36 results about "T-Lymphocyte Subsets" patented technology

A method for the treatment or prevention of autoimmune diseases, allergic or atopic disorders, and graft rejection is provided, comprising inducing the death by apoptosis of a subpopulation of T lymphocytes that is capable of causing such diseases, while leaving substantially unaffected the majority of other T lymphocytes. Cell death is achieved by cycle(s) comprising challenging via immunization these T cells with antigenic substance at short time intervals, or by immunization followed by administering interleukin-2 (IL-2) when these T cells are expressing high levels of IL-2 receptor so as to cause these T cells to undergo apoptosis upon re-immunization with the antigenic peptide or protein. These methods are applicable to the treatment of autoimmune diseases such as, for example, multiple sclerosis, uveitis, arthritis, Type I insulin-dependent diabetes, Hashimoto's thyroiditis, Grave's thyroiditis, autoimmune myocarditis, etc., allergic disorders such as hay fever, extrinsic asthma, or insect bite and sting allergies, food and drug allergies, as well as for the treatment or prevention of graft rejection.

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 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 discloses a method for assaying T-lymphocyte subpopulation of swine peripheral blood by flow cytometry. The method is characterized by including the following steps: swine peripheral venous blood is extracted to prepare anticoagulation blood; the anticoagulation blood is sucked, anti-swine CD3, CD4 and CD8 monoclonal antibodies are respectively added, vortex mixing is carried out, and dyeing is carried out in the dark at 4 DEG C; a red blood cell lysis buffer is used for rupturing red blood cells; a PBS solution is used for washing and resuspending cells, and a flow cytometer is used for assaying; and an assay result is analyzed, so that a proportion of the T-lymphocyte subpopulations of the swine peripheral blood is obtained. the stable and standard method for assaying the T-lymphocyte subpopulations of the swine peripheral blood by flow cytometry is successfully established; by improving for the problems of target cell choice, compensative regulation and parameter selection in the process of pretreating, assaying and analyzing a sample, errors caused by human factors in the process of an experiment are prevented, and thereby the authenticity and objectivity of result judgment are increased.

The invention discloses a method for detecting avian peripheral blood T lymphocyte subsets by flow cytometry; the method comprises the steps of collecting avian peripheral venous blood, preparing anticoagulant blood, and treating with lymphocyte separate to obtain peripheral blood leukocytes; centrifugally washing the peripheral blood leukocytes with PBS (phosphate buffer solution) to prepare single-cell suspension; sucking the single-cell suspension, adding anti-chicken CD3, CD4 and CD8 monoclonal antibodies, mixing well by cyclone, and staining in the shade at 4 DEG C; after staining, washing with PBS, resuspending cells, and detecting with a flow cytometer; analyzing detection results, the ratio of avian peripheral blood T lymphocyte subsets. The method according to the invention which detects avian peripheral blood T lymphocyte subsets stably and regularly is established, thereby solving the problem that differences between sample pretreatment, adjustment of voltage and fluorescence compensation upon detection and processing modes for result analysis cause high fluctuations of detection results and instability.

The invention discloses a flow cytometry detection method for mouse thymus or spleen T-lymphocyte subsets. The method comprises the following steps: acquiring thymus or spleen tissues of tested mice, and preparing single-cell suspension; respectively sucking the single-cell suspension of the thymus or spleen, respectively adding anti-mouse DC3, CD4 and CD8 monoclonal antibodies, swirling and uniformly mixing, keeping away from light at the temperature of 4 DEG C and staining; washing with PBS (Phosphate Buffer Solution) after staining, resuspending the cells, and detecting by using flow cytometry; and analyzing the detection result, thereby obtaining the percentage of each T-lymphocyte subset of the thymus or spleen of the mice. The stable and standardized method for detecting the mouse thymus or spleen T-lymphocyte subsets through flow cytometry is successfully established in the invention. By improving the problems such as target cell selection, compensative regulation and parameter selection in the sample pretreatment, detection and analysis process, errors caused by human factors in the test process are avoided, and the authenticity and objectivity of result judgment are improved.

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 belongs to the technical field of traditional Chinese medicines, provides a Chinese herba preparation for treating nephrotic syndrome proteinuria and a preparation method thereof, and aims to effectively control proteinuria, delay the evolution of nephrotic syndrome, and effectively regulate imbalance of nephrotic syndrome T lymphocyte subpopulation. The Chinese herba preparation is prepared from astragalus membranaceus, flatstem milkvetch seed, dogwood, Chinese yam, gorgon fruit, angelica sinensis, and leonurus according to certain proportion. According to the Chinese herba preparation, astragalus membranaceus which has functions of invigorating the spleen, supplementing qi, tonifying deficiency and inducing diuresis to alleviate edema is taken as the main raw material, flatstem milkvetch seed and dogwood which have functions of tonifying the liver and kidney and securing essence and reducing urination and the Chinese yam and gorgon fruit which have functions of tonifying the kidney, arresting seminal emission, tonifying the spleen and clearing damp are taken as ministerial drugs, the angelica sinensis and leonurus which have functions of enriching and activating blood and inducing diuresis to reduce edema are taken as auxiliary drugs, and all the components are compatible, so that the Chinese herba preparation can treat both syndromes and root causes, tonify qi, the kidney and the spleen, and activate blood. The experiment research on an adriamycin nephropathy rat model proves that the Chinese herba preparation has obvious protective effect for renal pathological lesion caused by doxorubicin, can be used for treating nephrotic syndrome and is applicable to people with abnormal nephrotic proteinuria immunity.

The invention discloses a method for preparing anti-influenza A virus and novel universal epitope vaccine. The method comprises the following steps: combining a network server and related software to predict functional epitopes of NP, M1 and HA that are related with influenza protection antigen via a bioinformatics method, obtaining a total of 8 epitopes from H1N1, H3N1, H3N2, H5N1, H5N2 subtype CTL epitope and a B-cell epitope, and adding a flexible segment GPGPG between every two of the epitopes to act as linker and naming the linked product as HMN; linking a nucleotide sequence that corresponds to the epitope polypeptide to a plasmid carrier, performing prokaryotic expression, and adding an adjuvant to the purified expressed protein to prepare the vaccine, wherein the vaccine is used to perform immunization in mice. Western blot test proves that the recombinant polypeptide is effective in gene expression and has antigenicity. The T-lymphocyte subset index of the test group is higher than that of the control group in the mice immunization test, and the result indicates that the polypeptide can induce BALC / c mice to generate specific humoral and cellular immune responses specific to the selected epitopes and proves that the polypeptide has strong immunogenicity.

The invention provides an application of flavonoids compounds in the preparation of a T lymphocyte subsets regulating drug. The innovative discovery of the invention is that the flavonoids compounds can regulate the proportion of T lymphocyte subsets, so that the effect of immune system is regulated, and the related pathological injury can be cured. Specifically, the proportion of a CD4+IFN-gamma+Th1 cell subset, a CD4+IL-17+Th17 cell subset and a CD4+CD3e+CD62L+ThO cell subset can be lowered, and the proportion of CD4+CD25+Fo8*p3+Treg cell subset is increased. Chrysin can maintain the integrity of the blood-retinal barrier of an EAU mouse, inhibit the macrophage infiltration, and inhibit the intraocular inflammation. At the same time, the expression quantity of retinas STAT1, STAT3 and phosphorylated protein is inhibited. Based on new properties of the flavonoids, the flavonoids can be applied for curing a plurality of diseases with the effect of proportional imbalance of T cell subsets.

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 assaying T lymphocytes subsets of chicken thymuses by flow cytometry, which includes the following steps: after a chicken is killed, thymus tissues are obtained, and single-cell suspension is prepared; the single-cell suspension is added with 1 dose of anti-chicken CD3 monoclonal antibody, 1 dose of anti-chicken CD4 monoclonal antibody and 1 dose of anti-chicken CD8 monoclonal antibody, vortex mixing is carried out, and staining is carried out under 4 DEG C away from light; stained cell solution is washed with PBS and cells are resuspended, and a flow cytometer is used for assaying; an assay result is analyzed, and thereby a proportion of T lymphocytes subsets of chicken thymuses is obtained. For the issues of target cell choice, compensative regulation and parameter selection in the process of sample processing, assay and analysis, the method for assaying T lymphocytes subsets of chicken thymuses by flow cytometry disclosed by the invention establishes a stable and standard method for assaying T lymphocytes subsets of chicken thymuses.

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 analyses 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 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 discloses a purpose and application of R'-beta-D-fructopyranoside in preparing a medicament for preventing and treating cancer cell diffusion transfer. The medicament for preventing and treating cancer cell diffusion transfer is prepared by firstly utilizing the depression action of R'-beta-D-fructopyranoside on cancer cell diffusion transfer and is clinically applied to preventing and treating cancer cell diffusion transfer of digestive systems such as gastric cancer, liver cancer, intestinal cancer and the like; when matching with concurrent chemoradiotherapy for preventing and treating cancer cell diffusion transfer, the invention achieves the remarkable high-efficiency and low-toxicity effects that the normal retention rate of the hematic convention of a patient is 70 percent, the normal retention rate of blood T lymphocell subgroup is 70 percent, and the normal retention rate of hepatic / renal function is over 80 percent, and solves the problem that no medicaments for preventing and treating cancer cell diffusion transfer appear till now.

The invention discloses a recombinant porcine pseudorabies virus strain expressing a PPV VP2 gene and a porcine IL-18 gene. The recombinant porcine pseudorabies virus (Herpesviridae) strain is numbered as CCTCC NO.V201347. The recombinant porcine pseudorabies virus PGVP218 established by the recombinant porcine pseudorabies virus strain and an IL-18-free recombinant pseudorabies virus rPRV-VP2 established in a laboratory respectively immunize rats, meanwhile a DMEM culture solution is used as a negative control and a PPV HB98 vaccine and a PRV inactivated vaccine are used as a positive control group to immunize the rats, and the immunogenicity of the recombinant virus is studied through PPV specific antibody detection, a PRV antibody neutralization test, peripheral blood T lymphocyte subpopulation determination and a attacking protection test. Results show that the recombinant virus PGVP218 is safe and effective to the rats, can induce the rats to produce specific PPV and PRV antibodies, and the number of CD4+ / CD8+T lymphocyte subpopulations is increased.

The invention provides application of rutin to preparation of a drug for promoting mammal immune efficacy. According to the technical scheme, it is found through experimental means that rutin not only has the pharmacologic effects such as anti-oxidization and lipid metabolism regulation as disclosed in the prior art, but has a specific immune system regulating function. It is found through endosome experiments that after rutin injection, proliferation of T-lymphocytes in animal serum is improved remarkably, the content of CD4+ cells and CD8+ cells and the proportion of CD4+ cells and CD8+ cells in T-lymphocyte subpopulation are increased, the expression quantity of cell factors IFN-gamma and IL-4 is also increased, and therefore the immune system stimulating effect of rutin is verified. On this basis, the immunological enhancement effect, especially immune synergism with vaccines, of rutin for specific diseases is further studied. Experiment results show that by injecting a rutin solution after conducting adenovirus vaccine or foot-and-mouth disease vaccine immunity on animals, antibody titer can be increased remarkably, so that the immune effect of vaccines can be improved.

The invention discloses an experimental method for proving connected relation of estrogen and gap. The method comprises the following steps: measuring animal blood pressure; taking out rat kidney by HE dyeing of kidney tissues; preparing rat peripheral blood lymphocyte suspension; and detecting peripheral blood T lymphocytal subset and Cx0 / Cx43.

The invention provides a homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and an application thereof. The polysaccharide is separated and extracted from a traditional Chinese medicine snakegourd root, is a white flocculent solid, can be easily dissolved in water, and is composed of C, H and O, the specific rotation [alpha]<D><50>(H2O) is +165.58, the weight-average molecular weight (Mw) is 1.76*10<4>, and the main glycosyl group of the polysaccharide is glucose. RTPS-I can promote proliferation of human peripheral blood mononuclear cells (PBMCs), increases T lymphocyte subgroups in the PBMCs, and stimulates the human PBMCs to secrete TNF-alpha and IL-6 cytokines, and the immune activity of the RTPS-I is obviously better than that of snakegourd root total polysaccharides; and the RTPS-I can substantially inhibit the growth of human cervical carcinoma cells (HeLa cells) and human breast cancer cells (MCF-7 cells). The homogeneous snakegourd root polysaccharide can be used for exploiting immunoloregulation adjuvants, and can also be used as an antitumor drug lead compound.

The invention discloses an experiment method for proving a connection relationship between NO and a slit. The method comprises the following steps: tail artery blood pressure monitoring; HE dyeing; and flow cytometry detection of rat peripheral blood T lymphocyte subgroup, and expression of CD4+, CD8+ and up-Cx40. A research result shows that the SHR microvascular endothelia are injured and have inflammatory cell infiltration, accompanied by rat peripheral blood T lymphocyte subgroup disorder and increased expression of the up-Cx40 in T lymphocytes, and NO intervention reduces the injured degree of the microvascular endothelia, corrects the disorder state of the peripheral blood T lymphocyte subgroup and reduces the expression of the up-Cx40 in T lymphocytes, so it is deducted that the NO possibly inhibits the slit between the SHR peripheral blood T lymphocytes to protect cerebral vessels.

The invention discloses a configuration method for obtaining a memorable T lymphocyte subset culture medium. The serum-free medium is formed by adding 40 micrograms / ml of insulin, 10 micrograms / ml oftransferrin, 101 micrograms / ml of ethanol amine, 2 mg / ml of catalase and other cell factors into an RPM1 1640 basic culture medium and using microelements of a Trace element A and a Trace element B cooperatively. The serum-free medium can increase the amplification multiple of T cells and remarkably increase the proportion of memorable T cells to the total T cells, is suitable for in-vitro production of memorable T lymphocyte subset cells, easily improves the efficiency of clearing away tumors in the body of a patient through CAR-T cells, and has positive meanings for CAR-T cell immunotherapy.

The invention provides a homogeneous snakegourd root polysaccharide with immunization activity and antitumor effect, and an application thereof. The polysaccharide is separated and extracted from a traditional Chinese medicine snakegourd root, is a white flocculent solid, can be easily dissolved in water, and is composed of C, H and O, the specific rotation [alpha]<D><50>(H2O) is +165.58, the weight-average molecular weight (Mw) is 1.76*10<4>, and the main glycosyl group of the polysaccharide is glucose. RTPS-I can promote proliferation of human peripheral blood mononuclear cells (PBMCs), increases T lymphocyte subgroups in the PBMCs, and stimulates the human PBMCs to secrete TNF-alpha and IL-6 cytokines, and the immune activity of the RTPS-I is obviously better than that of snakegourd root total polysaccharides; and the RTPS-I can substantially inhibit the growth of human cervical carcinoma cells (HeLa cells) and human breast cancer cells (MCF-7 cells). The homogeneous snakegourd root polysaccharide can be used for exploiting immunoloregulation adjuvants, and can also be used as an antitumor drug lead compound.

The invention discloses a method for preparing anti-influenza A virus and novel universal epitope vaccine. The method comprises the following steps: combining a network server and related software to predict functional epitopes of NP, M1 and HA that are related with influenza protection antigen via a bioinformatics method, obtaining a total of 8 epitopes from H1N1, H3N1, H3N2, H5N1, H5N2 subtype CTL epitope and a B-cell epitope, and adding a flexible segment GPGPG between every two of the epitopes to act as linker and naming the linked product as HMN; linking a nucleotide sequence that corresponds to the epitope polypeptide to a plasmid carrier, performing prokaryotic expression, and adding an adjuvant to the purified expressed protein to prepare the vaccine, wherein the vaccine is used to perform immunization in mice. Western blot test proves that the recombinant polypeptide is effective in gene expression and has antigenicity. The T-lymphocyte subset index of the test group is higher than that of the control group in the mice immunization test, and the result indicates that the polypeptide can induce BALC / c mice to generate specific humoral and cellular immune responses specific to the selected epitopes and proves that the polypeptide has strong immunogenicity.

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.

The invention provides a composition used for relieving ratio imbalance of T lymphocyte subpopulation and a product applying the composition, and relates to the technical field of medicines. A composition of a rhubarb extract and defatted croton seed powder can effectively stop diarrhea. Proved by experiments, the composition of the rhubarb extract and the defatted croton seed powder can relieve the ratio imbalance of the T lymphocyte subpopulation and can activate T cell immune response, so that mucosal barrier injury is protected and / or intervened, and therefore, another evidence is provided for enabling the composition of the rhubarb extract and the defatted croton seed powder to realize mucosal barrier injury repair, and a potential value is achieved for preventing and / or treating ulcerative colitis. The medicine provided by the invention is prepared from the rhubarb extract and the defatted croton seed powder, can be used for relieving the ratio imbalance of the T lymphocyte subpopulation, activating T cell immune response or protecting and / or intervening mucosal barrier injury, and has the advantages of good curative effect and low side effect.

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 invention relates to the field of biological preparations, in particular to a pharmaceutical composition containing rhIL-12 for treating inactive hepatitis B. The pharmaceutical composition, in terms of each milliliter, consists of 10-20 [mu]g of recombinant human interleukin-12, 10-60 [mu]g of a recombinant HBsAg protein vaccine, 5-20 [mu]g of a cow mycobacteria vaccine and the balance of a water solution. The pharmaceutical composition is capable of activating a T achroacyte subgroup, promoting CD3+CD4+ and CD3+CD8+ cells to secrete IFN-[gamma] and inhibiting PD-1 expression of the CD3+CD4+ and CD3+CD8+ cells; in addition, the pharmaceutical composition can also activate mononuclear cells and dendritic cells, promote CD14+ cells and CD1a+ cells to express Toll-like receptors and enhance specific immunity and non-specific immunity functions, so as to break through an immune tolerance state and to better cure HBV infection; and in addition, the pharmaceutical composition can promote HBsAg seronegative conversion.

The invention discloses an experiment method for proving connection relationship between hydrogen sulfide and a gap. The method comprises the following steps: blood pressure measurement of animals, preparation of a rat peripheral blood lymphocyte suspension and detection of peripheral blood T-lymphocyte subsets and Cx40 and Cx43. The relationship between the hydrogen sulfide and protein on T-lymphocyte is disclosed for the first time; the hydrogen sulfide is prompted to communicate through affecting gap junction among immune cells and release of inflammatory factors is adjusted, so that the experiment method plays an important role in improvement of target organ damages caused by hypertensive inflammation.