MEC-1移植法构建慢性淋巴细胞白血病小鼠模型

Preparation of mouse models of chronic lymphocytic leukemia by MEC⁃1 cells transplantation

目的制备并评估慢性淋巴细胞白血病小鼠模型,以期为慢性淋巴细胞白血病的治疗提供临床前模型。方法通过文献查阅及实验确定环磷酰胺(cyclophosphamide,CTX)注射剂量以制备免疫抑制小鼠模型;通过腹腔注射、尾静脉注射将1×10^(7)个MEC-1细胞连续3 d移植到小鼠体内,以制备慢性淋巴细胞白血病小鼠;应用流式细胞术、免疫组化等方法,检测慢性淋巴细胞白血病小鼠外周血白细胞、CD19^(+)CD5^(+)B淋巴细胞变化情况,并分析肝、脾、胸腺、肺、淋巴及骨髓中MEC-1细胞浸润情况。结果确定CTX注射剂量为150 mg/kg注射4 d能够显著降低小鼠免疫系统功能;移植MEC-1细胞后小鼠表现慢性淋巴细胞白血病(chronic lymphocytic leukemia,CLL)特征,其白细胞及B淋巴细胞数量增多、骨髓细胞发生增殖。结论CTX注射剂量为150 mg/kg连续注射4 d,可以使小鼠免疫功能显著下降;成功制备CLL小鼠模型。

Objective To prepare and evaluate a mouse model of chronic lymphocytic leukemia in order to provide a preclinical model for the treatment of chronic lymphocytic leukemia.Methods The cyclophosphamide(CTX)injection dose was determined after a literature review and experiments to prepare an immunosuppressed mouse model;1×10^(7)MEC⁃1 cells were transplanted into mice for three consecutive days by intraperitoneal injection and tail vein injection to prepare mice with chronic lymphocytic leukemia.Flow cytometry,immunohistochemistry,and other method were used to detect changes in peripheral blood leukocytes and CD19^(+)CD5^(+)B lymphocytes in mice with chronic lymphocytic leukemia and to analyze the infiltration of MEC⁃1 cells into the liver,spleen,thymus,lungs,lymph,and bone marrow.Results It was determined that a CTX dose of 150 mg/kg injected for four days could significantly reduce the immune system function of mice.After transplantation with MEC⁃1 cells,mice exhibited chronic lymphocytic leukemia(CLL)characteristics.The number of white blood cells and B lymphocytes increased,and bone marrow cells multiplied.Conclusions A CTX dose of 150 mg/kg injected for four consecutive days significantly reduced the immune function of mice and led to the successful preparation of a mouse model of CLL.