Objective To investigate the suppression effect and mechanism of Astilbin on lung allograft rejection in rats, in order to know the function of Astilbin on rats’ lung acute rejection. Methods The model of rat left lung transplantation was set up. Sixty lung transplanted rats were divided into two groups randomly, control group: rats were fed with normal saline 1ml per day, experimental group: rats were fed with Astilbin 1mg/kg per day. Survival time, transforming rate of T cells in spleen, activity of interleukin 2 (IL-2) in spleen lymph cells and apoptosis of T cells were observed. Changes in ultrastructure of pulmonary arteries were observed by electron microscope. Results The survival time in experimental group was prolonged than that in control group (25.4±2.1 d vs. 13.4±1.2 d;t=2.042, Plt;0.05）. Transforming rate of T cells of spleen in experimental group was significant lower than that in control group (23 465.8±8 783.4 cpm vs. 74 567.3±12 874.6 cpm; t=2.284，Plt;0.05）.Activity of IL-2 of spleen lymph cells in experimental group was significant lower than that in control group (425±2.65U/ml vs. 23.46±1.82U/ml; t=3.165, Plt;0.01).Effectively derive apoptosis of activated T cells in acute rejection were observed in experimental group, the ultrastructure of pulmonary arteries showed attenuated injury in experimental group. Conclusion Astilbin decreased the IL-2 concentration in plasma and induced the apoptosis in activated T cells, then suppressed the acute rejection of lung allograft and prolonged the survival period of lung transplantation rats.
Autoimmune ocular diseases are a type of inflammatory eye condition characterized by the involvement of the immune response. This includes various types disease such as autoimmune uveitis, thyroid-associated eye disease, and primary Sj?gren's syndrome. In recent years, breakthroughs have been achieved in inducing transplant tolerance, understanding tumor immune evasion, and preventing autoimmune diseases using immune checkpoint molecules. Negative immune checkpoints effectively control disease progression by inhibiting T cell proliferation, reducing inflammatory cytokine levels, and ultimately regulating autoimmune balance. Therefore, the negative immune checkpoint molecules are expected to be used as a new therapeutic target in the future, and the combination therapy through the combination of negative immune checkpoint drugs is expected to become an important direction to improve the efficacy of the treatment of autoimmune diseases.
【 Abstract】 Objective To construct a lentiviral expression vector carrying Nogo extra cellular peptide residues 1-40(NEP1-40) and to obtain NEP1-40 efficient and stable expression in mammalian cells. Methods The DNA fragment ofNEP1-40 coding sequence was ampl ified by PCR with designed primer from the cDNA l ibrary including NEP1-40 gene, and then subcloned into pGC-FU vector with in-fusion technique to generate the lentiviral expression vector, pGC-FU-NEP1-40. The positive clones were screened by PCR and the correct NEP1-40 was confirmed by sequencing. Recombinant lentiviruses were produced in 293T cells after the cotransfection of pGC-FU-NEP1-40， and packaging plasmids of pHelper 1.0 and pHelper 2.0. Green fluorescent protein (GFP) expression of infected 293T cells was observed to evaluate gene del ivery efficiency. NEP1-40 protein expression in 293T cells was detected by Western blot. Results The lentiviral expression vector carrying NEP1-40 was successfully constructed by GFP observation， and NEP1-40 protein expression was detected in 293T cells by Western blot. Conclusion The recombinant lentivirus pGC-FU-NEP1-40 is successfully constructed and it lays a foundation for further molecular function study of NEP1-40.
Objective
To summarize research progress of the mechanism of natural killer cells (NK cells) acted in regulating the T cell immunity in chronic infectious disease.
Method
Literatures about recent studies concerning how NK cells act as a regulator for T cells in chronic infectious disease were reviewed according to the results obtained from PubMed, Embase, CNKI, CBM, and Wanfang databases.
Results
NK cells that acted as regulators of T cell immunity could affect T cell immune responses through influencing antigen presentation, secreting cytokine, and presenting lytic activities, thus playing an important role in the immunological therapy of chronic infectious diseases.
Conclusion
NK cells are critical for T cell immune regulation, which could provide noval strategies for immunological therapy of chronic infectious disease, transplantation-related immune rejection, and autoimmune disease.
Objective To investigate the clinical implication on expression of HLA class I in breast cancer tissures.Methods The expression of HLA class I in 271 patients with breast cancer that underwent radical operation was examinedby using immunohistochemically， and the correlation between the expression of HLA class I and clinicalpathological characteristics and prognosis of breast cancer was analyzed. Results The b positive expression of HLA class I in breast cancer tissures was observed in 92 patients （33.9%）， the expressions of HLA class I in 179 patients （66.1%）were downregulation. The expression of HLA class I expression in breast cancer tissures was significantly associated with the axillary lymph node metastasis， TNM stage （P＜0.05）， other lymph node metastasis， and vascular invasion （P＜0.05）. The disease free survival rate of patients with positive expression of HLA class I was higher than that expression downregulation of HLA class I （P＜0.05）. Conclusion The examination of HLA class I expression is useful for the prediction of tumor progression and recurrent risk of breast cancer via the antitumor immune system.
【Abstract】ObjectiveTo find a stable and efficient method for ex vivo concentration of CD4+CD25+ regulatory T cells in rats. MethodsCD4+CD25+ regulatory T cells were separated from the rat splenic cells in two steps by magnetic cell sorting (MACS) system. CD4+ T cells were first negatively sorted by cocktail antibodies and antiIgG magnetic microbeads. And then CD4+CD 25+T cells were positively sorted by antiCD25 PE and antiPE magnetic microbeads. The purity and the cell survival rate of the sorted cells were measured by flow cytometry and trypan blue dyeing respectively, and the immunosuppressive action of CD4+CD25+ T cells on the proliferation of CD4+CD25- T cells was also assessed by in vitro cell proliferation assay. ResultsThe purity of negatively sorted CD4+ T cells were (83.6±2.5)% (79%~87%), and the purity of positively sorted CD4+CD25+ T cells was (90.2±1.8)% (86%~93%) with the survival rate of (92.8±3.4)% (92%~95%). These concentrated cells significantly suppressed the proliferation of CD4+CD25- T cells in mixed lymphocyte culture （CD25+/CD25- versus CD25-, P＜0.01）. ConclusionWe created a twostep procedure of magnetic cell sorting system for CD4+CD25+ regulatory T cells sorting, which insures the cells to be satisfactorily purified and well functioned.
Objective To investigate the role of T helper 17 ( Th17) cells and CD4 + CD25 + Foxp3+regulatory T cells ( Treg) in the pathogenesis of asthma in a mouse model. Methods Twenty-four BALB/ c mice were randomly divided into an asthma group and a normal control group, with 12 mice in each group.Asthma model was established by ovalbumin sensitization and aerosol challenge in the asthma group. Airway reactivity was measured by plethysmography. The total and differential cell counts in bronchoalveolar lavage fluid ( BALF) were measured. The ratio of Th17 and Treg cells to mononuclear cells in the spleens of mice were detected by flow cytometry. The levels of IL-17 and IL-10 in BALF and lung homogenates were measured by ELISA. Results The bronchial provocation test showed that the average lung resistance increased remarkably in the asthma group. In spleens of the asthmatic mice, the percentage of Th17 cells was significantly higher [ ( 5.68 ±1. 99)% vs ( 2.80 ±0. 82) %, P lt; 0. 01] , and the percentage of Treg cells was significantly lower [ (2.88 ±0. 46) % vs ( 6.10 ±2.44) % , Plt; 0. 01] , with the ratio of Th17 to Treg significantly increased( 1. 93 ±0. 41 vs 0. 50 ±0. 15,P lt;0. 01) . In BALF and lung homogenates of the asthma group, the level of IL-17 was significantly higher[ ( 22. 37 ±3. 00) pg/mL vs ( 11. 42 ±2. 15) pg/mL, ( 52. 93 ±5. 39) pg/mL vs ( 19. 38 ±2. 65) pg/mL, both Plt; 0. 01] , and the level of IL-10 was significantly lower[ ( 6. 05 ±1. 25) pg/mL vs ( 14. 23 ±2. 94) pg/mL, ( 9. 33 ±1. 79) pg/mL vs ( 21. 40 ±2. 44) pg/mL, both P lt; 0. 01] compared with the control group.Conclusion The imbalance of Th17/ Treg plays an important role in the pathogenesis of asthma.
ObjectiveTo investigate the diagnostic value of tuberculosis T cell spot test (T-SPOT.TB) and erythrocyte sedimentation rate (ESR) test in the diagnosis of simple pulmonary nodules in Xinjiang.MethodsA retrospective analysis of 72 patients with asymptomatic simple pulmonary nodules in the Department of Thoracic Surgery, the First Affiliated Hospital of Xinjiang Medical University from October 2017 to July 2019 was performed. According to the pathological results, the patients were divided into a tuberculoma group [n=23, including 14 males and 9 females, aged 37-84 (56.91±12.73) years] and a lung cancer group [n=49, including 31 males and 18 females, aged 34-83 (61.71±10.15) years]. The predictive value of T-SPOT.TB and ESR results for the diagnosis of simple pulmonary nodules was evaluated.ResultsThe positive rate of T-SPOT.TB in the tuberculoma group (69.60%) was higher than that in the lung cancer group (42.90%) (χ2=5.324, P=0.021), with a sensitivity of 69.56% and specificity of 57.14%; the positive ESR was 47.80% in the tuberculoma group and 59.20% in the lung cancer group, and no statistical difference was found between the two groups (χ2=0.981, P=0.322), with a sensitivity of 47.82% and specificity of 40.81%; the area under receiver operating characteristic curve (AUC) was 0.618, the 95% confidence interval of AUC was (0.479, 0.758), and the Youden’s index was 0.267 with a sensitivity of 69.60% and specificity of 57.10%. Difference in the T-SPOT.TB and ESR test results was statistically significant (χ2=4.986, P=0.026), but the correlation between the tests was weak with a Pearson contingency coefficient of 0.199. ESR results in patients with different ages were statistically different (χ2=7.343, P=0.025), but the correlation between age and ESR results was weak with a Pearson contingency coefficient of 0.239; T-SPOT.TB results in patients with different ages were not statistically different (χ2=0.865, P=0.649), and the correlation between age and ESR results was weak with a Pearson contingency coefficient of 0.084.ConclusionThe diagnostic value of T-SPOT.TB and ESR tests is small in the diagnosis of simple pulmonary nodules.
The specific binding of T cell receptors (TCRs) to antigenic peptides plays a key role in the regulation and mediation of the immune process and provides an essential basis for the development of tumour vaccines. In recent years, studies have mainly focused on TCR prediction of major histocompatibility complex (MHC) class I antigens, but TCR prediction of MHC class II antigens has not been sufficiently investigated and there is still much room for improvement. In this study, the combination of MHC class II antigen peptide and TCR prediction was investigated using the ProtT5 grand model to explore its feature extraction capability. In addition, the model was fine-tuned to retain the underlying features of the model, and a feed-forward neural network structure was constructed for fusion to achieve the prediction model. The experimental results showed that the method proposed in this study performed better than the traditional methods, with a prediction accuracy of 0.96 and an AUC of 0.93, which verifies the effectiveness of the model proposed in this paper.
【Abstract】Objective To explore the feasibility that the recipient against donor antigen-specific T lymphocytes clones are formed,and the suicide genes are induced into the clone. In the end it may induce the transplanted-organ tolerance. Methods The recipient rats were immunized by the donor rats-splenocytes, then the recipient’s T cell were isolated, purified and diluted with limited-dilute methods into single cell.The T cells were cultured by adding raise cells,ConA or IL2 under the different concentration and at last the T cell clone were formed. Results A stable recipient against donor antigen-specific T cell clone was established. The difference among the different immune groups was very significant(tgt;t0.05). The T cell clone was not formed without raise cells or only with raise cells. The rare clone could be formed by ConA stimulation but without IL-2. The clone-forming rate was associated with concentration of IL-2. ConclusionThe mature T cell is able to proliferate to form clone when the condition is suitable. The recipient is immunized with donor rats spleen cellular antigen. The T cells clones selected in the end are donor antigen-specific.
