摘要
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。
ObjectiveTo demonstrate whether abnormal biochemical p at hways mediated by TCR/CD3 complex exists in SLE T cells and to further investig ate corelation between the abnormality and InsP 3 generation.Methods Human T cells were isolated through Nylon columns from heparinized peri pheral blood from SLE patients and control individuals. Percentage of CD3 + T cells was more than 90% detected by flow cytometry. After cross linking of anti CD3 mAbs to sheep anti mouse IgG and stimulating T cells, the changes of fr ee calcium ion within T cells was observed successively for 10 minutes by an adhes ion cytometry. Percentage of CD3 + in SLE and control T cells was detected by f low cytometry. InsP 3 levels was detected by a radioreceptor assay kit. Results ① Basal [Ca 2+ ]i responses of SLE and control T cells w ere similar (P=0.105). ② Peak and plateau [Ca 2+ ]i responses in SLE T cells were significantly higher than that in control T cells (P<0.001, P<0.001). ③ The percentage of the CD3 + T cells was similar between the two groups (P=0.665).④ No difference in the anti CD3 mAb mediated Ins P 3 generation was found between the two groups (P=0.537). Conclusion TCR/ CD3 mediated [Ca 2+ ]i responses in SLE T cells is abnormal, and the abnormality has no relation to InsP 3 generation.
出处
《免疫学杂志》
CAS
CSCD
北大核心
2003年第1期56-58,共3页
Immunological Journal