苯并噻唑衍生物BD960抗T细胞增殖的机制研究

发布时间：2018-12-10 14:56

【摘要】：目的细胞高通量筛选发现苯并噻唑衍生物BD960具有免疫抑制活性,文中探讨BD960抑制T细胞增殖的作用机制。方法免疫磁珠纯化人外周血T细胞,Anti-CD3/CD28m Abs或同种异型抗原活化T细胞。流式细胞术检测BD960对活化T细胞的增殖抑制作用、静息T细胞的细胞毒性作用、CD25表达及细胞周期。酶联免疫吸附法测定BD960对活化T细胞分泌IL-2、IL-4、IL-6、IL-10、IL-17A及IFN-γ等细胞因子的影响。结果 BD960抑制Anti-CD3/CD28 m Abs或同种异型抗原刺激的T细胞增殖,IC50分别为(2.3±0.3)μmol/L和(2.5±0.3)μmol/L,且浓度达100μmol/L也不影响静息T细胞和PBMC的存活。Anti-CD3/CD28 m Abs刺激T细胞72 h表达的CD25比为69.7%,BD960在(0.625、2.5、10)μmol/L均不抑制活化T细胞表达CD25,而0.1μmol/L FK506可抑制CD25表达低至9.4%。活化96 h的T细胞,G0/G1期细胞比为58.5%。BD960能阻滞细胞周期于G0/G1期,并随着浓度的提高G0/G1比例增大。BD960在(0.625、2.5、10)μmol/L能抑制活化T细胞分泌IFN-γ、IL-6和IL-17并呈剂量依赖效应,但不影响IL-2、IL-4和IL-10。结论 BD960抑制作用在T细胞增殖为细胞活化的后期,同时抑制IL-6、IL-17A和IFN-γ等促炎细胞因子的分泌,抑制作用不同于FK506。BD960有望作为先导化合物开发新型免疫抑制剂。
[Abstract]:Objective to identify the immunosuppressive activity of benzothiazole derivative BD960 by high throughput screening. The mechanism of BD960 inhibiting T cell proliferation was discussed in this paper. Methods T cells were purified from human peripheral blood by immunomagnetic beads and activated by Anti-CD3/CD28m Abs or alloantigen. The effects of BD960 on the proliferation of activated T cells, the cytotoxicity of resting T cells, the expression of CD25 and the cell cycle were detected by flow cytometry. Enzyme linked immunosorbent assay (Elisa) was used to determine the effect of BD960 on the secretion of cytokines such as IL-2,IL-4,IL-6,IL-10,IL-17A and IFN- 纬 by activated T cells. Results BD960 inhibited the proliferation of T cells stimulated by Anti-CD3/CD28 m Abs or alloantigen, IC50 was (2.3 卤0.3) 渭 mol/L and (2.5 卤0.3) 渭 mol/L, respectively. The concentration of 100 渭 mol/L did not affect the survival of resting T cells and PBMC. The CD25 ratio of Anti-CD3/CD28 m Abs stimulated T cells for 72 h was 69.7% and BD960 did not inhibit the expression of CD25, by activated T cells at (0.625 渭 mol/L) 渭 mol/L. 0. 1 渭 mol/L FK506 could inhibit the expression of CD25 as low as 9. 4%. When T cells were activated for 96 h, 58.5%.BD960 could block the cell cycle in G0/G1 phase. The ratio of G0/G1 increased with increasing concentration. BD960 at (0.625 渭 mol/L) 渭 mol/L inhibited the release of IFN- 纬, IL-6 and IL-17 from activated T cells in a dose-dependent manner, but did not affect IL-2,IL-4 and IL-10.. Conclusion BD960 inhibits T cell proliferation at the late stage of cell activation and inhibits the secretion of proinflammatory cytokines such as IL-6,IL-17A and IFN- 纬. The inhibitory effect is different from that of FK506.BD960 in the development of novel immunosuppressants.
【作者单位】：成都医学院检验医学院;成都医学院临床医学院;成都医学院科研实验中心;
【基金】：国家自然科学基金(81273530;81302786;81402944) 四川省高校科研创新团队(13TD0028) 四川省教育厅科研项目(15ZB0233;15ZB0235)
【分类号】：R96

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