抗CD3单克隆抗体对食蟹猴免疫功能及相关基因表达的影响

Effects of Anti CD3 Monoclonal Antibody on Immunity and Gene Expression Profiles of Cynomolgus Monkeys

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 investigate the effects of anti CD3 monoclonal antibody （Ant-CD3 McAb） on immune function in cynomolgus monkeys, using the traditional immunotoxicity assessment methods and T cell dependent antibody response （TDAR） test, microarray, etc, and further explore the mechanism of Ant-CD3 McAb. METHODS Eighteen Cynomolgus monkeys were individually dosed with saline, 0. 5 and 2. 5 mg · kg- 1Ant-CD3 McAb for 7 d. The clinical observation, body weight, hematological test, antibody detection, TDAR test, lymphocyte subsets, bone marrow, histopathological examination and whole blood gene expression profile analy- sis were evaluated. RESULTS Ant-CD3 McAb significantly inhibited the immune response to keyhole limpet hemocyanin （KLH） in cynomolgus monkey, and caused the decrease of peripheral white blood cell counts and CD3 ＋ cells, the decrease of lymphocytes in thy- mus cortex. Microarray test showed that the gene function of differently expressed genes mainly involved in ceil proliferation, immune response, cytokine secretion, apoptosis etc. CONCLUSION Ant-CD3 McAb could induce obvious immunosuppression effects in cynomolgus monkeys. Detection of gene expression profile of whole peripheral blood is feasible in immunotoxicity evaluation. The apop- tosis induction and proliferation inhibition of T lymphoeytes might be the main cause of the decreased T lymphocytes. IL-8 might play an important role in the mechanism of Ant-CD3 McAb.