淋巴细胞功能相关抗原1共刺激作用在活动性狼疮肾炎中的作用

目的探讨淋巴细胞功能相关抗原(LFA)1及其信号分子在狼疮肾炎发病机制中的作用。方法 利用免疫沉淀、免疫印迹和RTllPCR技术检测了LFA-1共刺激对19例活动性狼疮肾炎(LN)患者外周血单个核细胞(PBMC)P13-K磷酸化产物、IL-10 mRNA表达的影响。以14例健康献血员作为对照。结果 在CD3 mAb30 ng/ml存在的情况下,LFA-1mAb共刺激明显提高了活动性LN患者PBMC P13-K磷酸化产物表达(P<0.01),伴有IL-10 mRNA表达和蛋白含量增加(P<0.01),抗lFA-1抗体(5μmol/L)加入后明显抑制了P13-K磷酸化产物表达(P<0.01)。在CD3 mAb和LFA-1mAb协同刺激的情况下,P13-K特异抑制剂Wortamannin的加入明显抑制了LN患者PBMCIL-10 mRNA水平和蛋白含量(P<0.01)。结论 LFA-1作为共刺激分子过度活化P13-K可能介导了活动性狼疮肾炎IL-L 10的高效表达。

Investigation of signaling pathway by LFA-1 costimulation in PBMC of active lupus nephritis

Objective To study the effect of LFA-1 costimulation and its signal molecule in peripheral blood monoiiuclear cell (PBMC) of active lupus nephritis (LN). Methods Nineteen patients with active LN and 14 controls were selected for this study. PBMCs were obtained with Ficol density gradient centrifugation. P13-K phosphoryjated products were detected by immunoprecipitation and Western blot. RT-PCR was used to examine interleukin-lOmRNA expression, and the content of IL-10 protein from PBMCs was measured by ELISA. Results The level of P13-K phosphorylation and interleukin-lOmRNA remarkedly enhanced by co-stimulation with anti-LFA-1 mAb and suboptimal doses of anti-CD3 mAb in PBMC of active LN ( P < 0. 01 ), and IL-10 protein content increased significanlly as well( P < 0. 01). Such effects of LFA-1 costimulation were inhibited by Neutrlizing anti-LFA-1 antibody and PY294002, one of specific inhibitors of P13-K. Conclusion Abnormal signaling pathway by LFA-1 costimulation may contribute to ihe overe.xpresiou of IL-10 in active lupus nephritis.