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目的 研究SLET细胞是否存在TCR CD3复合物介导的异常生物化学信号转导 ,及其异常与三磷酸肌醇(Inositol1,4 ,5 triphosphate,InsP3)生成量的相关性。方法 用尼龙柱分离肝素化的静脉血得到T细胞并制备其悬液 ,用流式细胞术测定悬液的CD3+ 细胞阳性率 90 %以上。将抗CD3单抗与羊抗鼠IgG相交联刺激T细胞后 ,用粘附细胞仪连续观察10min ,观察T细胞胞内游离Ca2 + 的变化。用流式细胞术检测正常人和SLET细胞上CD3+ 表达的阳性率。用InsP3放射受体试剂盒检测InsP3的生成量。结果 ①正常人和SLET细胞的 [Ca2 + ]i反应的基准值相似 (P =0 .10 5 ) ,SLET细胞的 [Ca2 + ]i反应的高峰值、平台值明显高于正常对照 (P <0 .0 0 1,P <0 .0 0 1)。②正常人和SLET细胞上CD3+ 表达的阳性率没有差异 (P =0 .6 6 5 )。③二者InsP3的生成量无差异 (P =0 .5 37)。结论 SLET细胞TCR CD3介导的 [Ca2 + ]i反应存在异常 ,且与InsP3的生成量无关。
Objective To investigate whether TCR CD3 complex-mediated abnormal biochemical signal transduction exists in SLET cells and its association with abnormal formation of inositol 1,4,4 triphosphate (InsP3). Methods T cells were isolated from the heparinized venous blood by nylon column and the suspension was prepared. The positive rate of CD3 + cells in the suspension was determined to be over 90% by flow cytometry. After anti-CD3 McAb and goat anti-mouse IgG were cross-linked to stimulate T cells, the cells were observed continuously for 10 min by adherent cells to observe the change of intracellular free Ca2 + in T cells. The positive rate of CD3 + expression in normal and SLET cells was detected by flow cytometry. InsP3 production was measured using InsP3 radioimmunoassay kit. Results ① The baseline values of [Ca2 +] i responses in normal and SLET cells were similar (P = 0.105). The peak and plateau values of [Ca2 +] i response in SLET cells were significantly higher than those in normal controls (P < 0 .0 0 1, P <0 .0 0 1). ② There was no difference in the positive rate of CD3 + expression between normal subjects and SLET cells (P = .665). ③ There was no difference in the production of InsP3 (P = 0.537). CONCLUSIONS: TCR CD3-mediated [Ca2 +] i responses in SLET cells are abnormal and are independent of the production of InsP3.