Thapsigargin及EGTA对SLE T细胞TCR/CD_3复合物介导的[Ca^(2+)]i反应的影响

Study on Effect on TCR/CD_3 Complex-mediated [Ca^(++)]i Responses in T Cells From Patients With SLE by Thapsigargin and EGTA

目的 证实SLE患者T细胞功能异常是否与TCR/CD3 复合物介导的 [Ca2 + ]反应异常有关 ,以及 [Ca2 + ]i反应异常的原因。方法 用CD3 单抗与羊抗鼠二抗IgG相交联刺激T细胞并用Thapsigargin和EGTA干预后 ,分别粘附细胞仪连续观察 10minT细胞 [Ca2 + ]i的变化 ,并评价 [Ca2 + ]i反应与InsP3 生成量的相关性。结果 正常人和SLE患者T细胞 [Ca2 + ]i反应的基准值相似(P =0 10 5 ) ;SLE患者T细胞的 [Ca2 + ]i反应高峰值、平台值明显高于正常对照 (P <0 0 0 1,P <0 0 0 1) ;加入Thapsigargin后二者[Ca2 + ]i反应无显著差异 ,而加入EGTA后二者 [Ca2 + ]i反应有显著差异 ;二者的T细胞InsP3 生成量无差异 (P =0 5 3 7)。结论 SLE患者T细胞TCR/CD3 介导的 [Ca2 + ]i反应存在异常 ,这种 [Ca2 + ]i反应异常是储存库内Ca2 +

Objective To demonstrate that SLE T cells primary function disordor was related to abnormal [Ca 2+ ]i responses,and to investigate the reason of abnormal [Ca 2+ ]i responses.Methods After cross-linking of anti-CD 3 mAb to sheep anti-mouse IgG and stimulating T cells,the changes of free calciumion [Ca 2+ ]i within T cells under interference of Thapsigarain and EGTA was observed respectively successively for 10 minutes by an adhesion cytometry.The relation between [Ca 2+ ]i responses in SLE T cells and InsP 3 levels was evaluated .Results The basic [Ca 2+ ]i response in T cells from SLE patients was similar to that from normal control(P=0 105),peak and plateau [Ca 2+ ]i responses were significantly higher in the group of SLE patients(P<0 001,P<0 001),[Ca 2+ ]i responses were similar in both individuals after addition of Thapsigargin,[Ca 2+ ]i responses of T cells from SLE patients in the presence of EGTA was higher,InsP 3 generation was similar in both individuals(P=0 537).Conclusion TCR/CD 3 [Ca 2+ ]i responses in T cells from SLE is abnormal,the abnormality might result from increased release of Ca 2+ from intracellular stores.