抗人CD134单抗对植物血凝素诱导的外周血单个核细胞穿孔素表达的影响

Effects of anti-human CD134 monoclonal antibody on phytohemagglutinin induced perforin expression in peripheral blood mononudear cells

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 study the concentration-response and time-cotwse effects ot anti-human CD134 monoclonal antibody on the expression of perfofin mRNA and protein in peripheral blood mononuclear cells, and to investigate the possibility of treating autoimmtme diseases by using anti-human CD134 monoclonal antibody. Methods Peripheral blood mononuclear cells were cultured in the presence of phytoheroagglutinin （50 μg/ml） and anti-human CD134 monoclonal antibodies （ 1 μg/ml, 5 μg/ml, 10 μg/ml） for 6 h, 12 h, 24 h and 48 h. Then the expression of perforin mRNA was measured by RT-PCR, and the expression of perforin protein was measured by flow cytometry. Results The expression of perforin mRNA in peripheral blood mononuclear cells decreased in various degrees after treatment with different concentrations of anti-humar. CD134 monoclonal antibodies for different durations and reached the minimum at 24 h at any concentration. In the range of 1 μg/ml-5 μg/ml, anti-human CD134 monoclonal antibody induced downregulation of perforin mRNA of peripheral blood mononuclear cells in a dose-dependent manner （P 〈 0.05）; when the concentration of anti-human CD134 monoclonal antibody exceeded 5μg/ml, the expression of perforin mRNA reached a plateau （ P 〉 0.05 ）. Anti-human CD134 monoclonal antibody also induced the downregulafion of the perforin protein in peripheral blood rnononuclear cells in the same manner. Conclusion Anti-human CD134 monoclonal antibody can inhibit perforin expression on transcription and translation levels. The inhibition reaches its maximum at 24 h and can be saturated. This may be used to treat several autoimmtme diseases.