常用实验小鼠感染诺如病毒后外周血免疫指标分析
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(1.中国农业大学动物医学院,北京 100094; 2.中国食品药品检定研究院实验动物资源研究所,北京 102629)

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R-33

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Analysis of peripheral blood immune indexes in commonly used laboratory mice infected with murine norovirus
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(1. College of Veterinary Medicine, China Agricultural University, Beijing 100094, China.2. Laboratory Animal Institute, National Institutes for Food and Drug Control, Beijing 100050)

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    摘要:

    目的 通过小鼠感染MNV 后外周血免疫指标的改变来评估MNV 感染对小鼠免疫功能的影响?方法 选取KM,BALB/ c,BALB/ c-nu,C57BL/6 及NIH 等五个品系小鼠,每个品系分成MNV 感染组和对照组,于感染前(第0 天)和感染后的第7?14?21?28 天从眼周静脉采集抗凝血,流式细胞术测定总T 细胞?CD4+ T 细胞?CD8+ T 细胞?总B 细胞?NK 细胞及NK-T 细胞占总淋巴细胞的百分比及干扰素?白介素?趋化因子等13 种细胞因子的含量,比较感染后各指标总体均值的差异,分析各时间点变化情况?结果 与对照组相比,KM 小鼠感染组总T 细胞?CD4+ T 细胞含量显著增高( P <0. 01),CD4/ CD8 显著增高( P <0. 05),总B 细胞及CXCL10 含量显著降低( P <0. 01);BALB/ c 小鼠CD8+ T 细胞显著降低(P<0. 05),IFN-γ 含量显著增高( P <0. 01);BALB/ c-nu 小鼠各免疫细胞均无统计学差异,TNF-α?CXCL1 及CXCL10 显著增高( P <0. 01),CCL2 显著增高( P <0. 05),IFN-β 显著降低( P <0. 05);C57BL/6 小鼠CD4+( P <0. 05)CD8+ T 细胞( P <0. 01)均显著降低,总B 细胞及IL-10 显著增高( P <0. 05),CCL5 及GM-CSF 显著降低( P <0. 05);NIH 小鼠总T 细胞和CD8+ T 细胞显著增高( P <0. 01),CD4/CD8 显著降低( P <0. 05),总B 细胞显著降低( P <0. 01),细胞因子均无统计学差异?对于有统计学差异的免疫指标,有半数感染第7 天即出现显著差异,大多数第28 天仍存在显著差异?结论 MNV 的感染会影响正常小鼠的细胞免疫和体液免疫应答,而且不同品系的动物影响不同,建议在进行免疫相关的动物实验时选择MNV 阴性小鼠,以避免实验结果出现偏差?

    Abstract:

    Objective To evaluate the effect of murine norovirus (MNV) infection on immune function in miceby assessing the changes in peripheral blood immune parameters. Methods KM, BALB/ c, BALB/ c-nu, C57BL/6, andNIH mice were selected. Each strain was divided into an MNV infection group and a control group. Anticoagulated bloodsamples were collected from the periocular vein plexus before infection (day 0) and at 7, 14, 21, and 28 d post-infection.Flow cytometry was used to determine the percentage of total T cells, CD4+ T cells, CD8+ T cells, total B cells, NK cellsand NK-T cells in the total lymphocytes and the levels of 13 cytokines such as interferon and interleukin, and chemokines.To compare the differences in the mean values of each index after infection, the changes at each time point were analyzed.Results Compared with the control group, the level of total T cells and CD4+ T cells in the infected group of KM micesignificantly increased ( P <0. 01), CD4/ CD8 significantly increased ( P <0. 05), and the total B cell and CXCL10 contentsignificantly decreased ( P <0. 01). The level of CD8+ T cells in infected BALB/ c mice significantly decreased ( P <0. 05),and IFN-γ levels significantly increased ( P <0. 01). There was no significant difference between the levels of immune cellsin infected and control BALB/ c-nu mice, although in the infected BALB/ c-nu mice TNF-α, CXCL1 and CXCL10significantly increased ( P <0. 01), CCL2 significantly increased ( P < 0. 05), and IFN-β significantly decreased ( P <0. 05). CD4+( P <0. 05) and CD8+ T cells ( P <0. 01) in the infected C57BL/6 mice significantly decreased, total B cellsand IL-10 significantly increased ( P <0. 05), and CCL5 and GM-CSF significantly decreased ( P <0. 05). Total T cells andCD8+ T cells in the infected NIH mice significantly increased ( P <0. 01), CD4/ CD8 significantly decreased ( P <0. 05),and total B cells significantly decreased ( P <0. 01), while there were no significant differences in cytokine levels. In termsof statistically significant changes in immune indicators, half of all immune indicators showed significant differences on the7th day of infection, and most still demonstrated significant differences on the 28th day. Conclusions MNV infectionaffects cellular and humoral immune responses in otherwise healthy mice, and different mouse strains demonstrate differenteffects. Researchers should select MNV-negative mice in immunologically relevant animal experiments to avoid bias in the experimental results.

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李晓波,付瑞,王吉,王淑菁,李威,王莎莎,秦骁,黄宗文,贺争鸣,岳秉飞,赵德明.常用实验小鼠感染诺如病毒后外周血免疫指标分析[J].中国比较医学杂志,2019,29(7):67~75.

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  • 收稿日期:2019-02-25
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  • 在线发布日期: 2019-08-01
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