SAA小鼠模型中T细胞分化及巨噬细胞亚型的研究  

T Cell Differentiation and Macrophage Polarization in the Aplastic Anemia Mouse Model

作　　者：孙静[1] 方芳 谭帅 孙婉玲[1] SUN Jing;FANG Fang;TAN Shuai;SUN Wan-ling(Department of Hematology,Xuanwu Hospital,Capital Medical University,Beijing 100053,China)

机构地区：[1]首都医科大学宣武医院血液科,北京100053

出　　处：《中国生物工程杂志》2023年第12期145-152,共8页China Biotechnology

基　　金：北京市自然科学基金(Z200022)资助项目。

摘　　要：目的:探讨重型再生障碍性贫血(severe aplastic anemia,SAA)小鼠模型中淋巴细胞功能亚群和巨噬细胞亚型的特点。方法:B6D2F1小鼠经过全身照射(total body irradation,TBI)后,接受C57BL/6小鼠的淋巴细胞输注,建立SAA小鼠模型;以仅接受TBI的B6D2F1小鼠为对照组。在造模成功后收集两组小鼠外周血、淋巴结、脾脏和骨髓,通过流式细胞术检测小鼠T淋巴细胞亚群及CD4^(+)和CD8^(+)细胞表型分化情况;流式细胞术和免疫组化观察小鼠骨髓中巨噬细胞比例及其极化状态。结果:与TBI对照组相比,SAA组小鼠在造模的第12~13天出现严重骨髓衰竭,表现为骨髓造血面积明显减少,外周血白细胞、血红蛋白和血小板均显著下降。SAA组小鼠淋巴结、脾脏和外周血中CD8^(+)淋巴细胞比例升高,CD4^(+)/CD8^(+)淋巴细胞比值显著下降。SAA小鼠的CD4^(+)和CD8^(+)淋巴细胞中,CD44^(+)CD62L^(-)效应记忆T细胞增多,而CD44^(-)CD62L^(+)的初始T细胞和CD44^(+)CD62L^(+)中央记忆T细胞比例显著下降。SAA小鼠骨髓中CD11b+F4/80+巨噬细胞增多,且巨噬细胞以CD86^(-)cCD206^(+)和iNOS^(-)cCD206^(+)的M2型巨噬细胞为主。结论:SAA小鼠模型中,CD4^(+)/CD8^(+)淋巴细胞比值显著下降,淋巴细胞主要为表达CD44^(+)CD62L^(-)的效应记忆T细胞;骨髓巨噬细胞增多,且以M2型巨噬细胞为主。Objective:To explore the changes of T cell subsets and macrophage subtypes in severe aplastic anemia(SAA)mouse models.Methods:To induce the SAA mouse models,the B6D2F1 mice first received a 55 Gy total body irradiation(TBI)and then were infused with C57BL/6 lymph node cells at 4×10^(6)/mouse via a tail vein injection,while the B6D2F1 mice receiving TBI alone were used as the control group.The mice were euthanized on day 12⁃13,and their peripheral blood,lymph nodes,spleen,and bone marrow were collected for analysis.T cell subsets and phenotypic differentiation of CD4^(+)and CD8^(+)cells were detected by flow cytometry,and the proportion and polarization of macrophages in bone marrow were detected by flow cytometry and immunohistochemistry.Results:Relative to control mice that received 5 Gy TBI without lymph node cells infusion,SAA mouse models exhibited severe bone marrow failure characterized by a significant reduction in the hematopoietic area of the sternum and decreased levels of white blood cells,hemoglobin,and platelets in peripheral blood 12⁃13 days post⁃radiation.In the SAA group,the frequencies of CD8^(+)T cells increased and the ratio of CD4^(+)/CD8^(+)lymphocytes decreased significantly in lymph nodes,spleen,and peripheral blood.In CD4^(+)and CD8^(+)T cells of SAA mouse models,the percentage of CD44^(+)CD62L^(-)effector memory T cells increased,while the proportion of Naïve T cells and CD44^(+)CD62L^(+)central memory T cells decreased significantly.CD11b+F4/80+macrophages increased in the bone marrow of SAA mouse models,and most macrophages were CD86^(-)cCD206^(+)and iNOS⁃cCD206^(+)M2 macrophages.Conclusions:In the SAA mouse models,the ratio of CD4^(+)/CD8^(+)lymphocytes is significantly decreased,and the lymphocytes are mainly CD44^(+)CD62L^(-)effector memory T cells.Bone marrow macrophages increased and were mainly polarized towards M2 macrophages.

关 键 词：再生障碍性贫血 小鼠模型 巨噬细胞极化 T细胞 

分 类 号：Q343[生物学—遗传学]