell number and Tc cell in the human peripheral blood of same volume The ratio of sum.Tc1 cell is 1 type cytotoxic T cell, can migrate to inflammation part, secrete INF- γ, kill target cell, reach To the purpose of viral infection resisting.
The ratio of Tc2 cell (2 type CD8+T cell) is Tc2 cell number and Tc cell in the human peripheral blood of same volume The ratio of sum.Tc2 cell is 2 type cytotoxic T cells, secretes IL4, IL5, IL10.Tc2 cell mainly with chronic infection, swollen The chronic diseases such as tumor, autoimmune disease are related.This group of cytosis are related to the severity of disease and progress.
The ratio of Tc17 cell (17 type CD8+T cell) is that Tc17 cell number and Tc are thin in the human peripheral blood of same volume The ratio of born of the same parents' sum.Tc17 cell is a group cytotoxic T cell for generating interleukin-17, is sent out in antitumor, anti-infectious immunity Wave important function.Tc17 cell plays a significant role in anti-influenza A virus together with Tc1 cell.
The ratio of NK T cell is the ratio of NK T cell number and total number of lymphocytes in the human peripheral blood of same volume. NK T cell is with the active one kind NK cell of direct killing.The term of reference of the ratio of NK T cell be 11.71%~ 33.1%.
The ratio of NK cell is the ratio of NK cell number and total number of lymphocytes in the human peripheral blood of same volume.NK is thin Born of the same parents are the important immunocytes of human body, with antitumor, viral infection resisting and immunological regulation have it is close contact, also participate in super quick The generation of reaction and autoimmune disease can identify target cell, killing medium.The term of reference of the ratio of NK cell is 6.26%~37.16%.
The ratio of prematurity NK cell is prematurity NK cell number and NK total number of cells in the human peripheral blood of same volume Ratio.Prematurity NK cell is a subgroup of NK cell, and mature NK cell ratio, to the killing ability of target cell compared with It is low, but most can efficiently generate cell factor such as IFN-gamma.The term of reference of the ratio of prematurity NK cell be 5.6~ 27.8%.
The ratio of mature NK cell is the ratio of maturation NK cell number and NK total number of cells in the human peripheral blood of same volume Value.Mature NK cell has powerful killing activity from immature NK cell differentiation, can direct killing rake cell.Mature NK The term of reference of the ratio of cell is 43~82%.
Prematurity/maturation NK ratio is the ratio of prematurity NK cell number and maturation NK cell number.The ratio is with the state of an illness Development and curative effect have close ties, and ratio increases generally related to unsatisfactory curative effect or disease progression.The term of reference of the ratio is 0.068~0.72.
The ratio of early function blocking property NK cell is that early function blocking property NK is thin in the human peripheral blood of same volume The ratio of born of the same parents' number and NK total number of cells.Early function blocking property NK cell can reinforce the killing ability of NK cell.Early function The term of reference of the ratio of blocking property NK cell is 36.3%~52.7%.
The ratio of advanced stage function interdiction NK cell is that the human peripheral blood middle and advanced stage function interdiction NK of same volume is thin The ratio of born of the same parents' number and NK total number of cells.Advanced stage function interdiction NK cell is modulability NK cell.Advanced stage, function interdiction NK was thin The term of reference of the ratio of born of the same parents is 22%~29%.
The ratio of activated NK is the ratio of activated NK number and NK total number of cells in the human peripheral blood of same volume Value.Activated NK is that have antitumor, antiviral functions functional NK cells.The term of reference of the ratio of activated NK It is 25%~53%.
The ratio of conventional lethal NK cell is lethal NK cell number conventional in the human peripheral blood of same volume With the ratio of NK total number of cells.Conventional lethal NK cell is the effector cell of antineoplastic immune, mainly to tumour cell and The cell of germ infection plays killing or scavenging effect.The term of reference of the ratio of conventional lethal NK cell is 38.5%~ 64%.
The ratio of virus infection specific killing NK cell is that virus infection is specifically killed in the human peripheral blood of same volume The ratio of wound property NK cell number and NK total number of cells.Virus infection specific killing NK cell is directly related with virus infection, disease The ratio of poison infection specific killing NK cell reduces related to virus infection, tumour progression etc..Virus infection specific killing The term of reference of the ratio of NK cell is 9.93%~80.85%.
The ratio of gamma delta T cells is the ratio of gamma delta T cells number and T cell sum in the human peripheral blood of same volume.γδ T cell is the first line of defence for participating in immunity of organism, has certain non-specific lethal effect, and anti-swollen with wide spectrum Tumor, anti-infectious function.The term of reference of the ratio of gamma delta T cells is 0.5%~8.2%.
The ratio of 1 positive T cell of V δ is that 1 positive T cell number of V δ and gamma delta T cells are total in the human peripheral blood of same volume Several ratio.1 positive T cell of V δ is the γ delta cell subgroup with immunosupress, regulatory function.The ratio of 1 positive T cell of V δ Term of reference be 8.2%~24%.
The ratio of 2 positive T cell of V δ is that 2 positive T cell number of V δ and gamma delta T cells are total in the human peripheral blood of same volume Several ratio.2 positive T cell of V δ is mainly to play antitumor, anti-infectious function gamma delta T cells subgroup.2 positive T cell of V δ The term of reference of ratio is 46.8%~72%.
The ratio of V δ 1+/V δ 2+ is the ratio of 1 positive T cell number of V δ and 2 positive T cell number of V δ.The ratio and gamma delta T are thin The functional status of born of the same parents' subgroup is directly related, and ratio increases the function reduction for generally showing gamma delta T cells subgroup.1 positive T cell of V δ Several term of reference with the ratio of 2 positive T cell of V δ are 0.12~0.51.
The ratio of functional 2 positive T cell of V δ is 2 positive T cell number of functionality V δ in the human peripheral blood of same volume With the ratio of gamma delta T cells sum.The ratio with antitumor, anti-infective function is directly related.The ratio of functional 2 positive T cell of V δ The term of reference of example is 94.6%~99.8%.
The ratio of 2 positive T cell of function inhibitio V δ is that function inhibitio V δ 2 is positive in the human peripheral blood of same volume The ratio of T cell number and 2 gamma delta T cells of V δ sum.2 positive T cell of function inhibitio V δ can control cell transition activation, swell Ratio raising shows that anti-tumor function is suppressed in tumor patient.The term of reference of the ratio of function inhibitio V δ 2+T cell is 6.4%~38.9%.
The ratio of conventional lethal 2 positive T cell of V δ is lethal V δ 2 conventional in the human peripheral blood of same volume The ratio of positive T cell number and 2 gamma delta T cells of V δ sum.Conventional lethal 2 positive T cell of V δ can play antitumor, anti- Infection effect.The term of reference of the ratio of conventional lethal 2 positive T cell of V δ is 0~1%.
The ratio of 2 positive T cell of virus infection specific killing V δ is virus infection in the human peripheral blood of same volume The ratio of 2 positive T cell number of specific killing V δ and 2 gamma delta T cells of V δ sum.2 positive T of virus infection specific killing V δ is thin Born of the same parents are directly related with virus infection, and the ratio of 2 positive T cell of virus infection specific killing V δ reduces and virus infection, tumour The correlations such as progress, and influence the activation of cell.The term of reference of the ratio of 2 positive T cell of virus infection specific killing V δ is 0 ~0.8%.
The ratio of functional 1 positive T cell of V δ is in the human peripheral blood of same volume be same volume human peripheral The ratio of functionality V δ 1 positive T cell and 1 gamma delta T cells of V δ sum in blood.Functional 1 positive T cell of V δ is with antitumor The Vdelta1 cell mass of function.The term of reference of the ratio of functional 1 positive T cell of V δ is 94.6%~99.8%.
The ratio of 1 positive T cell of function inhibitio V δ is that function inhibitio V δ 1 is positive in the human peripheral blood of same volume The ratio of T cell number and 1 gamma delta T cells of V δ sum.1 positive T cell of function inhibitio V δ has function inhibitio Vdelta1 cell mass inhibits cell hyperactivity.The term of reference of the ratio of 1 positive T cell of function inhibitio V δ is 8.4% ~38.9%.
The ratio of conventional lethal 1 positive T cell of V δ is lethal V δ 1 conventional in the human peripheral blood of same volume The ratio of positive T cell number and 1 gamma delta T cells of V δ sum.Conventional lethal 1 positive T cell of V δ be can play it is antitumor, anti- The Vdelta1 cell mass of infection effect.The term of reference of the ratio of conventional lethal 1 positive T cell of V δ is 0~1%.
The ratio of 1 positive T cell of virus infection specific killing V δ is human peripheral in the human peripheral blood of same volume The ratio of 1 positive T cell number of virus infection specific killing V δ and 1 gamma delta T cells of V δ sum in blood.Virus infection specific killing Property V 1 positive T cell of δ it is directly related with virus infection, ratio reduce it is related to virus infection, tumour progression etc..Virus infection is special The term of reference of the ratio of different lethal 1 positive T cell of V δ is 0~9.6%.
The ratio of B cell is the ratio of B cell number and total number of lymphocytes in the human peripheral blood of same volume.B cell It is main to generate antibody, critical function is played in humoral immunity.The term of reference of the ratio of B cell is 5~15%.
The ratio for remembering non-conversion hysteria B cell (memory unswitched (MZ-like) B cells) is same volume Human peripheral blood in remember the ratio of non-conversion hysteria B cell number Yu B cell sum.Remembering non-conversion hysteria B cell is memory-type B Cell precursors, quantity representative can be converted to the ability of memory-type B cell.Remember the reference of the ratio of non-conversion hysteria B cell Range is 3.8%~35.6%.
The ratio for the B cell (classic memory switched B cells) converted is remembered into the people of same volume The B cell number of conversion and the ratio of B cell sum are remembered in peripheral body.The B cell of memory conversion belongs to switched memory Type B cell, surface Ig are transformed into from IgD for IgG, IgA or IgE, can be further to Plasmablast (plasmablast) Conversion, starts a large amount of secretory antibodies.The term of reference of the ratio for the B cell converted is remembered into 4.6%~35.5%.
The ratio of thick liquid cell (Plasmablast) is the human peripheral blood mesoplasmatocyte number and B cell sum of same volume Ratio.Thick liquid cell by mature B cell through antigenic stimulus activate come, major function is secretory antibody, neutralization antigen.Thick liquid cell Ratio term of reference be 0.3%~7.8%.
The ratio of CD21 negative B cell (CD21low B cells) is CD21 feminine gender B in the human peripheral blood of same volume The ratio of cell number and B cell sum.CD21 negative B cell is a kind of B cell of memory-type, in chronic inflammations such as autoimmunity diseases Quantity increases in disease patient.The term of reference of the ratio of CD21 negative B cell is 1.9%~19.3%.
B progenitor cells (B cells) ratio be same volume human peripheral blood in B progenitor cells number and B The ratio of total number of cells.B progenitor cells are not yet exposed or the B cell of contact antigen, can be broken up by antigenic stimulus and be generated not Same type B cell represents the reserve capabillity of B cell.The term of reference of the ratio of B progenitor cells is 17.9%~75.1%.
Preceding juvenile cell (B cells) ratio be same volume human peripheral blood in preceding juvenile cell Several ratios with B cell sum.Its ability for being converted into juvenile cell of the proportional representation of preceding juvenile cell.
The ratio of B10 cell (B10cells) is B10 cell number and B cell sum in the human peripheral blood of same volume Ratio.B10 cell is that a group plays immunological regulation and inhibits the B cell of function, IL-10 can be secreted, in tumor development process In to immune response rise inhibiting effect, can prevent from being immunoreacted the damage to body itself in autoimmune disease.
The ratio of memory B cells (Memory B cells) is memory B cells in the human peripheral blood of same volume Several ratios with B cell sum.Memory B cells are the important effect cells for generating immunological memory function, can be to antigen two Secondary stimulation quick response generates lot of antibodies.The term of reference of the ratio of memory B cells is 11%~46.6%.
Specifically, the term of reference mentioned in the application refers to the normal range (NR) of Healthy People.
Above-mentioned 72 immune functions can reflect the immune functional state of human peripheral blood T cell comprehensively, by right Above 72 immune functions are analyzed, if it is determined that human body immunologic hypofunction (such as initialization cell tail off, function Property cell tails off, failure cell increases), then prompt may need to carry out immunologic intervention, such as pass through drug, functional food or life The method of object treatment improves the immune function of body.
To obtain above-mentioned 72 immune functions, it is necessary first to extract peripheral blood mononuclear cells from human peripheral blood (PBMC), taken respectively from all antibody of this kit later a certain amount of, all mixed with peripheral blood mononuclear cells, It is incubated for, obtains cell detection sample, cell detection sample is detected using flow cytometer later, utilizes flow cytometer After included analysis software or third-party analysis software are for statistical analysis to the analysis result of various immunocytes, in acquisition State 17 immune functions.
Specifically, the cell sign of each immunocyte group are as follows:
The total group of T cell: CD3+；
Helper T lymphocyte: CD3+CD4+；
Killer T cell: CD3+CD8+；
Double positive T cells: CD3+CD4+CD8+；
Initialize CD4+T cell: CD3+CD4+CD45RA+CCR7+；
Terminal differentiation CD4+T cell: CD3+CD4+CD45RA+CCR7-；
Remember CD4+T cell: CD3+CD4+CD45RA-CCR7+ in center；
Responsiveness remembers CD4+T cell: CD3+CD4+CD45RA-CCR7-；
Failure CD4+T cell: CD3+CD4+CD28-；
Functional CD4+T cell: CD3+CD4+CD28+；
Regulatory T cells: CD3+CD4+CD25+CD127-；
Initialize CD8+T cell: CD3+CD8+CCR7+CD45RA+；
Terminal differentiation CD8+T cell: CD3+CD8+CCR7-CD45RA+；
Remember CD8+T cell: CD3+CD8+CCR7+CD45RA- in center；
Responsiveness remembers CD8+T cell: CD3+CD8+CCR7-CD45RA-；
Failure CD8+T cell: CD3+CD8+CD28-；
Inactive phase specific C D8+T cell: CD3+CD8+CCR7-CD45RA-CD127high；
Inactive phase specificity terminal differentiation CD8+T cell: CD3+CD8+CCR7-CD45RA+CD127high；
Continuous expression virus-specific CD8+T cell: CD3+CD8+CCR7-CD45RA-CD127low；
The terminal differentiation CD8+T cell of continuous expression virus-specific: CD3+CD8+CCR7-CD45RA+CD127low；
Function interdiction CD8+T cell: CD3+CD8+PD-1+；
Functional CD8+T cell: CD3+CD8+CD28+；
Terminal aging CD8+T cell: CD3+CD8+CD28-CD57+；
Total Memorability CD8+T cell: CD3+CD8+HLADR+；
Return nest Memorability CD8+T cell: CD3+CD8+HLADR+CD38+；
Th1 cell: CD3+CD4+CXCR5-CXCR3+CCR4-；
Th2 cell: CD3+CD4+CXCR5-CXCR3-CCR4+；
Th17 cell: CD3+CD4+CXCR5-CXCR3-CCR4-CCR6+；
Tfh cell: CD3+CD4+CXCR5+；
Function inhibitio Tfh cell: CD3+CD4+CXCR5+PD-1+；
Periphery auxiliary Tfh cell: CD3+CD4+CXCR5-PD-1+；
Tfh1 cell: CD3+CD4+CXCR5+CXCR3+CCR4-；
Tfh2 cell: CD3+CD4+CXCR5+CXCR3-CCR4+；
Tfh17 cell: CD3+CD4+CXCR5+CXCR3-CCR4-CCR6+；
Tc1 cell: CD3+CD8+CXCR5-CXCR3+CCR4-；
Tc2 cell: CD3+CD8+CXCR5-CXCR3-CCR4+；
Tc17 cell: CD3+CD8+CXCR5-CXCR3-CCR4-CCR6+；
NKT cell: CD3+CD56+；
NK cell: CD3-CD56+；
Prematurity NK cell: CD3-CD56+bright；
Mature NK cell: CD3-CD56+dim；
Early function blocking property NK cell: CD3-CD56+CD94+KIR-；
Advanced stage function interdiction NK cell: CD3-CD56+CD94-KIR+；
Activated NK: CD3-CD56+NKG2D+；
Conventional lethal NK cell: CD3-CD56+NKP30+；
Virus infection specific killing NK cell: CD3-CD56+NKP46+；
Gamma delta T cells: CD3+ γ δ+；
2 positive T cell of V δ: CD3+ γ δ+V δ 2+；
Functional 2 positive T cell of V δ: CD3+ γ δ+V δ 2+NKG2D+；
2 positive T cell of function inhibitio V δ: CD3+ γ δ+V δ 2+PD1+；
Conventional lethal 2 positive T cell of V δ: CD3+ γ δ+V δ 2+NKP30+；
2 positive T cell of virus infection specific killing V δ: CD3+ γ δ+V δ 2+NKP46+；
Functional 1 positive T cell of V δ: CD3+ γ δ+V δ 1+NKG2D+；
1 positive T cell of function inhibitio V δ: CD3+ γ δ+V δ 1+PD1+；
Conventional lethal 1 positive T cell of V δ: CD3+ γ δ+V δ 1+NKP30+；
1 positive T cell of virus infection specific killing V δ: CD3+ γ δ+V δ 1+NKP46+；
The total group of B cell: CD3-CD19+；
Remember non-conversion hysteria B cell: CD3-CD19+CD27+IgD+；
Remember the B cell of conversion: CD3-CD19+IgD-IgM-CD27+CD38low；
Thick liquid cell: CD3-CD19+IgD-IgM-CD27+CD38+；
CD21 negative B cell: CD3-CD19+IgD+CD27-CD38low CD21low；
B progenitor cells: CD3-CD19+CD27-IgD+；
Preceding juvenile cell: CD3-CD19+IgM-IgD-CD27-CD38+；
B10 cell: CD3-CD19+IgD+IgM+CD27+CD24+CD38-；
Memory B cells: CD3-CD19+IgD+IgM+CD27+CD38+CD24+.
Preferably, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are monoclonal antibody.This is because the specificity of monoclonal antibody is good, Fluorescence intensity after dyeing and antigenic expression are linear relationships, and the difference between monoclonal antibody difference production batch compared with It is small.Although polyclonal antibody can produce stronger signal, but poor specificity, and fluorescence intensity is not with antigen levels after dyeing Linear relationship, and it is widely different between polyclonal antibody difference production batch, therefore Dan Ke is used during flow cytometer detection Grand antibody can guarantee higher detection accuracy.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, any one in anti-human IgM antibodies are pulvis or liquid preparation.
The pulvis needs to be added appropriate phosphate buffer (PBS) before use and is dissolved, and the liquid preparation is not It needs to dissolve, directly use.The pulvis can be placed under the conditions of 4 DEG C with liquid preparation and save, and pulvis can also be placed in -20 DEG C Under the conditions of save.
Specifically, in the liquid preparation antibody concentration be 0.1-2mg/ml (such as 0.1mg/ml, 0.5mg/ml, 1mg/ml、1.5mg/ml、2mg/ml)。
Optionally, the ingredient of the liquid preparation includes antibody and phosphate buffer.
Specifically, the pulvis is in use, be mixed to get antibody-solutions for the pulvis and phosphate buffer, it is described anti- The concentration of antibody is 0.1-2mg/ml (such as 0.1mg/ml, 0.5mg/ml, 1mg/ml, 1.5mg/ml, 2mg/ml) in liquid solution.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are all made of the encapsulation of antibody reagent pipe.
Preferably, the antibody reagent pipe is completely opaque brown plastic tube, in other words, the brown plastic tube Light transmittance be zero.
Optionally, when the antibody contained in the antibody reagent pipe is liquid preparation, antibody in the antibody reagent pipe Volume is 0.5ml-5ml, such as 0.5ml, 1ml, 2ml, 3ml, 4ml, 5ml etc., the use time of the antibody in the antibody reagent pipe Number is 50-500 times.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies are liquid preparation, and liquid preparation uses more convenient, can be saved Dissolving step before going pulvis use, and it is avoided that caused detection error problem when pulvis dissolution is uneven.
, it is understood that although the antibody of powder form uses comparatively laborious, the step of increasing dissolution is needed, but It is for the antibody compared to liquid forms, the shelf-life of the antibody of powder form is longer.
When the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, Anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 are anti- It is body, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, Anti-human NKP30 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human V δ 2 are anti- When at least one of body, anti-human IgD antibody, anti-human IgM antibodies are pulvis, the human peripheral blood immune cell function is comprehensive Assessing kit further includes phosphate buffer (PBS), and the phosphate buffer can be placed in reagent bottle.By providing phosphoric acid Salt buffer has saved the time for preparing phosphate buffer for user, keeps the use of antibody more convenient.
Optionally, the anti-CD3antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 It is antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD27 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, anti-human CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 are anti- It is body, anti-human CXCR5 antibody, anti-human CCR4 antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, resists anti-human NKP30 antibody 2 antibody of people V δ, anti-human IgD antibody, anti-human IgM antibodies fluorescein label selected from PerCP-Cy5.5, BV510, FITC, AF647、PE-Cy7、BV421、PE、AF647、APC-H7、BB515。
The present invention also provides a kind of human peripheral blood immune cell functions to fully assess method, includes the following steps:
Step 1 provides human peripheral blood immune cell function comprehensive assessment kit described above；
Peripheral blood mononuclear cells is extracted from 2ml-10ml (such as 2ml, 4ml, 6ml, 8ml, 10ml) human peripheral blood (PBMC)。
Since the particular content that the human peripheral blood immune cell function fully assesses kit has carried out in detail above Thin description, therefore details are not described herein again.
Optionally, the method that peripheral blood mononuclear cells (PBMC) is extracted from human peripheral blood includes the following steps:
1) whole blood sample of 2ml-10ml human peripheral blood is added in centrifuge tube；
2) isometric phosphate buffer (PBS) is added in whole blood sample, PBS+ whole blood sample is fluctuated mixed Uniformly, sample diluent is obtained；
3) new 15ml centrifuge tube is taken, FICOLL liquid and sample diluent is added, it is usually dilute according to FICOLL liquid and sample Release the ratio addition of the volume ratio 1:1 of liquid；
4) centrifuge tube is tilted 45 °, sample diluent is carefully added into centrifuge tube with 1000 μ L pipette tips are adherent；
5) 20min is centrifuged with 2000rpm revolving speed；
6) a little upper layer yellow serum is sopped up using 1000 μ l pipette tips, rear white confluent monolayer cells to the new centrifuge tube of suction (cannot It is drawn onto red blood cell)；
7) isometric phosphate buffer (PBS) is taken to be centrifuged 15min with 1500rpm revolving speed；
8) PBS is outwelled, centrifuge tube is upside down on paper and is absorbed water；
9) 3ml erythrocyte cracked liquid is added in centrifuge tube, cracks 5min after blowing and beating uniformly, during which turns upside down frequently mixed It is even；
10) 5min is centrifuged with 1000rpm revolving speed；
11) erythrocyte cracked liquid is outwelled, centrifuge tube is upside down on paper and is absorbed water；
12) 5ml PBS is added, 5min is centrifuged with 1000rpm revolving speed；
13) PBS is outwelled, white precipitate is peripheral blood mononuclear cells (PBMC).
Step 2, when the human peripheral blood immune cell function fully assess kit in all antibody be liquid When preparation, take 1 μ l-5 μ l (such as 1 μ l, 2 μ l, 3 μ l, 4 μ l, 5 μ l) respectively from all antibody, all with the peripheral blood list A nucleus is mixed, and is protected from light incubation 10 under the conditions of 2 DEG C -6 DEG C (such as 2 DEG C, 3 DEG C, 4 DEG C, 5 DEG C, 6 DEG C, preferably 4 DEG C) Minute to 30 minutes (such as 10 minutes, 15 minutes, 20 minutes, 25 minutes, 30 minutes), obtains cell detection sample；
It is first when at least a kind of antibody is pulvis in human peripheral blood immune cell function comprehensive assessment kit Pulvis is first dissolved as by antibody-solutions using appropriate phosphate buffer (PBS), is divided from the antibody of all liq form later 1 μ l-5 μ l (such as 1 μ l, 2 μ l, 3 μ l, 4 μ l, 5 μ l) is not taken, is all mixed with the peripheral blood mononuclear cells, 2 Incubation 10 minutes to 30 minutes (such as 10 is protected from light under the conditions of DEG C -6 DEG C (such as 2 DEG C, 3 DEG C, 4 DEG C, 5 DEG C, 6 DEG C, preferably 4 DEG C) Minute, 15 minutes, 20 minutes, 25 minutes, 30 minutes), obtain cell detection sample.
Specifically, the condition being protected from light refers to intensity of illumination in 0.2lux environment below.
Specifically, the concentration of antibody is 0.1-2mg/ml in the liquid preparation；The pulvis and phosphate buffer are mixed The concentration of antibody is 0.1-2mg/ml in the antibody-solutions obtained after conjunction.
Step 3 tests and analyzes the cell detection sample using flow cytometer.
In conclusion human peripheral blood immune cell function of the invention, which fully assesses kit, can combine fluidic cell Instrument is for fully assessing the function of the immunocyte in human peripheral blood, easy to use and safe to the human body, the reagent Box can be used for the panimmunities functional assessment such as human immunity health control evaluation and the immune function assessment of infections relating patient In project.Human peripheral blood immune cell function of the invention fully assesses method can be to the immunocyte in human peripheral blood Function fully assessed, operating procedure is simple, safe operation process.
The numberical range of each technological parameter as involved in the present invention can not all embody in the above-described embodiments, As long as but those skilled in the art's envisioned any numerical value fallen into the above-mentioned numberical range completely can be implemented this Invention also includes any combination of occurrence in several numberical ranges certainly.Herein, due to space considerations, be omitted to Out in certain one or more numberical range occurrence embodiment, this disclosure for being not to be construed as technical solution of the present invention do not fill Point.
The Applicant declares that the present invention is explained by the above embodiments detailed process equipment and process flow of the invention, But the present invention is not limited to the above detailed process equipment and process flow, that is, it is above-mentioned detailed not mean that the present invention must rely on Process equipment and process flow could be implemented.It should be clear to those skilled in the art, any improvement in the present invention, Addition, concrete mode selection of equivalence replacement and auxiliary element to each raw material of product of the present invention etc., fall in protection of the invention In range.

Claims (10)

1. a kind of human peripheral blood immune cell function fully assesses kit characterized by comprising with fluorescein-labeled Anti-CD3antibody is marked with fluorescein-labeled anti-human CD4 antibody, with fluorescein-labeled anti-human CD8 antibody, band fluorescein Anti human CD 19 antibody, with fluorescein-labeled anti-human CD21 antibody, with fluorescein-labeled anti-human CD24 antibody, band fluorescein Label anti-human CD25 antibody, with fluorescein-labeled anti-human CD27 antibody, with fluorescein-labeled anti-human CD28 antibody, with glimmering Light element label anti-human CD38 antibody, with fluorescein-labeled anti-human CD56 antibody, with fluorescein-labeled anti-human CD57 antibody, With fluorescein-labeled anti-human CD94 antibody, with fluorescein-labeled anti-human CD127 antibody, with fluorescein-labeled anti-human CD45RA antibody, with fluorescein-labeled anti-human CXCR3 antibody, with fluorescein-labeled anti-human CXCR5 antibody, band fluorescein mark Note anti-human CCR4 antibody, with fluorescein-labeled anti-human CCR6 antibody, with fluorescein-labeled anti-human CCR7 antibody, band fluorescence The anti-human HLA-DR antibodies of element label resist with fluorescein-labeled anti-human PD-1 antibody, with fluorescein-labeled anti-human NKP30 Body, with fluorescein-labeled anti-human NKP46 antibody, with fluorescein-labeled anti-human NKG2D antibody, with fluorescein-labeled anti-human KIR antibody resists with fluorescein-labeled anti-human γ anti-δ, with fluorescein-labeled anti-human 2 antibody of V δ, with fluorescein-labeled People IgD antibody, with fluorescein-labeled anti-human IgM antibodies.
2. human peripheral blood immune cell function as described in claim 1 fully assesses kit, which is characterized in that described anti- It is people CD3 antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, resists anti-human CD27 antibody People CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 It is antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP30 antibody, anti-human It is NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human 2 antibody of V δ, anti-human IgD antibody, anti-human IgM antibody is monoclonal antibody.
3. human peripheral blood immune cell function as described in claim 1 fully assesses kit, which is characterized in that described anti- It is people CD3 antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, resists anti-human CD27 antibody People CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 It is antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP30 antibody, anti-human It is NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human 2 antibody of V δ, anti-human IgD antibody, anti-human Any one in IgM antibody is pulvis or liquid preparation.
4. human peripheral blood immune cell function as claimed in claim 3 fully assesses kit, which is characterized in that described anti- It is people CD3 antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, resists anti-human CD27 antibody People CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 It is antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP30 antibody, anti-human It is NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human 2 antibody of V δ, anti-human IgD antibody, anti-human IgM antibody is all made of the encapsulation of antibody reagent pipe, and the antibody reagent pipe is light transmittance in 10% brown plastic tube below.
5. human peripheral blood immune cell function as claimed in claim 4 fully assesses kit, which is characterized in that described anti- The light transmittance of body Reagent Tube is zero, i.e., completely opaque.
6. human peripheral blood immune cell function as claimed in claim 4 fully assesses kit, which is characterized in that described anti- When the antibody contained in body Reagent Tube is liquid preparation, the volume of antibody is 0.5ml-5ml in the antibody reagent pipe.
7. human peripheral blood immune cell function as claimed in claim 3 fully assesses kit, which is characterized in that described anti- It is people CD3 antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, resists anti-human CD27 antibody People CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 It is antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP30 antibody, anti-human It is NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human 2 antibody of V δ, anti-human IgD antibody, anti-human IgM antibody is liquid preparation.
8. human peripheral blood immune cell function as claimed in claim 3 fully assesses kit, which is characterized in that described anti- It is people CD3 antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, resists anti-human CD27 antibody People CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 It is antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP30 antibody, anti-human It is NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human 2 antibody of V δ, anti-human IgD antibody, anti-human At least one of IgM antibody is pulvis, and it further includes phosphoric acid that the human peripheral blood immune cell function, which fully assesses kit, Salt buffer.
9. human peripheral blood immune cell function as claimed in claim 3 fully assesses kit, which is characterized in that described anti- It is people CD3 antibody, anti-human CD4 antibody, anti-human CD8 antibody, anti human CD 19 antibody, anti-human CD21 antibody, anti-human CD24 antibody, anti-human CD25 antibody, anti-human CD28 antibody, anti-human CD38 antibody, anti-human CD56 antibody, anti-human CD57 antibody, resists anti-human CD27 antibody People CD94 antibody, anti-human CD127 antibody, anti-human CD 45 RA antibody, anti-human CXCR3 antibody, anti-human CXCR5 antibody, anti-human CCR4 It is antibody, anti-human CCR6 antibody, anti-human CCR7 antibody, anti-human HLA-DR antibodies, anti-human PD-1 antibody, anti-human NKP30 antibody, anti-human It is NKP46 antibody, anti-human NKG2D antibody, anti-human KIR antibody, anti-human γ anti-δ, anti-human 2 antibody of V δ, anti-human IgD antibody, anti-human IgM antibody fluorescein label selected from PerCP-Cy5.5, BV510, FITC, AF647, PE-Cy7, BV421, PE, AF647, APC-H7、BB515。
10. a kind of human peripheral blood immune cell function fully assesses method, which comprises the steps of:
Step 1, the human peripheral blood immune cell function provided as described in any one of claim 1 to claim 9 are comprehensive Assess kit；
Peripheral blood mononuclear cells is extracted from 2ml-10ml human peripheral blood；
Step 2, when the human peripheral blood immune cell function fully assess kit in all antibody be liquid preparation When, it takes 1 μ l-5 μ l respectively from all antibody, is all mixed with the peripheral blood mononuclear cells, in 2 DEG C of -6 DEG C of conditions Under be protected from light incubation 10 minutes to 30 minutes, obtain cell detection sample；
When at least a kind of antibody is pulvis in human peripheral blood immune cell function comprehensive assessment kit, adopt first Pulvis is dissolved as antibody-solutions with phosphate buffer, takes 1 μ l-5 μ l respectively from the antibody of all liq form later, entirely Portion is mixed with the peripheral blood mononuclear cells, and incubation 10 minutes to 30 minutes is protected from light under the conditions of 2 DEG C -6 DEG C, is obtained thin Born of the same parents detect sample；
The concentration of antibody is 0.1-2mg/ml in the liquid preparation；The pulvis obtains anti-after mixing with phosphate buffer The concentration of antibody is 0.1-2mg/ml in liquid solution；
Step 3 tests and analyzes the cell detection sample using flow cytometer.
CN201811191491.7A 2018-10-12 2018-10-12 Human peripheral blood immune cell function fully assesses kit and appraisal procedure Pending CN109254147A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201811191491.7A CN109254147A (en) 2018-10-12 2018-10-12 Human peripheral blood immune cell function fully assesses kit and appraisal procedure

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201811191491.7A CN109254147A (en) 2018-10-12 2018-10-12 Human peripheral blood immune cell function fully assesses kit and appraisal procedure

Publications (1)

Publication Number Publication Date
CN109254147A true CN109254147A (en) 2019-01-22

Family

ID=65045197

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201811191491.7A Pending CN109254147A (en) 2018-10-12 2018-10-12 Human peripheral blood immune cell function fully assesses kit and appraisal procedure

Country Status (1)

Country Link
CN (1) CN109254147A (en)

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110441507A (en) * 2019-08-16 2019-11-12 广东普锐生物科技有限公司 A kind of reagent combination, its kit and its method for detecting patients with uterine myoma immune function
CN110488012A (en) * 2019-08-30 2019-11-22 广东普锐生物科技有限公司 A kind of auxiliary diagnostic box of pneumonia of newborn and application
CN110488009A (en) * 2019-04-12 2019-11-22 北京市理化分析测试中心 A kind of detection method and application of the immune function of human body of non-disease diagnostic purpose
CN110687292A (en) * 2019-11-11 2020-01-14 北京大学人民医院(北京大学第二临床医学院) Application of double negative B cell and regulatory B10 cell in preparation of autoimmune disease diagnostic reagent
CN111430034A (en) * 2020-05-08 2020-07-17 广州康途活细胞医疗科技有限责任公司 Human immune system function level detection data model and application thereof in health analysis
CN111579779A (en) * 2020-05-09 2020-08-25 中山大学附属第一医院 Marker for evaluating immune cell function of hepatitis B patient and application thereof
CN112331344A (en) * 2020-11-12 2021-02-05 深圳泛因医学有限公司 Immune state evaluation method and application
WO2021091377A1 (en) * 2019-11-05 2021-05-14 Stichting Euroflow Means and methods for multiparameter cytometry-based leukocyte subsetting.
CN113109575A (en) * 2021-04-16 2021-07-13 浙江普罗亭健康科技有限公司 40 antibody kit for monitoring human immune state and application
CN113125755A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 9 antibody kit for monitoring human immune state and application thereof
CN113125754A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 23 antibody kit for monitoring human immune state and application
CN113125718A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 45 antibody kit for monitoring human immune state and application
CN113125733A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 42 antibody kit for monitoring human immune state and application thereof
CN113433308A (en) * 2021-06-02 2021-09-24 河北森朗泰禾生物科技有限公司 Analysis method of isolated blood sample and immunity evaluation device
CN114112868A (en) * 2020-08-26 2022-03-01 上海睿昂基因科技股份有限公司 Flow cytometry kit for monitoring tumor-related immune microenvironment and monitoring method
CN114791411A (en) * 2022-04-21 2022-07-26 广州先康达生物科技有限公司 Index combination, kit and method for evaluating human immune function
WO2022179620A1 (en) * 2021-02-25 2022-09-01 克莱格医学有限公司 Cd94 engineered cell and composition thereof
CN115166252A (en) * 2022-07-07 2022-10-11 泰州宸安生物科技有限公司 Lymphocyte subset grouping and quantitative detection kit, detection method and application thereof
CN116430055A (en) * 2023-04-21 2023-07-14 中南大学湘雅二医院 T cell immunological state marker combination for evaluating chronic pathological state of T lymphocyte of disease and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107209101A (en) * 2014-10-30 2017-09-26 鹿特丹伊拉斯谟大学医疗中心 For the reagent of diagnosing primary immune deficiency, method and kit
CN108508196A (en) * 2018-04-13 2018-09-07 沈阳汇敏源生物科技有限责任公司 A kind of kit and detection method of detection people's regulatory T cells hypotype

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107209101A (en) * 2014-10-30 2017-09-26 鹿特丹伊拉斯谟大学医疗中心 For the reagent of diagnosing primary immune deficiency, method and kit
CN108508196A (en) * 2018-04-13 2018-09-07 沈阳汇敏源生物科技有限责任公司 A kind of kit and detection method of detection people's regulatory T cells hypotype

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
WING KEUNG CHAN等: "Multiplex and genome-wide analyses reveal distinctive properties of KIR+ and CD56+ T cells in human blood", 《THE JOURNAL OF IMMUNOLOGY》 *
ZICHENG HU等: "MetaCyto: A Tool for Automated Meta-analysis of Mass and Flow Cytometry Data", 《CELL REPORTS》 *

Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110488009A (en) * 2019-04-12 2019-11-22 北京市理化分析测试中心 A kind of detection method and application of the immune function of human body of non-disease diagnostic purpose
CN110488009B (en) * 2019-04-12 2023-09-05 北京市理化分析测试中心 Detection method and application of human immune function for non-disease diagnosis purpose
CN110441507A (en) * 2019-08-16 2019-11-12 广东普锐生物科技有限公司 A kind of reagent combination, its kit and its method for detecting patients with uterine myoma immune function
CN110488012A (en) * 2019-08-30 2019-11-22 广东普锐生物科技有限公司 A kind of auxiliary diagnostic box of pneumonia of newborn and application
CN110488012B (en) * 2019-08-30 2023-02-03 广东普锐生物科技有限公司 Auxiliary diagnosis kit for neonatal pneumonia and application
WO2021091377A1 (en) * 2019-11-05 2021-05-14 Stichting Euroflow Means and methods for multiparameter cytometry-based leukocyte subsetting.
CN110687292A (en) * 2019-11-11 2020-01-14 北京大学人民医院(北京大学第二临床医学院) Application of double negative B cell and regulatory B10 cell in preparation of autoimmune disease diagnostic reagent
CN111430034A (en) * 2020-05-08 2020-07-17 广州康途活细胞医疗科技有限责任公司 Human immune system function level detection data model and application thereof in health analysis
CN111579779A (en) * 2020-05-09 2020-08-25 中山大学附属第一医院 Marker for evaluating immune cell function of hepatitis B patient and application thereof
CN114112868A (en) * 2020-08-26 2022-03-01 上海睿昂基因科技股份有限公司 Flow cytometry kit for monitoring tumor-related immune microenvironment and monitoring method
CN112331344B (en) * 2020-11-12 2021-08-03 深圳泛因医学有限公司 Immune state evaluation method and application
CN112331344A (en) * 2020-11-12 2021-02-05 深圳泛因医学有限公司 Immune state evaluation method and application
WO2022179620A1 (en) * 2021-02-25 2022-09-01 克莱格医学有限公司 Cd94 engineered cell and composition thereof
CN113109575A (en) * 2021-04-16 2021-07-13 浙江普罗亭健康科技有限公司 40 antibody kit for monitoring human immune state and application
CN113125755A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 9 antibody kit for monitoring human immune state and application thereof
CN113125733A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 42 antibody kit for monitoring human immune state and application thereof
CN113125754A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 23 antibody kit for monitoring human immune state and application
CN113125718A (en) * 2021-04-16 2021-07-16 浙江普罗亭健康科技有限公司 45 antibody kit for monitoring human immune state and application
CN113433308A (en) * 2021-06-02 2021-09-24 河北森朗泰禾生物科技有限公司 Analysis method of isolated blood sample and immunity evaluation device
CN114791411A (en) * 2022-04-21 2022-07-26 广州先康达生物科技有限公司 Index combination, kit and method for evaluating human immune function
CN115166252A (en) * 2022-07-07 2022-10-11 泰州宸安生物科技有限公司 Lymphocyte subset grouping and quantitative detection kit, detection method and application thereof
CN116430055A (en) * 2023-04-21 2023-07-14 中南大学湘雅二医院 T cell immunological state marker combination for evaluating chronic pathological state of T lymphocyte of disease and application thereof
CN116430055B (en) * 2023-04-21 2024-07-30 中南大学湘雅二医院 T cell immunological state marker combination for evaluating chronic pathological state of T lymphocyte of disease and application thereof

Similar Documents

Publication Publication Date Title
CN109254147A (en) Human peripheral blood immune cell function fully assesses kit and appraisal procedure
CN109406775A (en) Autoimmune disease patient's immune function assesses kit and appraisal procedure
Kroy et al. Liver environment and HCV replication affect human T-cell phenotype and expression of inhibitory receptors
Sauce et al. Lymphopenia-driven homeostatic regulation of naive T cells in elderly and thymectomized young adults
CN109212215A (en) Tumor patient immune cell function assesses kit and appraisal procedure
Hausman et al. Defective monocyte function in patients with genitourinary carcinoma
Tjio et al. Production of human immunoglobulin E antibody in vitro
Nikolajczyk et al. The outliers become a stampede as immunometabolism reaches a tipping point
Kawakami et al. Reduced immune function and malnutrition in the elderly
US20080081076A1 (en) Nanofraction immune modulators, preparations and compositions including the same, and associated methods
El-Badawy et al. Relations of regulatory T cells with hepatitis markers in chronic hepatitis B virus infection
CN109254148A (en) Human peripheral blood T cell immune function simplifies assessment kit and appraisal procedure
Mustafa et al. T lymphocyte subpopulations in anorexia nervosa and refeeding
Dong et al. Isolation of murine hepatic lymphocytes using mechanical dissection for phenotypic and functional analysis of NK1. 1+ cells
CN114791411A (en) Index combination, kit and method for evaluating human immune function
McClure et al. Evaluation of immune parameters in propanil-exposed farm families
Zhang et al. IL-15 induced bystander activation of CD8+ T cells may mediate endothelium injury through NKG2D in Hantaan virus infection
Broder et al. Current concepts in immunoregulatory T-cell neoplasms
Siewe et al. A role for TLR signaling during B cell activation in antiretroviral-treated HIV individuals
Chou et al. Erythrocyte-bound immune complexes trigger the release of interleukin-1 from human monocytes
CN102755633B (en) Application of human recombinant protein PDCD5 to treatment of autoimmune disease
Yang et al. Effector Memory CD8+ and CD4+ T Cell Immunity Associated with Metabolic Syndrome in Obese Children
Iyengar et al. Temporal expression of cytokines and B-cell phenotypes during mechanical circulatory support
CN109803681A (en) For treating the immunosuppressant composite of immune disorders
Gmyrek et al. Differential flow cytometric detection of intracellular cytokines in peripheral and peritoneal mononuclear cells of women with endometriosis

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication
RJ01 Rejection of invention patent application after publication

Application publication date: 20190122