Id2基因过表达对C57BL/6小鼠免疫表型的影响  

Effects of Id2 gene overexpression on immunophenotype of C57BL/6 mice

作　　者：王怡春 任杰 王崎 吴敏娜[2] 贾婷 梁银明 钟根深 WANG Yichun;REN Jie;WANG Qi;WU Minna;JIA Ting;LIANG Yinming;ZHONG Genshen(School of Laboratory Medicine,Xinxiang Medical University,Xinxiang 453003,China)

机构地区：[1]新乡医学院医学检验学院,新乡453003 [2]新乡医学院基础医学院,新乡453003

出　　处：《中国免疫学杂志》2023年第5期904-910,共7页Chinese Journal of Immunology

基　　金：国家自然科学基金项目(81872361,U1904131);河南省科技攻关项目(222102310048)。

摘　　要：目的:建立分化抑制因子2(ID2)过表达和CD45.1,CD45.2嵌合体小鼠模型,探讨ID2过表达(ID2^(o/o) )对C57BL/6小鼠免疫表型的影响。方法:利用基因编辑技术获得ID2^(o/o) C57BL/6模型小鼠,采用PCR、琼脂糖凝胶电泳和Western blot对小鼠进行基因型鉴定。Western blot检测ID2^(o/o) 和WT小鼠脾脏细胞中各种ID蛋白(ID1~ID4)表达情况。流式细胞术绝对定量方法检测WT和ID2^(o/o) 小鼠脾脏和外周血细胞中各免疫细胞数量。构建骨髓1∶1竞争性嵌合体小鼠模型,以同样方法检测CD45.1^(+)和CD45.2^(+)白细胞及相应免疫细胞数量。结果:PCR和Western blot结果表明ID2^(o/o) 小鼠构建成功。在ID2^(o/o) 小鼠脾细胞中,4种ID蛋白的表达量均显著高于WT小鼠。流式细胞分析结果显示,Id2基因过表达使C57BL/6小鼠脾脏和外周血中的中性粒细胞、巨噬细胞、树突状细胞、CD4^(+)T细胞、CD8^(+)T细胞、Treg细胞和NK细胞数量显著增多,该趋势在骨髓嵌合小鼠中也得到进一步验证。结论:Id2基因过表达可增强C57BL/6小鼠体内免疫细胞的分化和发育,提高小鼠各种免疫细胞数量。Objective:To explore effect of differentiation-2(ID2)overexpression(ID2o/o)on immunophenotype of C57BL/6 mice by establishing ID2 overexpression and CD45.1,CD45.2 chimeric mice model.Methods:ID2^(o/o) C57BL/6 model mice were obtained by gene editing technology,and genotypes of mice were identified by PCR,agarose gel electrophoresis and Western blot.Expressions of ID proteins(ID1~ID4)in spleen cells of ID2^(o/o) and WT mice were detected by Western blot.The numbers of various immune cells in spleen and peripheral blood cells of WT and ID2^(o/o) mice were determined by absolute quantitative flow cytometry.Bone marrow 1∶1 competitive chimera mouse model was constructed,and the numbers of CD45.1^(+)and CD45.2^(+)leukocytes and corresponding immune cells were detected by the same method.Results:PCR and Western blot showed that ID2^(o/o) mice were successfully generated.Expression levels of four ID proteins in spleen cells of ID2^(o/o) mice were significantly higher than those of WT mice.Flow cytometry analysis results showed that overexpression of Id2 gene significantly increased the number of neutrophils,macrophages,dendritic cells,CD4^(+)T cells,CD8^(+)T cells,Treg cells and NK cells in spleen and peripheral blood of C57BL/6 mice,and this tendency was further verified in bone marrow chimeric mice.Conclusion:Overexpression of Id2 gene can further support the differentiation and development of immune cells in C57BL/6 mice and significantly increase the number of various immune cells.

关 键 词：ID2 免疫细胞 淋巴细胞 分化 发育 

分 类 号：R392.12[医药卫生—免疫学]