32 results about "Lymphocyte subsets" patented technology

The invention discloses an expansion method of various lymphocyte subpopulations and application of the expansion method to preparation of an adoptive immunotherapy medicine for cancers. The expansion method comprises the following steps: S1, adding IFN gamma and zoledronic acid into a culture medium of PBMC collected in the blood of a tumor patient, and re-adding an OKT3 antibody and recombinant human interleukin-2 after culture to stimulate PBMC so as to finish preliminary expansion; S2, after finish of preliminary expansion, mixing immune lymphocytes with feeder layer cells, and after adding zoledronic acid, an OKT3 antibody and recombinant human interleukin-2, continually performing expansion. By adoption of the expansion method, after twice expansion, the sum of the immune cells can be 10,000 to 80,000 times by expansion, the expanded immune cells include alpha beta T cells, gamma delta T cells, NKT cells and NK cells, and tumor cells can be directly killed, or a cellular immunotherapy drug is prepared to kill the tumor cells.

Leptin was previously demonstrated to exert potent immune modulatory properties in several immune mediated disorders. The aim of the study was to determine leptin's anti-tumor effect in a murine model of human hepatocellular carcinoma (HCC). In vivo, Athymic T cell deficient (nude) mice transplanted with 1×106 human Hep3B cells, followed by administration of two daily intraperitoneal doses of 0.5 mg / gram leptin for 6 weeks. Leptin administration induced a significant reduction in tumor size and improved survival in nude mice. Histologically, tumors of leptin-administered mice featured increased inflammatory exudate in interphase areas. Leptin-induced tumor suppression was associated with a significant increase in peripheral natural killer (NK) cell number. Splenocytes from leptin-treated mice featured decreased expression of CIS mRNA. To determine which lymphocyte subset is a prerequisite for the anti tumor effect of leptin, T&B cell deficient (Scid) mice and T,B& NK deficient (Scid-Beige) mice were subcutaneously implanted with Hep3B tumor cells, with and without the daily intraperitoneal administration of 0.5 mg / gram leptin for 6 weeks. SCID mice featured leptin-associated tumor suppression similar to those of nude mice. In contrast, NK-deficient SCID-Beige mice developed larger tumors. To further establish natural killer cell's central role in mediation of leptin's anti-tumor effect, NK cells were incubated in vitro with increasing doses of leptin, demonstrating a dose-dependent increase in cytotoxic activity. Incubation of leptin with hepatoma cell line was found to induce a dose-dependent reduction in hepatoma cell proliferation, suggesting an additive direct anti-tumor effect. Further synergism in inhibition of hepatoma cell proliferation in vitro was achieved following addition of natural killer cells. HCC cells expressed leptin receptor mRNA, while addition of leptin induced increased mRMA expression of STAT2 and SOCS1 on tumor cell lines. Leptin administration induces a significant suppression of human HCC. This effect is mediated by induction of natural killer cell proliferation and activation, and by direct inhibition of tumor growth. Decreased natural killer cell expression of inhibitory CIS protein and over expression of the anti-proliferative STAT2 and SOCS1 proteins in HCC lines may underline both anti cancerous effects of leptin.

The invention discloses a detection method of human peripheral blood lymphocytes. The preparation method comprises the following steps of: taking human anticoagulation peripheral blood; adding anti-human CD3, CD4, CD8 and CD45 monoclonal antibodies and anti-human CD3, CD56, CD16 and CD45 monoclonal antibodies, then adding a hemolysin working solution containing a fluorescent probe to respectivelyobtain the percentage and absolute count value of a human peripheral blood T lymphocyte subset and an NK lymphocyte subset and achieve the detection of human peripheral blood lymphocytes. The method has the advantages of simplicity and convenience in operation, long sample stable preservation time, high accuracy, multi-index output and the like.

The invention provides a detection method of immune functions of mesenchymal stem cells. The detection method of the immune functions of the mesenchymal stem cells comprises the following steps: (1) co-culturing umbilical cord mesenchymal stem cells and peripheral blood mononuclear cells so as to obtain a sample to be tested; and (2) detecting proliferative activity of lymphocytes and proportion of lymphocyte subsets in the sample to be tested. The detection method of the immune functions of the mesenchymal stem cells is suitable for non-diagnostic purposes. According to the detection method of the immune functions of the mesenchymal stem cells provided by the embodiment of the invention, the mesenchyma stem cells and the human peripheral blood mononuclear cells are co-cultured so as to obtain the sample to be tested; and then, the proportion of the lymphocyte subsets and the proliferative activity of the lymphocytes in the sample to be tested are detected so as to evaluate the immunoregulatory capacity of the mesenchymal stem cells. The greater the changes of the proportion of the lymphocyte subsets and the proliferative activity of the lymphocytes are, the stronger the immune functions and the better the quality of the mesenchymal stem cells are.

The invention discloses a composition for autologous lymphocyte culture, a culture solution and application of the composition. The active ingredients of the composition for autologous lymphocyte culture are composed of plasma, an anti-human CD3 monoclonal antibody, IL-2, IL-7, IL-15 and PHA-P. Compared with a composition for lymphocyte culture as a control, the content of a T lymphocyte subset obtained by using a culture medium containing the composition for induced culture of peripheral blood mononuclear cells is significantly increased; the number of lymphocytes is also significantly increased after 14 days of culture, and the number is increased by 1.15 times; and the killing activity of the lymphocytes on K562 cells is also significantly enhanced, and the killing activity is enhancedby 1.6 times. The composition for autologous lymphocyte culture can be used for specific induced amplification of mononuclear cells to produce autologous lymphocytes with high purity and strong biological activity.

The invention provides a detection method of a decidual lymphocyte subset. The detection method comprises the following steps: carrying out fluorescent staining on the same decidual lymphocyte subset sample by virtue of a fluorescently-labeled anti-CD3 antibody, a fluorescently-labeled anti-CD4 antibody, a fluorescently-labeled anti-CD8 antibody, a fluorescently-labeled anti-CD56 antibody, a fluorescently-labeled anti-CD16 antibody, a fluorescently-labeled anti-IFN-gamma antibody, a fluorescently-labeled anti-IL-4 antibody and Fixable Viability Dye, so as to obtain the stained sample; detecting the stained sample by virtue of a flow cytometry, so as to obtain detection data; and analyzing the detection data. The detection method has the beneficial effects that fewer decidual lymphocyte samples are required, antibodies are saved, and the operation is simple. Besides, the invention further provides a kit utilizing the detection method and application of the kit in the immunological detection of the recurrent spontaneous abortion.

The invention relates to an indirect method for using a monoclonal antibody SPA red cell rosette to detect lymphocyte subsets. The method firstly prepares a primary antibody sensitized red blood cell preservation solution and a secondary antibody sensitized red blood cell preservation solution; secondly, the peripheral blood lymphocytes are separated, and then a secondary antibody sensitized red blood cell application solution and a primary antibody sensitized lymphocyte suspension are prepared by a calcium-free and magnesium-free Hanks balanced salt solution which contains 20 percent newborn calf serum; then the primary antibody sensitized lymphocyte suspension and the secondary antibody sensitized red blood cell application solution are mixed by the volume ratio of 1: 1 to form a mixed suspension; a centrifugation is carried out after the mixed suspension is statically settled under 18 - 28 DEG C, and then the mixed suspension is statically settled for 15 - 45 minutes at the temperature of 4 DEG C; finally, a pipette which is provided with a suction head is used for blowing and sucking the mixed suspension for 8 - 15 times, then cell smears are prepared and calculated by a high-power lens after natural drying and dyeing. Compared with the original McAb-A-E indirect method, the invention is characterized by simplified inspection process, short period, quantized control and satisfying the requirements of clinical inspection.

A method to characterize a T-cell response of a final population of T-lymphocytes resulting from the co-incubation of an initial population of T lymphocytes with a composition of antigen-presenting cells (APCs), said method comprising the steps of a) simultaneous measuring on a single cell basis at least two parameters: (i) proliferation of T lymphocytes and (ii) presence of a T cell antigen receptor on the surface of T lymphocytes and / or presence of at least one biological molecule produced by T lymphocytes, and the attribution of a positive or a negative value to each of these parameters, and b) classification of the final T-lymphocytes population into 2n different subsets of T lymphocytes, n being the number of parameters, each subset being characterized by a positive or a negative value respectively to each parameter and the determination of the proportion of T lymphocytes present in each subset with respect to the number of T lymphocytes in the final population, with said proportion being characteristic of the T-cell response. Use of the method for batch release assay and potency assay.

The invention discloses a construction method and application of a stem cell quantitative standard for screening immunomodulatory ability of high-quality human umbilical cord mesenchymal stem cells. The construction method comprises the following steps: step 1, carrying out primary isolation and amplification on umbilical cord mesenchymal stem cells from a plurality of human umbilical cords, and subculturing to P5 generation; step 2, carrying out flow analysis on the Th1 subgroup and the Th17 subgroup of the lymphocyte subgroup and carrying out flow analysis on the regulatory T cell subgroup of the lymphocyte subgroup; 3, step analyzing a streaming result obtained in the step 2 by adopting Flowjo software, and calculating the proliferation inhibition ratio of Th1, the proliferation inhibition ratio of Th17 and the proliferation promotion ratio of Treg of each group; and respectively averaging to obtain a corresponding stem cell quantitative standard for screening the immunomodulatory ability of the high-quality human umbilical cord mesenchymal stem cells. The invention provides a cell selection standard for basic research and clinical transformation of stem cells for treating autoimmune diseases and immune disorder related diseases, and improves the treatment effect.

The present invention discloses soluble isoforms of CD5 and CD6, which are scavenger-like lymphocyte receptors, for use in the prophylaxis or therapy of disorders or in therapeutic interventions, which would benefit from an exacerbated immune response to either endogenous or exogenous antigens, resulting from a decrease in lymphocyte subpopulations with regulatory functions and / or increase in lymphocyte subpopulations with effector functions. Special disorders are cell growth disorders, and chronic infections of bacterial, viral, fungal or parasitic origin. The invention also provides animal models for the study and the prophylaxis / treatment of autoimmune diseases, cancer, and chronic infections.

The invention discloses a method for effectively removing mononuclear cell interference in lymphocyte subpopulation automatic analysis. The method comprises the following steps of determining fluorescence intensity of CD3 through CD3, SSC-A, CD45, a density-based clustering algorithm and a nuclear density estimation algorithm, identifying T lymphocytes, determining two-dimensional distribution areas of the lymphocytes on the CD45 and the SSC-A and corresponding position parameters, respectively fitting density curves of cells distributed on CD45 and SS in combination with a nuclear density estimation algorithm, determining density curve thresholds of monocytes and lymphocytes distributed on SS and quantiles of CD45 distribution of T lymphocytes, fitting distribution of lymphocyte T cells on CD4 by utilizing nuclear density estimation, and determining CD4 non-expression and weak expression thresholds; and on the basis of determining the thresholds, performing cell grouping on the multi-dimensional data of SSC-A, CD4, CD45, CD16 and CD19, and distinguishing non-T lymphocytes and monocytes according to the thresholds of SSC-A, CD45 and CD4. According to the method, noise interference can be effectively removed.

The invention relates to a system and method for researching an immunosenescence mechanism of an HIV infected person. The system comprises a sample collection unit which is used for obtaining a wholeblood sample and plasma sample of the HIV infected person, a first detection unit which is used for detecting the content of a HIV-1DNA virus library in the whole blood sample, a second detection unitwhich is used for detecting the content of T lymphocyte subsets related to immunosenescence, a third detection unit which is used for detecting the content of cytokines related to body inflammation and homeostasis in the plasma sample, and an analysis unit which is used for receiving and analyzing the correlation and mutual influence degree among the HIV-1DNA, the T lymphocyte subsets and the cytokines. The detection system and method covering nucleic acid, cells and cytokines are established, and the mechanism and process of immunosenescence of the HIV infected person are reflected more completely from the whole in combination with statistical analysis, so that the objective scientific support is provided for the research and development of HIV medicines and the evaluation of medicationeffects or effects of other intervention means, and the reconstruction of the immune function of the HIV infected person is also facilitated.

The invention relates to an application of ursolic acid to antitumor immunity. The ursolic acid has the functions of inhibiting the proliferation and inducing the apoptosis of the hepatoma carcinoma cell of a mouse. The in vivo experiments on the mouse show that the ursolic acid has the function of effectively inhibiting the proliferation of the transplanted hepatocellular carcinoma and has no significant side and toxic effects on the mouse. The ursolic acid has the functions of inhibiting the spleen index which increases abornormally due to hepatic tumor, promoting the proliferation of the T lymphocyte and the B lymphocyte significantly, increasing the proportion of the content of the CD4<+>T lymphocyte subset to that of the content of the CD4<+> / CD8<+T> lymphocyte subset, promoting the expression of IL-2 (interleukin-2) which is a serum cell factor, the TNF-alpha (tumor necrosis factor alpha) and reducing the expression of the IL-4 (interleukin-4). The ursolic acid is similar to the cyclophosphamide which is a low-dose antitumor medicament in the antitumor effect and the immunoregulation function, has low toxicity and can be applied to antitumor immunity to provide a theoretical basis for the development of an antitumor immunity medicament.

The invention discloses a method for inhibiting dimethylbenzanthracene-induced rat mammary cancer with fucoidan, which relates to the technical field of mammary cancer inhibition. Sixty SPF (Specific Pathogen Free ) grade female SD (Sprague-Dawley) rats are randomly divided into a blank control group, a model control group, a low-dose fucoidan intervention group and a high-dose fucoidan intervention group according to body weights, fifteen rats in each group, the rats are killed after sixteen weeks, the incubation period and tumor inhibition rate of the mammary cancer of each group of rats are calculated, and thymus indexes, spleen indexes and the like are calculated. ELISA (enzyme-linked immunosorbent assay) is adopted to assay the levels of interleukins IL-6, IL-10 and IL-12, interferon-Gamma and transforming growth factor-Beta in serum, and a flow cytometer is utilized to assay the activity of NK (natural killer) cells in the peripheral blood of the rats and the cell percentage of the T-lymphocytes subset. The method has the following advantages that the method can regulate the immunity of rats, enhance antitumor immunity and relieve the immunosuppression of tumors, and the regulation of the activity of immune cells by fucoidan is one of the mechanisms of fucoidan in inhibiting the occurrence of mammary cancer.

The invention provides a porcine CD127 polypeptide. The porcine CD127 polypeptide is an extracellular partial fragment of a porcine CD127 protein. In a process of screening a monoclonal antibody, thepolypeptide is used as an immunogen, and in combination with Elisa and a flow cytometer for detection, the monoclonal antibody capable of being used for detecting the porcine CD127 gene through the flow cytometer is prepared. By utilizing the antibody, detection and typing of porcine lymphocyte subpopulations, particularly Treg cells, can be carried out by the flow cytometer while the cell activity is maintained. In order to further research the functions of porcine lymphocytes, research and application of cellular immunity are carried out on pigs, and further research on the relationship between porcine disease resistance characters and lymphocytes has great practical significance.

The invention discloses a method for detecting an immune regulation function of neural stem cells. The method comprises the following steps of (1) conducting adherent culture of suspended neural spheres; (2) inactivating the neural stem cells; (3) marking hPBMC by CFSE; (4) conducting co-culture; (5) conducting lymphocyte subpopulation and proliferation detection; (6) conducting cytokine detection; (7) conducting data processing; and (8) conducting result analysis. According to the method, the hPBMC is utilized to simulate human immune response in vitro, and immunomodulatory effects of the neural stem cells with different sources, different generations, different culture modes and the like are studied, so that a basis is provided for process development of the neural stem cells. Accordingto the method, the nerve spheres subjected to suspension culture are digested into single cells, and then wall attachment is carried out, so that quantity control and inactivation of the single cellsare facilitated, and the proliferation condition of specific cell subsets can be visually detected by using a flow cytometry in combination with different lymphocyte subset antibodies by utilizing thecharacteristic that the CFSE is averagely distributed in a filial generation along with cell division. Various detection indexes are combined, so that operation steps are reduced, experimental materials are saved, and the cost is reduced.

The invention discloses a kit for quantitatively detecting lymphocyte subsets and a detection method thereof. The kit provided by the invention comprises a mixture of six monoclonal antibodies, namelyCD3 FITC, CD56 FITC, CD45 PerCP-Cy5.5, CD8 PE, CD19 PE and CD4 PE-Cyanine7, and a red blood cell lysis buffer. The detection method comprises the following steps: taking a monoclonal antibody mixturein the kit and 100 [mu]L of peripheral blood, incubating at room temperature in a dark place, adding the red blood cell lysis buffer, incubating at room temperature in a dark place, adding a PBS solution, incubating at room temperature in a dark place to obtain a detection sample, detecting the detection sample by an up-flow cytometer, and determining the percentage of each lymphocyte subset. Thekit provided by the invention can be used for quantitatively detecting lymphocyte subsets, can reduce the requirements on the model of a convective cytometer, and is simple to operate and wide in application range.

The present invention discloses soluble isoforms of CD5 and CD6, which are scavenger-like lymphocyte receptors, for use in the prophylaxis or therapy of disorders or in therapeutic interventions, which would benefit from an exacerbated immune response to either endogenous or exogenous antigens, resulting from a decrease in lymphocyte subpopulations with regulatory functions and / or increase in lymphocyte subpopulations with effector functions. Special disorders are cell growth disorders, and chronic infections of bacterial, viral, fungal or parasitic origin. The invention also provides animal models for the study and the prophylaxis / treatment of autoimmune diseases, cancer, and chronic infections.

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a compound traditional Chinese medicine composition for regulating tumor immunity and resisting tumors as well as a preparation method and application thereof. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 5-15 parts of lucid ganoderma, 15-25 parts of astragalus membranaceus, 5-15 parts of ginseng, 5-15 parts of roasted rhizoma atractylodis macrocephalae, 5-15 parts of poria cocos, 5-15 parts of angelica sinensis, 5-15 parts of radix sophorae flavescentis and 5-15 parts of honey-fried licorice root. The compound traditional Chinese medicine composition provided by the invention has the effects of combining purgation and tonification, replenishing qi and enriching blood, detoxifying and removing stasis, regulating tumor immunity, resisting tumors and the like, can regulate and control tumor immune checkpoint proteins, has a killing effect on various tumor cells, can regulate the proportion of a plurality of T lymphocyte subpopulations, causes remarkable increase of immunocompetent cells, and has a remarkable curative effect on tumor cells. The immune function is enhanced, and the effect of treating tumors is achieved.

The invention belongs to the technical field of traditional Chinese medicines, and particularly relates to a novel application of ginseng root-strengthening oral liquid for preventing or treating sicca syndrome. The ginseng root-strengthening oral liquid is an exclusive variety of the South Thick Pharmacy Co., Ltd., and has the effects of nourishing yin and tonifying qi, strengthening root and strengthening primordial qi, clearing heat and cooling blood, promoting blood circulation to remove blood stasis, nourishing yin and generating body fluid, nourishing yin and moistening dryness, tonifying liver and kidney and the like. Tests prove that the traditional Chinese medicine composition can remarkably improve salivary secretion of model rats, improve T lymphocyte subgroups in peripheral blood of mice and reduce the level of immune globulin in serum, and can be used for preparing the medicine for treating or preventing sicca syndrome.

The present invention provides isolated cell compositions for the treatment of cancer, including hematological and solid tumors, comprising a selected, fixed ratio of multiple ex vivo activated lymphocytic cell subsets, including specific immune effector cells directed to specific tumor associated antigens (TAAs), viral associated tumor antigens (VATA), glycolipids, or a combination thereof. By selecting specific fixed ratios of different lymphocytic cell subsets, an immune response which is comprehensive and broad pin biological and immune effector function is provided, enhancing the ability of the administered cells to mount an effective and robust immune response.

The invention discloses an expansion method of various lymphocyte subpopulations and application of the expansion method to preparation of an adoptive immunotherapy medicine for cancers. The expansion method comprises the following steps: S1, adding IFN gamma and zoledronic acid into a culture medium of PBMC collected in the blood of a tumor patient, and re-adding an OKT3 antibody and recombinant human interleukin-2 after culture to stimulate PBMC so as to finish preliminary expansion; S2, after finish of preliminary expansion, mixing immune lymphocytes with feeder layer cells, and after adding zoledronic acid, an OKT3 antibody and recombinant human interleukin-2, continually performing expansion. By adoption of the expansion method, after twice expansion, the sum of the immune cells can be 10,000 to 80,000 times by expansion, the expanded immune cells include alpha beta T cells, gamma delta T cells, NKT cells and NK cells, and tumor cells can be directly killed, or a cellular immunotherapy drug is prepared to kill the tumor cells.

The invention relates to use of oviductus ranae in preparing medicament for curing and preventing diseases caused by immunologic dysfunction. The flow cytometry and the ELISA method proves that the oviductus ranae has a function for immunity adjustment by the results of mouse spleen T-lymphocyte subsets CD3, CD4, CD5, CD8 and the ratio of CD4 / CD8, namely, when the immunity is high, the oviductus ranae has a function for weakening the immunity, and when the immunity is low, the oviductus ranae has a function for enhancing the immunity. The new use of the oviductus ranae proves that the oviductus ranae has a therapeutical effect for the diseases caused by the immunologic dysfunction such as asthma, autoimmune disease, hepatitis and tumor, thereby developing the use of the oviductus ranae inpreparing medicament for curing and preventing the diseases caused by the immunologic dysfunction.

A method for effectively removing mononuclear cell interference in the automatic analysis of lymphocyte subsets, through CD3, SSC‑A, CD45 and a density-based clustering algorithm, plus a kernel density estimation algorithm to determine the strength of the fluorescence intensity of CD3, identify For T lymphocytes, determine the two-dimensional distribution area of ​​lymphocytes on CD45 and SSC‑A and the corresponding position parameters, and combine with the kernel density estimation algorithm to fit the density curves of the distribution of cells on CD45 and SS, respectively, to determine monocytes and lymphocytes The threshold value of the density curve distributed on SS and the quantile of CD45 distribution of T lymphocytes were used to estimate the distribution of lymphoid T cells on CD4 by using kernel density estimation to determine the threshold value of non-expression and weak expression of CD4; on the basis of determining the threshold value, The multi-dimensional data of SSC‑A, CD4, CD45, CD16 and CD19 were classified into cell groups, and non-T lymphocytes and monocytes were distinguished according to the thresholds of SSC‑A, CD45 and CD4. The invention can effectively remove noise interference.

The invention belongs to the field of immunological technique and discloses a method for lymphocyte subsets and a kit thereof. The method of lymphocyte subsets provided by the invention includes T lymphocyte subsets and B lymphocyte subsets. T lymphocyte subsets contain the following steps of: getting different fluorescence labeled antibodies, mixing with a sample to be detected, incubating, detecting by flow cytometry to obtain test data and analyzing the test data. B lymphocyte subsets contain the following steps of: getting different fluorescence labeled antibodies, mixing with a new aforementioned sample to be detected, incubating, detecting by flow cytometry to obtain test data and analyzing the test data. The method provided by the invention is used to realize a more comprehensive fine immunophenotyping of lymphocyte, has advantages of less sample needed to be detected, simple operation, short time needed and high precision, and can be widely applied in immunophenotyping of lymphocyte subpopulation.

The present invention relates to a method, in particular an in vitro method, for identifying CD8 positive subpopulations of a mammal, wherein said method comprises analyzing the bisulfite convertibility of at least one CpG position in the CD8 beta and CD8 alpha cell specific bisulfite convertibility gene region according to SEQ ID No. 1 and 2, wherein a bisulfite convertibility of at least one CpG position in said gene regions is indicative for a CD3+CD8+ and / or CD3+ / −CD8+ cell. The analyzes according to the invention can identify CD3+CD8+ and / or CD3+ / −CD8+ cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood cells.

The invention relates to a composition for regulating body immunologic balance of cancerous persons. The composition is characterized by comprising an immunologic regulation component, an inflammationregulation component, a micro-environment regulation component and an intestinal regulation component; the preparation method includes sequentially covering the intestinal regulation component, the immunologic regulation component (which refers to BCG-PSN preserved in an injector independently), the micro-environment regulation component and the inflammation regulation component (which refers tocamptothecin and anti-tumor RNA respectively preserved in different injectors) sequentially from inside to outside with capsules to separate the four components mutually; the application is to take the composition on an empty stomach. As BCG-PSN, camptothecin and anti-tumor RNA are administrated by injecting, regulation results can be balanced integrally and immunity index of lymphocyte subpopulation can return to normal.

The invention relates to an indirect method for using a monoclonal antibody SPA red cell rosette to detect lymphocyte subsets. The method firstly prepares a primary antibody sensitized red blood cell preservation solution and a secondary antibody sensitized red blood cell preservation solution; secondly, the peripheral blood lymphocytes are separated, and then a secondary antibody sensitized red blood cell application solution and a primary antibody sensitized lymphocyte suspension are prepared by a calcium-free and magnesium-free Hanks balanced salt solution which contains 20 percent newborncalf serum; then the primary antibody sensitized lymphocyte suspension and the secondary antibody sensitized red blood cell application solution are mixed by the volume ratio of 1: 1 to form a mixed suspension; a centrifugation is carried out after the mixed suspension is statically settled under 18 - 28 DEG C, and then the mixed suspension is statically settled for 15 - 45 minutes at the temperature of 4 DEG C; finally, a pipette which is provided with a suction head is used for blowing and sucking the mixed suspension for 8 - 15 times, then cell smears are prepared and calculated by a high-power lens after natural drying and dyeing. Compared with the original McAb-A-E indirect method, the invention is characterized by simplified inspection process, short period, quantized control and satisfying the requirements of clinical inspection.