Antibody composition for detecting abnormal CTL (cytotoxic T lymphocyte) cells of hemophagocytic syndrome and application of antibody composition

Abstract

The invention relates to an antibody composition for detecting abnormal CTL (cytotoxic T lymphocyte) cells of hemophagocytic syndrome and application of the antibody composition, and the antibody composition comprises a CD2 antibody, a CD3 antibody, a CD4 antibody, a CD5 antibody, a CD7 antibody, a CD8 antibody, a CD56 antibody, a CD57 antibody, a Granzyme B antibody, a Perforin antibody, a TRBC1 antibody and a CD45 antibody. In the technical scheme provided by the invention, key phenotypic characteristics of CTL (Cytotoxic Lymphocyte) can be completely covered by various antibodies aiming at T cell and NK cell surface markers, T large granular lymphocyte markers, cytotoxic granular proteins, T cell receptor constant regions and leukocyte common antigens; wherein the clonality of the CTL cells is judged through the expression level of the TRBC1 antibody, and the cytotoxic function activity of the CTL cells is evaluated through the expression levels of Granzyme B and Perforin.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biotechnology, in particular to an antibody composition for detecting abnormal CTL cells in hemophagocytic syndrome and application thereof. BACKGROUND

[0002] Hemophagocytic syndrome (HLH) is closely related to abnormal cytotoxic T lymphocyte (CTL) function. Under normal circumstances, CTL cells induce the programmed death (apoptosis) of target cells in the immune synapse by activating the perforin-granzyme cell death pathway, thereby clearing antigens. However, in HLH, due to genetic defects and other reasons, the killing function of CTL cells is impaired, and antigens cannot be effectively cleared, leading to persistent stimulation of the immune system. Genetic defects also impair the function of CTL cell lysosome-associated membrane glycoprotein CD107a to transfer to the cell surface, thereby affecting the degranulation process and failing to normally release cytotoxic granules such as perforin and granzyme. This dysfunction can lead to an inability to normally terminate the immune response, triggering excessive inflammatory reactions. CTL cell function defects can also cause abnormal expansion of antigen-specific effector T cells: on the one hand, CD8+ T cells cannot clear antigen-presenting cells; on the other hand, cytotoxic cells lack the function of down-regulating immune responses. This causes activated lymphocytes to secrete large amounts of gamma-interferon, further activating macrophages, which in turn promote more T cell activation, forming a vicious cycle. Therefore, screening abnormal CTL cells in the bone marrow and peripheral blood of patients with hemophagocytic syndrome is a very important detection index in the process of disease diagnosis and follow-up.

[0003] Most of the existing detection groups are for detecting T cell immunophenotypes, without detecting functional indicators such as granzyme and perforin. SUMMARY

[0004] Based on the above description, the present application provides an antibody composition for detecting abnormal CTL cells in hemophagocytic syndrome and application thereof, aiming to solve the problem of how to screen abnormal CTL cells in HLH patients.

[0005] The technical solution of the present application to solve the above technical problems is as follows: The present application provides an antibody composition for detecting abnormal CTL cells in hemophagocytic syndrome, comprising CD2 antibody, CD3 antibody, CD4 antibody, CD5 antibody, CD7 antibody, CD8 antibody, CD56 antibody, CD57 antibody, Granzyme B antibody, Perforin antibody, TRBC1 antibody and CD45 antibody.

[0006] Further, the volume ratio of the CD2 antibody, the CD3 antibody, the CD4 antibody, the CD5 antibody, the CD7 antibody, the CD8 antibody, the CD56 antibody, the CD57 antibody, the Granzyme B antibody, the Perforin antibody, the TRBC1 antibody, the CD45 antibody is 2:2:2:1:1:2:2:2:2:2:1:1.

[0007] Further, each antibody is a fluorescein-labeled antibody.

[0008] Further, the fluorescein label of the CD2 antibody is AlexaFluor 700; the fluorescein label of the CD3 antibody is Brilliant Violet 750; the fluorescein label of the CD4 antibody is PE-Cy7; the fluorescein label of the CD5 antibody is APC; the fluorescein label of the CD7 antibody is BV421; the fluorescein label of the CD8 antibody is APC-CY7; the fluorescein label of the CD56 antibody is ECD; the fluorescein label of the CD57 antibody is Brilliant Violet 605; the fluorescein label of the Granzyme B antibody is PE; the fluorescein label of the Perforin antibody is PerCP-Cy5.5; the fluorescein label of the TRBC1 antibody is FITC; the fluorescein label of the CD45 antibody is QB500.

[0009] The application also provides a kit for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells, comprising the antibody composition for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells as described above.

[0010] Further, the kit further comprises a PBS buffer, a red blood cell lysis solution and a membrane breaker.

[0011] Further, the concentration of the PBS buffer is 0.02 mol / L, and the pH value is 7.2-7.6.

[0012] The application also provides a use of the antibody composition for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells as described above or the kit for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells as described above in detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells.

[0013] Further, the detecting the abnormal CTL cell of the hemophagocytic syndrome comprises the following steps: S1. obtaining a sample to be tested, mixing CD2 antibody, CD3 antibody, CD4 antibody, CD5 antibody, CD7 antibody, CD8 antibody, CD56 antibody, CD57 antibody, TRBC1 antibody, CD45 antibody with the sample to be tested, avoiding light incubation, then adding red blood cell lysis solution, avoiding light incubation, centrifuging, removing supernatant, and obtaining a lysis sample; S2. mixing the lysis sample with PBS buffer, washing, centrifuging, removing supernatant, adding Granzyme B antibody, Perforin antibody and membrane breaker for incubation, adding PBS buffer, washing, centrifuging, removing supernatant, and obtaining a membrane breaking sample; S3. resuspending the membrane breaking sample with PBS buffer, placing on a flow cytometer for detection and analysis, judging the clonality of the CTL cell through TRBC1, and judging the cytotoxic function thereof through Granzyme B and Perforin; The CD2 antibody, the CD3 antibody, the CD4 antibody, the CD5 antibody, the CD7 antibody, the CD8 antibody, the CD56 antibody, the CD57 antibody, the Granzyme B antibody, the Perforin antibody, the TRBC1 antibody and the CD45 antibody are the antibody composition for detecting the abnormal CTL cell of the hemophagocytic syndrome as described above.

[0014] Further, in step S1, the concentration of the sample to be tested is 4×10 6 ~ 5×10 6 cells / mL.

[0015] Compared with the prior art, the technical scheme has the following beneficial technical effects: In the technical scheme, a plurality of antibodies for T cell and NK cell surface markers (CD2, CD3, CD4, CD5, CD7, CD8, CD56), T large granular lymphocyte markers (CD57), cytotoxic granule proteins (Granzyme B, Perforin), T cell receptor constant region (TRBC1) and white blood cell common antigen (CD45) can comprehensively cover the key phenotypic characteristics of CTL (cytotoxic T lymphocyte), and the recognition ability of the abnormal CTL cell in the hemophagocytic syndrome is significantly improved through the joint detection of multi-parameter flow cytometry, the normal CTL and pathological CTL are effectively distinguished, and the sensitivity and specificity of the detection are improved. The clonality of the CTL cell is judged through the expression level of the TRBC1 antibody, and the cytotoxic function activity thereof is evaluated through the expression levels of Granzyme B and Perforin. Attached Figure Description

[0016] Figure 1 The flow cytometry analysis of the sample is shown in Example 2 of this application, which is the method for detecting abnormal CTL cells of hemophagocytic cell syndrome. Detailed Implementation

[0017] To facilitate understanding of this application, a more complete description will be provided below with reference to the accompanying drawings, which illustrate embodiments of the present application. However, the present application can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that the disclosure of this application will be thorough and complete.

[0018] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0019] HLH is closely related to abnormal CTL cell function. Under normal circumstances, CTL cells induce programmed cell death (apoptosis) of target cells in the immune synapse by activating the perforin-granzyme cell death pathway, thereby clearing antigens. However, in HLH, due to genetic defects and other reasons, the killing function of CTL cells is impaired, failing to effectively clear antigens, leading to persistent antigen stimulation of the immune system. Gene defects also impair the function of CTL cell lysosome-associated membrane glycoprotein CD107a in translocating to the cell surface, thus affecting its degranulation process and preventing the normal release of cytotoxic particles such as perforin and granzyme. This dysfunction can lead to the inability to terminate the immune response normally, triggering an excessive inflammatory response. CTL cell dysfunction may also lead to abnormal proliferation of antigen-specific effector T cells: on the one hand, CD8+ T cells are unable to clear antigen-presenting cells; on the other hand, the downregulation function of cytotoxic cells in the immune response is lost. This causes activated lymphocytes to secrete large amounts of gamma interferon, further activating macrophages, which in turn promote the activation of more T cells, forming a vicious cycle. Therefore, screening for abnormal CTL cells in the bone marrow and peripheral blood of patients with hemophagocytic lymphohistiocytosis (HLH) is a very important indicator in the diagnosis and follow-up of the disease.

[0020] Most existing detection kits are designed to detect T-cell immunophenotypes, but do not detect functional indicators such as granzymes and perforin.

[0021] In view of this, the application provides an antibody composition for detecting abnormal CTL cells in hemophagocytic syndrome, comprising CD2 antibody, CD3 antibody, CD4 antibody, CD5 antibody, CD7 antibody, CD8 antibody, CD56 antibody, CD57 antibody, Granzyme B antibody, Perforin antibody, TRBC1 antibody and CD45 antibody.

[0022] In the technical scheme of the application, a plurality of antibodies for T cell and NK cell surface markers (CD2, CD3, CD4, CD5, CD7, CD8, CD56), T large granular lymphocyte markers (CD57), cytotoxic granule proteins (Granzyme B, Perforin), T cell receptor constant region (TRBC1) and leukocyte common antigen (CD45) can comprehensively cover the key phenotypic characteristics of CTL (cytotoxic T lymphocyte), and the recognition ability of abnormal CTL cells in hemophagocytic syndrome is significantly improved through joint detection by multi-parameter flow cytometry, effectively distinguishing normal CTL from pathological CTL, and improving the sensitivity and specificity of detection.

[0023] Further, the volume ratio of the CD2 antibody, the CD3 antibody, the CD4 antibody, the CD5 antibody, the CD7 antibody, the CD8 antibody, the CD56 antibody, the CD57 antibody, the Granzyme B antibody, the Perforin antibody, the TRBC1 antibody and the CD45 antibody is 2:2:2:1:1:2:2:2:2:2:1:1.

[0024] In the technical scheme of the application, by accurately regulating the volume ratio of each antibody, the best balance of signal intensity, specificity and sensitivity in multi-color flow detection is achieved, not only improving the recognition ability of abnormal CTL cells in hemophagocytic syndrome, but also enhancing the stability, repeatability and clinical applicability of detection; wherein CD45 as a general marker for leukocytes, its appropriate proportion (1 part) can clearly distinguish the lymphocyte group and exclude debris and non-leukocyte interference, CD3 and CD8 are in equal amount (2:2) to accurately define the main group of CTL; CD4 (2 parts) and CD8 are used to distinguish double positive or abnormal phenotype cells; CD56 and CD57 (each 2 parts) are used to identify NK-like and T large granular lymphocyte subgroups; TRBC1 (1 part) is used for clonality analysis to avoid false positives caused by excessive staining.

[0025] Further, each antibody is a fluorescein-labeled antibody.

[0026] In the technical solution of the application, all the antibodies are labeled with fluorescein, multiple target molecules can be simultaneously recognized in a single flow cytometry detection, and multi-dimensional combined analysis of T cell surface markers, cytotoxic function and clonal characteristics is realized. Compared with non-fluorescent labeling or detection method, the detection time is greatly shortened, and the experimental throughput is improved.

[0027] Further, the fluorescein labeling of the CD2 antibody is AlexaFluor 700; The fluorescein labeling of the CD3 antibody is BrilliantViolet750; The fluorescein labeling of the CD4 antibody is PE-Cy7; The fluorescein labeling of the CD5 antibody is APC; The fluorescein labeling of the CD7 antibody is BV421; The fluorescein labeling of the CD8 antibody is APC-CY7; The fluorescein labeling of the CD56 antibody is ECD; The fluorescein labeling of the CD57 antibody is BrilliantViolet605; The fluorescein labeling of the Granzyme B antibody is PE; The fluorescein labeling of the Perforin antibody is PerCP-Cy5.5; The fluorescein labeling of the TRBC1 antibody is FITC; The fluorescein labeling of the CD45 antibody is QB500.

[0028] In the technical solution of the application, by designing the type of fluorescein labeling of each antibody, the optimal allocation of spectral resources, the accurate balance of signal strength and the high compatibility of the detection system are realized, not only the technical advantages of multi-parameter flow cytometry are fully utilized, but also the synergistic effect of antibody types and proportions is formed, a sensitive, accurate and robust abnormal CTL detection system is constructed, which has outstanding technical innovation and important clinical transformation value.

[0029] The application further provides a kit for detecting hemophagocytic syndrome abnormal CTL cells, comprising the antibody composition for detecting hemophagocytic syndrome abnormal CTL cells as described above.

[0030] Since the kit adopts all the technical solutions of the above embodiments, it at least has all the beneficial effects brought by the technical solutions of the above embodiments, which will not be repeated here.

[0031] Further, the kit further comprises PBS buffer, red blood cell lysis solution and membrane breaker.

[0032] In the technical solution of this invention, PBS buffer is used for sample washing and dilution, maintaining cell physiological state and antibody binding activity; red blood cell lysis buffer can quickly and efficiently remove red blood cell interference from whole blood or bone marrow samples, avoiding its influence on lymphocyte gating and fluorescence signal detection; and the membrane permeability agent is used for cell membrane permeability treatment, allowing fluorescently labeled antibodies to enter the cell and specifically recognize intracellular proteins such as Granzyme B and Perforin. The three components work synergistically to ensure that the entire process from sample preparation to surface / intracellular double staining is completed under uniform and optimized conditions.

[0033] Furthermore, the concentration of the PBS buffer is 0.02 mol / L, and the pH value is 7.2–7.6.

[0034] In the technical solution of this invention, the sample to be tested can be diluted and washed in a mild environment under PBS conditions of 0.02 mol / L and pH 7.2-7.6, which maximizes the retention of the number of target lymphocytes. It is especially beneficial for the enrichment and detection of low-abundance abnormal CTL cells. It is suitable for automated liquid handling systems, facilitates high-throughput and standardized operation, and meets the needs of rapid screening and long-term follow-up monitoring in clinical emergency departments.

[0035] The present invention also proposes the application of an antibody composition for detecting hemophagocytic lymphohistiocytic syndrome (CTL) abnormal cells as described above, or a kit for detecting hemophagocytic lymphohistiocytic syndrome (CTL) abnormal cells as described above, in the detection of hemophagocytic lymphohistiocytic syndrome (CTL) abnormal cells.

[0036] Furthermore, the detection of hemophagocytic lymphohistiocytosis syndrome-abnormal CTL cells includes the following steps: S1. Obtain the sample to be tested, mix CD2 antibody, CD3 antibody, CD4 antibody, CD5 antibody, CD7 antibody, CD8 antibody, CD56 antibody, CD57 antibody, TRBC1 antibody, and CD45 antibody with the sample to be tested, incubate in the dark, add red blood cell lysis buffer, incubate in the dark, centrifuge, remove the supernatant, and obtain the lysed sample; S2. Mix the lysed sample with PBS buffer, wash, centrifuge, remove the supernatant, add Granzyme B antibody, Perforin antibody and membrane perforation agent for incubation, add PBS buffer, wash, centrifuge, remove the supernatant to obtain the membrane perforated sample; S3. The perforated sample was mixed with PBS buffer and resuspended, then placed on a flow cytometer for detection and analysis. TRBC1 was used to determine the clonality of CTL cells, and Granzyme B and Perforin were used to determine their cytotoxic function. The CD2 antibody, the CD3 antibody, the CD4 antibody, the CD5 antibody, the CD7 antibody, the CD8 antibody, the CD56 antibody, the CD57 antibody, the Granzyme B antibody, the Perforin antibody, the TRBC1 antibody and the CD45 antibody are the antibody composition for detecting abnormal CTL cells of hemophagocytic syndrome as described above.

[0037] In the technical solution of the present application, the standard process of surface staining, lysis and intracellular staining is provided to ensure that the surface markers are fully combined with the antibodies in the complete cell membrane environment, and the influence of the membrane protein structure by the membrane breaking process is avoided; then the Granzyme B and Perforin antibodies are effectively entered into the cell to complete the staining by the membrane breaking agent, the high specificity recognition of all targets is realized, the immunophenotype and functional state information of the abnormal CTL are comprehensively obtained, and the multi-dimensional diagnostic basis is provided for the clinic.

[0038] Further, in step S1, the concentration of the sample to be tested is 4×10 6 ~ 5×10 6 cells / mL.

[0039] In the technical solution of the present application, the sample concentration is controlled in the range of 4×10 6 ~ 5×10 6 cells / mL, which ensures that the cell density in unit volume is moderate, avoids the antibody binding site competition, insufficient staining or local aggregation phenomenon caused by too high cell density, and prevents the statistical error and rare cell group missing caused by too few cells.

[0040] The technical solution of the present application will be further described in detail in combination with specific examples, and it should be understood that the following examples are only used to explain the present application, and are not used to limit the present application.

[0041] The materials, reagents and the like used in the following examples can be obtained from commercial channels unless otherwise specified, and the main reagents are shown in Table 1.

[0042] Table 1 Main reagents

[0043] Example 1 The antibody composition provided in this example is shown in Table 2.

[0044] Table 2 Components and contents of the antibody composition

[0045] Example 2 The embodiment provides a method for detecting abnormal CTL cells of hemophagocytic syndrome, comprising the following steps: (1) Adjust the concentration of the human bone marrow sample to be detected to 4x10 6 ~ 5x10 6 According to the amount in Table 1, add AlexaFluor 700-labeled CD2 antibody, BrilliantViolet750-labeled CD3 antibody, PE-Cy7-labeled CD4 antibody, APC-labeled CD5 antibody, BrilliantViolet 421-labeled CD7 antibody, APC-CY7-labeled CD8 antibody, ECD-labeled CD56 antibody, BrilliantViolet605-labeled CD57 antibody, FITC-labeled TRBC1 antibody, QB500-labeled CD45 antibody into a flow tube in sequence, and add the sample to be detected, so that the number of cells in the flow tube is 3-4x10 5 Add 1-2 mL 1xLysingsolutionforFACS10x red blood cell lysis solution to the flow tube, mix gently, and incubate at room temperature for 10 min, then place the flow tube on a centrifuge at a speed of 400 rcf / min for 5 min, and remove the supernatant.

[0046] (2) Add 1 mL PBS buffer to the flow tube, mix, then centrifuge at a speed of 400 rcf / min for 5 min, remove the supernatant, then add 100 uL of A liquid in FixPERM Kit, fix for 15 min, then add 100 uL of B liquid in FixPERM Kit and PE-labeled Granzyme B antibody and PerCP-Cy5.5-labeled Perforin antibody, and incubate for 20 min, add 1 mL PBS buffer to the flow tube, mix, then centrifuge at a speed of 400 rcf / min for 5 min, remove the supernatant, and resuspend with 150-300 uL PBS buffer.

[0047] (3) Place the flow tube on a flow cytometer for detection, and obtain 5-10 million events, then export the data in fcs format, and analyze the data by using a flow analysis software. Circle the cell population with abnormal pan T antigen expression, judge the clonality thereof by TRBC1, and judge the cytotoxic function thereof by Granzyme B and Perforin.

[0048] The normal T cell phenotype is CD3+CD5+CD2+CD7+, and the normal T cell comprises CD4+CD8-Th cell and CD8+CD4-CTL cell, the TRBC1 expression rate of which is 15%-85%, the Granzyme B expression rate of the normal CTL cell is 15.7%-65.9%, and the Perforin expression rate is 11.0%-64.4%. The normal T cell can partially express CD57, and the large granular lymphocyte fully expresses CD57.

[0049] The results are shown in the following table: Figure 1 As shown in the following table, 22% of CD3+CD5- / dim cells (red in the figure) can be seen in the bone marrow of the patient, express CD2, CD7, CD8, Granzyme B (99.73%), TRBC1 (99.0%), partially express CD56, do not express CD4, CD57 and Perforin (0.35%), the group of cells express CD8 and do not express CD4, which indicates that the property is CTL cell, and the expression of CD5, Granzyme B, TRBC1 and Perforin is abnormal, which indicates that the CTL cell is abnormal CTL cell.

[0050] The above merely describes preferred embodiments of the present application and is not intended to limit the present application, and any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

[0051] In summary, the technical scheme of the present application has the following beneficial technical effects: In the technical scheme of the present application, a plurality of antibodies of T cell and NK cell surface markers (CD2, CD3, CD4, CD5, CD7, CD8, CD56), T large granular lymphocyte markers (CD57), cytotoxic granule proteins (Granzyme B, Perforin), T cell receptor constant region (TRBC1) and white blood cell common antigen (CD45) can comprehensively cover the key phenotype characteristics of CTL (cytotoxic T lymphocyte), and the recognition ability of abnormal CTL cell in hemophagocytic syndrome is significantly improved through joint detection of multi-parameter flow cytometry, the normal CTL and pathological CTL are effectively distinguished, and the sensitivity and specificity of detection are improved; wherein, the expression level of the TRBC1 antibody is used to judge the clonality of the CTL cell, and the expression levels of Granzyme B and Perforin are used to evaluate the cytotoxic function activity.

Claims

1. An antibody composition for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells, characterized by comprising an antibody which binds to a CTLA-4 molecule of a CTL cell. CD2 antibody, CD3 antibody, CD4 antibody, CD5 antibody, CD7 antibody, CD8 antibody, CD56 antibody, CD57 antibody, Granzyme B antibody, Perforin antibody, TRBC1 antibody and CD45 antibody.

2. The antibody composition of claim 1, wherein The volume ratio of the CD2 antibody, the CD3 antibody, the CD4 antibody, the CD5 antibody, the CD7 antibody, the CD8 antibody, the CD56 antibody, the CD57 antibody, the Granzyme B antibody, the Perforin antibody, TRBC1 antibody and the CD45 antibody is 2:2:2:1:1:2:2:2:2:2:1:

1.

3. The antibody composition of claim 1, wherein Each antibody is a fluorescein-labeled antibody.

4. The antibody composition of claim 3, wherein, The fluorescein label of the CD2 antibody is AlexaFluor 700; The fluorescein label of the CD3 antibody is BrilliantViolet750; The fluorescein label of the CD4 antibody is PE-Cy7; The fluorescein label of the CD5 antibody is APC; The fluorescein label of the CD7 antibody is BV421; The fluorescein label of the CD8 antibody is APC-CY7; The fluorescein label of the CD56 antibody is ECD; The fluorescein label of the CD57 antibody is BrilliantViolet605; The fluorescein label of the Granzyme B antibody is PE; The fluorescein label of the Perforin antibody is PerCP-Cy5.5; The fluorescein label of the TRBC1 antibody is FITC; The fluorescein label of the CD45 antibody is QB500.

5. A kit for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells, characterized by, The antibody composition for detecting abnormal CTL cells of hemophagocytic syndrome according to any one of claims 1 to 4.

6. The kit for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells according to claim 5, characterized by, The kit further comprises a PBS buffer, a red blood cell lysing solution and a membrane disrupter.

7. The kit for detecting paroxysmal nocturnal hemoglobinuria abnormal CTL cells according to claim 6, characterized by, The concentration of the PBS buffer is 0.02 mol / L, and the pH value is 7.2-7.

6.

8. Use of the antibody composition for detecting abnormal CTL cells of hemophagocytic syndrome according to any one of claims 1 to 4 or the kit for detecting abnormal CTL cells of hemophagocytic syndrome according to any one of claims 5 to 7 in detecting abnormal CTL cells of hemophagocytic syndrome.

9. Use according to claim 8, characterized in that, The detection of abnormal CTL cells of hemophagocytic syndrome comprises the following steps: S1. Obtain a sample to be tested, mix CD2 antibody, CD3 antibody, CD4 antibody, CD5 antibody, CD7 antibody, CD8 antibody, CD56 antibody, CD57 antibody, TRBC1 antibody, CD45 antibody with the sample to be tested, incubate in the dark, then add a red blood cell lysing solution, incubate in the dark, centrifuge, remove the supernatant, and obtain a lysed sample; S2. Mix the lysed sample with a PBS buffer, wash, centrifuge, remove the supernatant, add Granzyme B antibody, Perforin antibody and a membrane disrupter, incubate, add a PBS buffer, wash, centrifuge, remove the supernatant, and obtain a membrane-disrupted sample; S3. The film-breaking sample is mixed with PBS buffer for resuspension, placed on a flow cytometer for detection and analysis, and the clonality of the sample to be tested is determined by TRBC1, and the function of the sample to be tested is determined by Granzyme B and Perforin; The CD2 antibody, the CD3 antibody, the CD4 antibody, the CD5 antibody, the CD7 antibody, the CD8 antibody, the CD56 antibody, the CD57 antibody, the Granzyme B antibody, the Perforin antibody, the TRBC1 antibody, and the CD45 antibody are the antibody composition for detecting abnormal CTL cells of hemophagocytosis syndrome according to any one of claims 1 to 4.

10. Use according to claim 9, characterized in that, In step S1, the concentration of the sample to be measured is 4 x 10 6 ~ 5 x 10 6 cells / mL.

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