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目的:探讨CD3/CD46共刺激通路对同种移植免疫反应的调控作用及其机制。方法:分离转人CD46的C57BL/6小鼠的脾脏淋巴细胞,采用MACS免疫磁珠分选CD4+T细胞,以阴性对照组作为对照,体外检测CD3、ConA、CD3/CD28、CD3/CD46、CD3/CD28/CD46共刺激对CD4+T细胞增殖的作用效果,并检测其细胞上清液中白细胞介素2(IL-2),γ-干扰素(γ-IFN),白细胞介素10(IL-10)和转化生长因子β(TGFβ-)的水平,并以BALB/c小鼠的脾细胞作为刺激细胞,以转人CD46的C57BL/6小鼠的淋巴细胞作为反应细胞进行同种混合淋巴细胞培养,MMT法检测淋巴细胞增殖活性。结果:与CD3组或阴性对照组相比,ConA、CD3/CD28、CD3/CD46和CD3/CD28/CD46共刺激后,CD4+T细胞均出现显著增殖反应(P<0.05),而且CD3/CD28/CD46共刺激组的增殖活性较CD3/CD28或CD3/CD46共刺激组显著增高(P<0.05)。CD3/CD28共刺激后,IL-2和γ-IFN水平较阴性对照组和CD3组显著升高(P<0.05),CD3/CD46共刺激后,IL-10和TGFβ-水平较阴性对照组、ConA组、CD3组和CD3/CD28共刺激组显著升高(P<0.05)。CD3/CD46共刺激后可以显著地抑制同种混合淋巴细胞的增殖反应(P<0.05)。结论:CD3/CD46共刺激通路可以诱导CD4+T细胞产生IL-10和TGFβ-,抑制同种移植免疫反应的发生。
Objective: To investigate the regulatory effect of CD3 / CD46 costimulatory pathway on allograft immune response and its mechanism. Methods: The splenic lymphocytes of C57BL / 6 mice which were transplanted to CD46 were isolated and the CD4 + T cells were sorted by MACS magnetic beads. The negative control group was used as a control. CD3, ConA, CD3 / CD28, CD3 / CD3 / CD28 / CD46 costimulation on the proliferation of CD4 + T cells. The levels of interleukin 2 (IL-2), interferon-γ (IFN- γ) and interleukin 10 IL-10) and transforming growth factor β (TGFβ-) were measured. The splenocytes of BALB / c mice were stimulated with lymphocytes of CD46-transfected C57BL / 6 mice as the same cells Lymphocyte culture, MMT assay lymphocyte proliferation activity. Results: Compared with CD3 group or negative control group, CD4 + T cells showed significant proliferative responses (P <0.05) after ConA, CD3 / CD28, CD3 / CD46 and CD3 / CD28 / / CD46 costimulation group was significantly higher than that of CD3 / CD28 or CD3 / CD46 costimulation group (P <0.05). Compared with negative control group and CD3 group, the levels of IL-2 and γ-IFN were significantly increased after CD3 / CD46 costimulation (P <0.05). Compared with negative control group, ConA group, CD3 group and CD3 / CD28 costimulation group were significantly increased (P <0.05). CD3 / CD46 costimulation could significantly inhibit the proliferation of allogeneic mixed lymphocytes (P <0.05). Conclusion: The CD3 / CD46 costimulatory pathway can induce the production of IL-10 and TGFβ- in CD4 + T cells and inhibit the allograft immune response.