SLE 患者T细胞TCR/CD3介导[Ca2+]I反应与InsP3生成量的相关性

目的 :研究SLE患者T细胞功能异常是否与TCR/CD3复合物介导的生物化学信号传导异常有关 ,以及这种信号转导异常与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CD3+细胞的阳性率。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SLE患者T细胞上CD3+ 表达的阳性率是否有差异。用InsP3 放射受体试剂盒检测InsP3 的生成量。结果 :①用尼龙柱吸附除去B细胞得到T细胞悬液 ,用流式细胞术检测CD3+ 细胞的阳性率占90 %以上。②正常人和SLE患者T细胞的 [Ca2 + ]i反应的基准值相似 (P =0 .10 5 ) ,SLE患者T细胞的 [Ca2 + ]i反应的高峰值、平台值明显高于正常对照 (P <0 .0 0 1)。③正常人和SLE患者T细胞上CD3+ 表达的阳性率没有差异 (P =0 .6 6 5 )。④二者InsP3 的生成量无差异 (P =0 .5 37)。结论 :SLE患者T细胞TCR/CD3介导的 [Ca2 + ]i反应存在异常 ,这种异常与InsP3 的生成量无关

Study on corelation between InsP3 generation and TCR/ CD3 complex-mediated [ Ca2+]i responses in T cells from SLE patients

AIM: To demonstrate whether function disorder of T cells from SLE patients was relative to abnormal biochemical pathways mediated by TCR/ CD3 complex and whether this abnormality was relative to the InsP(3) production. METHODS: Human T cells were isolated through Nylon-columns from heparinized peripheral blood from SLE patients and control individuals. Percentage of CD3(+) T cells was detected by flow cytometry. After cross-linking of anti-CD3 mAbs to sheep anti-mouse IgG and stimulating T cells, the changes of free calcium ion within T cells was observed successively for 10 minutes by an adhesion cytometry. Flow cytometry was used to detect the difference in positive rate between normal individual's and SLE patient's T cells. Level of InsP(3) was detected by a radioreceptor assay kit. RESULTS: (1)Flow cytometry analysis showed that CD3(+) T cells obtained through nylon column accounted for more than 90%.(2)The base line recordings of [Ca(2+)]i response from SLE patients were similar to that from normal control (P=0.105). Peak and plateau [Ca(2+)]i response in T cells of SLE patients were significantly higher than that in the control group (P<0.001). (3)The percentage of the CD3(+) T cells was similar in both individuals (P=0.665).(4)No differences in the anti-CD3 mAb-mediated InsP(3) generation were found between the two groups (P=0.537). CONCLUSION: TCR/ CD3-mediated [Ca(2+)]i responses in T cells from SLE patients is abnormal, and the abnormality has no relation to InsP(3) generation.