Zhang Q F, Zhang D Z
Institute for Viral Hepatitis, Department of Infectious Diseases, Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), the Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China.
Zhonghua Gan Zang Bing Za Zhi. 2019 Jun 20;27(6):436-439. doi: 10.3760/cma.j.issn.1007-3418.2019.06.009.
To evaluate the changes in natural killer cell subsets marked with CD27 and CD11b for HBV carrier mice. The pAAV-HBVl.2 plasmid was injected into the tail vein of C57BL/6 mice by hydrodynamic injection method to construct HBV-carrier model group and empty vector as the control group. Liver function and virological examination at different time points were used to judge the construction of HBV- plasmid carrier animal model. Flow cytometry was used to detect the frequency of NK cells and CD11b combined with CD27 NK cell subsets in spleen and liver. GraphPad Prism software was used for statistical analysis. HBV-carrier mouse model was successfully constructed. There were no statistically significant difference in NK cell frequencies between spleen and liver of HBV carrier mice (> 0.05), compared to control group. NK cells were divided into four subsets with in combination to CD27 and CD11b: CD11b(+)CD27(-)(CD11b(+)SP), CD11b(+)CD27(+)(DP), CD11b(-)CD27(+)(CD27(+)SP) and CD11b-CD27-(DN). Furthermore, the spleen of HBV-carrier mice had no statistically significant difference (> 0.05) with the frequency of the four NK-cell subsets. The frequency of DN NK cell subsets was significantly increased in the liver of HBV carrier mice than control group (< 0.001); however, the frequency of CD11b(+)SP cell subsets was significantly decreased ( < 0.05).There were no statistical significance in the frequency comparison between NK subgroups of DP and CD27(+)SP NK cell subsets (> 0.05). HBV-carrier mice with abnormal distribution of hepatic NK cell subsets significantly increased and decreased the frequency of DN NK cell subsets and CD11b(+)SP cell subsets.
评估乙肝病毒（HBV）携带小鼠中CD27和CD11b标记的自然杀伤细胞亚群的变化。采用水动力注射法将pAAV-HBV1.2质粒注入C57BL/6小鼠尾静脉，构建HBV携带模型组，以空载体作为对照组。通过不同时间点的肝功能和病毒学检查来判断HBV质粒载体动物模型的构建情况。采用流式细胞术检测脾脏和肝脏中NK细胞以及CD11b与CD27联合标记的NK细胞亚群的频率。使用GraphPad Prism软件进行统计分析。成功构建了HBV携带小鼠模型。与对照组相比，HBV携带小鼠脾脏和肝脏中的NK细胞频率无统计学显著差异（>0.05）。NK细胞根据CD27和CD11b可分为四个亚群：CD11b(+)CD27(-)(CD11b(+)SP)、CD11b(+)CD27(+)(DP)、CD11b(-)CD27(+)(CD27(+)SP)和CD11b-CD27-(DN)。此外，HBV携带小鼠脾脏中这四个NK细胞亚群的频率无统计学显著差异（>0.05）。与对照组相比，HBV携带小鼠肝脏中DN NK细胞亚群的频率显著增加（<0.001）；然而，CD11b(+)SP细胞亚群的频率显著降低（<0.05）。DP和CD27(+)SP NK细胞亚群的NK亚群频率比较无统计学意义（>0.05）。肝脏NK细胞亚群分布异常的HBV携带小鼠中，DN NK细胞亚群和CD11b(+)SP细胞亚群的频率显著增加和降低。
