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目的观察不同浓度抗人CD134单抗对外周血单个核细胞(PBMC)穿孔素表达的影响及时间效应,旨在探讨抗人CD134单抗治疗自身免疫性疾病的可能机制。方法在植物血凝素诱导下,分别以1μg/ml、5μg/ml和10μg/ml的抗人CD134单抗作用于人PBMC,于6 h、12 h、24 h、48 h用反转录-聚合酶链反应(RT-PCR)检测穿孔素mRNA表达,流式细胞仪检测穿孔素蛋白表达。结果不同浓度的CD134单抗作用不同时间后,健康成人PBMC穿孔素mRNA和蛋白的表达均有不同程度的下调,且在24 h达最低值。当CD134单抗≤5μg／ml时,随着单抗浓度的增加,穿孔素mRNA明显下调,其差异有统计学意义,当CD134单抗浓度>5μg／ml时,穿孔素mRNA不再下调(P>0．05);穿孔素蛋白表达也有类似的变化。结论抗人CD134单抗可以在转录水平和翻译水平上抑制穿孔素的表达,这种作用在24 h达到高峰且会饱和,这可能被用于治疗某些自身免疫性疾病。
Objective To investigate the effect of different concentrations of anti-human CD134 monoclonal antibody on perforin expression in peripheral blood mononuclear cells (PBMCs) and the time effect of anti-human CD134 monoclonal antibody in the treatment of autoimmune diseases. Methods Human PBMCs were induced with 1μg / ml, 5μg / ml and 10μg / ml of anti-human CD134 monoclonal antibody induced by phytohemagglutinin, respectively, and then transfected into human PBMCs at 6, 12, 24, Perforin mRNA expression was detected by polymerase chain reaction (RT-PCR) and perforin expression by flow cytometry. Results The expressions of perforin mRNA and protein in PBMC of healthy adults were down-regulated to different extents at different concentrations of CD134 mAb at different times, reaching the lowest value at 24 h. When CD134 monoclonal antibody was lower than or equal to 5μg / ml, perforin mRNA was significantly down-regulated with the increase of monoclonal antibody concentration, and the difference was statistically significant. When CD134 mAb concentration was> 5μg / ml, perforin mRNA was no longer down-regulated > 0.05). Perforin expression also had similar changes. CONCLUSION: Anti-human CD134 mAb inhibits perforin expression at the transcriptional and translational levels. This effect peaked at 24 h and saturates, which may be used to treat certain autoimmune diseases.