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 McAb)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。
Abstract
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 analysis 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 thymus cortex. Microarray test showed that the gene function of differently expressed genes mainly involved in cell 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 apoptosis induction and proliferation inhibition of T lymphocytes might be the main cause of the decreased T lymphocytes. IL-8 might play an important role in the mechanism of Ant-CD3 McAb.
关键词
抗CD3单克隆抗体 /
食蟹猴 /
免疫抑制 /
外周血 /
基因表达谱
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Key words
Ant-CD3 McAb /
cynomolgus monkey /
immunosuppression /
peripheral blood /
gene expression profile
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中图分类号：
R992
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参考文献
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脚注
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基金
科技部“十二五”重大新药创制课题资助项目(2012ZX09302001);科技部“十二五”新药研究开发关键技术研究课题资助项目(2012ZX09505001-004)
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