抗人B淋巴细胞单克隆抗体清除中国恒河猴体内B淋巴细胞效果观察

B-Lymphocyte Depletion in Chinese Rhesus Macaques with Anti-Human B-Lymphocyte Monoclonal Antibody Rituximab

目的用抗人B淋巴细胞单克隆抗体(利妥昔单抗注射液,Rituximab)通过静脉滴注的方法敲除中国恒河猴体内B淋巴细胞,并观察其敲除效果,为建立B淋巴细胞缺失的恒河猴动物模型提供基础的实验数据。方法选取健康的中国恒河猴两只,静脉滴注抗人B淋巴细胞单克隆抗体,定期采集外周血、腹股沟淋巴结和十二指肠黏膜组织,制备淋巴细胞悬液,应用流式细胞术的方法系统性测定B淋巴细胞及T淋巴细胞亚群的变化。结果静脉滴注利妥昔单抗注射液后24 h,中国恒河猴外周血中B淋巴细胞的缺失即能达到100%,持续约14 d;腹股沟淋巴结中B淋巴细胞在静注后7 d缺失100%;十二指肠黏膜组织中B淋巴细胞在静脉滴注后7 d缺失达到90%,维持28 d。并且在成功敲除B淋巴细胞的情况下,CD4+T及CD8+T淋巴细胞无明显波动,维持在一个稳定的水平。结论利妥昔单抗注射液能有效去除中国恒河猴体内B淋巴细胞,为建立B淋巴细胞缺失动物模型奠定基础。

Objective To observe the effects of rituximab injection,an anti-human B-lymphocyte monoclonal antibody,on eliminating the B lymphocytes from Chinese origin rhesus macaques,and to provide some experimental data for establishing B-cell-depleted animals models and better define the role of B cells in the control of pathogenic simian immunodeficiency virus（SIV） replication in future studies.Methods Two healthy Chinese origin rhesus monkeys were selected and infused with rituximab via the tail vein by intravenous drip.Peripheral blood,lymph nodes and duodenum biopsy samples were collected at different time points after injection.Lymphocytes were isolated from all the samples,stained and analyzed by FACS.Results The B lymphocytes in peripheral blood of rhesus monkeys were completely depleted 24 hours after the rituximab infusion and lasted for 14 days,while 100% depletion in the inguinal lymph nodes and 90% depletion in the duodenal mucosa at 7 days after infusion.At the same time,CD4＋ T and CD8＋ T-cells in peripheral blood maintained at a stable level without significant fluctuation.Conclusions B lymphocytes can be effectively eliminated from various tissues in Chinese origin rhesus macaques by rituximab injection in vivo.