论文部分内容阅读
目的使用传统免疫毒性评价手段以及T细胞依赖性抗体反应(TDAR)检测、基因芯片等技术,从免疫器官、免疫细胞、免疫分子及基因水平上考察抗CD3单克隆抗体(Ant-CD3 Mc Ab)对食蟹猴免疫功能的影响,探索和验证抗CD3单克隆抗体的作用机制。方法使用18只食蟹猴,每天1次,连续7 d,分别给予生理盐水、0.5和2.5 mg·kg~(-1)抗CD3单克隆抗体,期间进行临床症状观察、体重测定、血液学检查、抗体检测、T细胞依赖性抗体反应检测、淋巴细胞亚群分析、骨髓细胞计数、组织病理学检查和全血基因表达谱分析。结果抗CD3单克隆抗体能明显抑制食蟹猴对外来抗原的免疫应答水平,引起外周血白细胞计数和CD3~+细胞数量不同程度减少,胸腺皮质淋巴细胞数目减少。基因芯片检测发现,抗CD3单克隆抗体引起的主要差异表达基因的基因功能涉及细胞增殖、免疫反应、细胞因子分泌、凋亡等几个方面。结论抗CD3单克隆抗体能够引起食蟹猴出现明显的免疫抑制反应,检测全血基因表达谱的变化在评价免疫毒性方面切实可行。诱导T细胞凋亡和抑制T淋巴细胞增殖可能是抗CD3单克隆抗体引起T淋巴细胞数目减少的主要原因,IL-8可能在抗CD3单克隆抗体的作用机制中具有非常重要的作用。
OBJECTIVE To evaluate anti-CD3 monoclonal antibody (Ant-CD3 Mc Ab) in immune organs, immune cells, immune molecules and genes using traditional immunotoxicity evaluation methods and T cell-dependent antibody reaction (TDAR) On the immune function of cynomolgus monkey to explore and verify the anti-CD3 monoclonal antibody mechanism of action. Methods Eighteen cynomolgus monkeys were used once a day for 7 consecutive days. The rats were given normal saline, 0.5 and 2.5 mg · kg -1 anti-CD3 monoclonal antibody respectively. The clinical symptoms, body weight and hematological examination , Antibody test, T cell-dependent antibody test, lymphocyte subsets analysis, bone marrow cell count, histopathological examination and whole blood gene expression profiling. Results Anti-CD3 monoclonal antibody could significantly inhibit the immune response of cynomolgus monkey to foreign antigen, causing the peripheral blood leukocyte count and the number of CD3 ~ + cells to decrease to some extent, and the number of thymus cortical lymphocytes decreased. Gene chip detection found that anti-CD3 monoclonal antibody-induced major differentially expressed genes involved in gene function of cell proliferation, immune response, cytokine secretion, apoptosis and other aspects. Conclusion Anti-CD3 monoclonal antibody can cause obvious immunosuppression in cynomolgus monkeys. Detecting the changes of gene expression profiles in whole blood is feasible and feasible in evaluating immunotoxicity. Induction of T cell apoptosis and inhibition of T lymphocyte proliferation may be the main reason for the decreased number of T lymphocytes induced by anti-CD3 monoclonal antibodies. IL-8 may play a very important role in the mechanism of anti-CD3 monoclonal antibody.