小鼠乳腺表达抗PD-1抗体对其脾脏T细胞的影响  

Effect of expressing of anti-PD-1 antibody in mouse mammary gland on spleen T cells in transgenic mice

作　　者：张子寒 龚桂花 谢丽萍 胡又佳 ZHANG Zihan;GONG Guihua;XIE Liping;HU Youjia(China State Institute of Pharmaceutical Industry,Shanghai 201203,China)

机构地区：[1]中国医药工业研究总院,上海201203

出　　处：《生物工程学报》2023年第1期231-247,共17页Chinese Journal of Biotechnology

基　　金：上海市自然科学基金(19ZR1454300)。

摘　　要：为从细胞水平研究小鼠乳腺表达抗PD-1抗体对转基因鼠脾脏T细胞表面抗原蛋白、细胞因子表达、脾脏CD4+T细胞增殖以及增殖相关通路的影响,将8周龄未经历过怀孕的和18周龄经历过哺乳的表达抗人PD-1抗体的转基因小鼠分成两组,每组以转基因阴性鼠为对照,提取脾脏淋巴细胞,检测脾脏淋巴细胞的变化。与转基因阴性小鼠相比,乳腺表达抗PD-1抗体的转基因小鼠的免疫系统中的脾脏T细胞的效应T细胞比例上升,Treg细胞比例下降,CD4^(+)T细胞表达的IFN-γ、IL-17以及IL-2有不同程度的增加。IL-4、IL-10以及TGF-β都没有发生变化。与T细胞刺激相关的一些细胞表面的蛋白分子也没有引起变化。转基因阳性鼠和转基因阴性鼠中T细胞增殖没有显著性差异,转基因阳性鼠中PI3K/Akt/mTOR和Ras/MEK/ERK这两条通路上的磷酸化蛋白只有部分表达上调,整个通路没有完全上调。结果表明,转基因小鼠作为表达抗PD-1抗体这类免疫系统相关单克隆抗体的宿主是可行的。This study aims to investigate the effect of anti-PD-1 antibody expressed in mouse mammary gland on the surface antigen protein of spleen T cells,cytokine expression,spleen CD4+T cell proliferation and proliferation related pathways of transgenic mice at the cellular level.Transgenic mice expressing anti-human PD-1 antibody at 8 weeks of age without pregnancy and 18 weeks of age with lactation were divided into two groups,with transgenic negative mice in each group as the control.Spleen lymphocytes were extracted and the changes of spleen lymphocytes were detected.Compared with transgenic negative mice,the proportion of effector T cells of spleen T cells in the immune system of transgenic mice with anti-PD-1 antibody expressed in breast increased,the proportion of Treg cells decreased,and the IFN-γ,IL-17 and IL-2 expressed in CD4^(+)T cells increased in varying degrees.Moreover,IL-4,IL-10 and TGF-βin CD4^(+)T cells did not change,nor did some cell surface protein molecules related to T cell stimulate.There was no significant difference in T cell proliferation between transgenic positive and transgenic negative mice.In transgenic positive mice,the expression of phosphorylated proteins in PI3K/Akt/mTOR and RAS/MEK/ERK pathways were partially up-regulated,but the whole pathway was not completely up-regulated.Therefore,it is feasible to use transgenic mice as host to express monoclonal antibodies related to immune system such as anti-PD-1 antibody.

关 键 词：转基因小鼠 抗PD-1抗体 脾脏T细胞 细胞因子 细胞表面抗原蛋白 信号通路 

分 类 号：R730.51[医药卫生—肿瘤]