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目的探讨LIGHT-HVEM/LTbR信号通路缺陷对T细胞应答的影响。方法以LIGHTKO小鼠为动物模型,制备脾脏单细胞悬液,加入抗CD3mAb和抗CD28mAb刺激。通过3H-TdR掺入法检测了细胞的增殖能力、ELISA和FACS分别检测了细胞培养上清和胞内IFN-gamma和IL-10的水平,并探讨了这些细胞对外源性rIL-2的反应性。结果与野生型的C57BL/6小鼠相比,LIGHTKO小鼠的脾细胞表现为低下的增殖活性(P<0.05);细胞培养上清中Th1细胞因子IFN-gamma的水平显著下降(P<0.05);与ELISA结果一致,FACS胞内染色结果表明,IFN-gamma阳性T细胞的百分率显著下降,而LIGHTKO小鼠脾细胞IL-10的产生却显著高于野生型的对照小鼠(P<0.01)。进一步的研究表明LIGHTKO小鼠这种缺陷的IFN-gamma产生主要与CD8+T细胞亚群有关,而CD4+T细胞亚群IFN-gamma的产生是正常的。外源性rIL-2的加入可使LIGHTKO小鼠T细胞恢复正常的增殖能力和IFN-gamma的产生。结论 LIGHT-HVEM/LTbR信号通路缺陷导致T细胞的失能。
Objective To investigate the effect of LIGHT-HVEM / LTbR signaling pathway on T cell response. Methods LIGHTKO mice were used as animal models to prepare single cell suspension of spleen, and then stimulated by anti-CD3 mAb and anti-CD28 mAb. The proliferative ability of cells was detected by 3H-TdR incorporation. The levels of IFN-gamma and IL-10 in supernatants and intracellular cells were detected by ELISA and FACS respectively. The reactivity of these cells to exogenous rIL-2 . Results Compared with the wild type C57BL / 6 mice, the spleen cells of LIGHTKO mice showed low proliferative activity (P <0.05), and the levels of Th1 cytokine IFN-gamma in the cell culture supernatant decreased significantly (P <0.05 ). Consistent with the results of ELISA, intracellular staining of FACS showed that the percentage of IFN-gamma positive T cells was significantly decreased, while the production of IL-10 in LIGHTKO mice spleen cells was significantly higher than that of wild-type control mice (P <0.01 ). Further studies showed that IFN-gamma production in LIGHTKO mice was mainly associated with CD8 + T cell subsets, whereas CD4 + T cell subsets IFN-gamma production was normal. The addition of exogenous rIL-2 restored normal proliferation and IFN-gamma production in LIGHTKO mouse T cells. Conclusions Defects of LIGHT-HVEM / LTbR signaling pathway result in T cell dysfunction.