重组TACI-Ig融合蛋白抑制抗CD3抗体诱导T淋巴细胞增殖与活化

发布时间：2018-01-31 02:02

本文关键词： B淋巴细胞刺激因子重组TACI-Ig融合蛋白 T淋巴细胞免疫治疗自身免疫病　出处：《安徽医科大学学报》2016年07期 　论文类型：期刊论文

【摘要】：目的探讨抗CD3抗体诱导小鼠T淋巴细胞增殖与活化的机制并观察重组人TACI-Ig融合蛋白(rh TACI-Ig)对其的影响。方法免疫磁珠纯化得到小鼠T淋巴细胞,用抗CD3抗体刺激,同时给予rh TACI-Ig、rh TNFR∶Fc或Ig G-Fc。[3H]-TdR参入法检测T细胞增殖能力,流式细胞术检测T细胞亚群比率,Western blot法检测B淋巴细胞刺激因子、BAFF受体(BAFFR)和跨膜激活剂及钙调亲环素配体相互作用分子(TACI)两个受体及IL-2受体(IL-2R)表达水平和NF-κB活性,采用小干扰RNA(siRNA)抑制T细胞BAFFR或TACI的表达。结果抗CD3抗体体外可促进T细胞增殖与活化,BAFF、白细胞介素-2(IL-2)、γ-干扰素(IFN-γ)和转化生长因子-β(TGF-β)分泌,BAFFR、TACI、IL-2R表达升高和NF-κB活性增强(P0.05)。Rh TACI-Ig(0.1、1、10、100μg/ml)体外给药可抑制抗CD3抗体诱导的T淋巴细胞增殖(P0.05);rh TACI-Ig(1、10、100μg/ml)可明显降低抗CD3抗体诱导产生的细胞因子水平(P0.05,P0.01),显著抑制CD4+CD69+T细胞、CD4+CD154+T细胞比率,提高CD4+CD62L+T细胞比率(P0.05,P0.01),且rh TACI-Ig(10、100μg/ml)能降低T细胞上BAFFR、TACI、IL-2受体的表达,抑制NF-κB活性(P0.05)。沉默BAFFR或TACI对抗CD3抗体诱导的T细胞增殖有抑制作用(P0.01)。结论抗CD3抗体部分通过产生BAFF,激活BAFFR信号而促进T细胞增殖与活化,rh TACI-Ig中和BAFF,抑制BAFF相关受体的表达和NF-κB活性,减少T细胞表达IL-2受体,阻止T细胞过度增殖与活化。
[Abstract]:Objective to investigate the mechanism of proliferation and activation of mouse T lymphocytes induced by anti CD3 antibody and to observe the recombinant human TACI-Ig fusion protein rh TACI-IgA. Methods Murine T lymphocytes were purified by immunomagnetic beads. CD3 antibody was used to stimulate rh TACI-IgA rh TNFR:Fc or Ig G-Fc. [3H] -TdR incorporation assay was used to detect the proliferation of T cells, and the ratio of T cell subsets to T lymphocyte subsets was detected by flow cytometry. Western blot assay was used to detect the B lymphocyte stimulating factor. The expression level and NF- 魏 B activity of BAFF receptor, transmembrane activator and calmodulin ligand interaction molecule (taii) and IL-2 receptor (IL-2R) were observed. The expression of BAFFR or TACI in T cells was inhibited by small interfering RNAs. Results Anti-BAFF antibody could promote the proliferation and activation of T cells in vitro. Interleukin-2, IFN- 纬) and transforming growth factor- 尾 (TGF- 尾) secrete BAFFRTACI. The expression of IL-2R increased and the activity of NF- 魏 B enhanced P0.05N. In vitro administration of 100 渭 g / ml inhibited the proliferation of T lymphocytes induced by anti CD3 antibody (P 0.05). Rh TACI-IgA (100 渭 g / ml) could significantly decrease the level of cytokines induced by anti CD3 antibodies (P0.05 and P0.01). The ratio of CD4 CD69 T cells to CD4 CD154 T cells was significantly inhibited, and the ratio of CD4 CD62L T cells was increased. Moreover, rh TACI-IgA 100 渭 g / ml decreased the expression of IL-2 receptor on T cells. Inhibition of NF- 魏 B activity (P0.05G). Silencing BAFFR or TACI inhibits the proliferation of T cells induced by CD3 antibody. Conclusion Antibody to CD3 is partly produced by BAFF. Activation of BAFFR signal could promote T cell proliferation and activation of rh TACI-Ig and BAFF, and inhibit the expression of BAFF related receptors and the activity of NF- 魏 B. Reduce T cell expression of IL-2 receptor and prevent T cell proliferation and activation.
【作者单位】：安徽医科大学临床药理研究所抗炎免疫药物教育部重点实验室安徽抗炎免疫药物协同创新中心;
【基金】：国家自然科学基金(编号:81173075,81330081,81202541) 安徽省自然科学基金(编号:1208085QH146) 安徽医科大学青年拔尖人才支持计划(2013) 第三批安徽医科大学校级中青年学术技术带头人基金
【分类号】：R392
【正文快照】： 2016-04-06接收抗CD3抗体可以上调T淋巴细胞白细胞介素2受体(interleukine-2 receptor,IL-2R)表达,诱导细胞增殖与活化[1]。活化的T细胞能分泌IL-1、IL-2、IL-6、IL-10和B淋巴细胞刺激因子(B lymphocyte stim-ulator,Bly S or BAFF)等多种细胞因子[2]。BAFF及APRIL水平的异常

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