HBV慢性携带小鼠中CD27和CD11b标记NK细胞亚群的变化

Changes in natural killer cell subsets marked with CD27 and CD11b for HBV carrier mice

目的评价乙型肝炎病毒(HBV)慢性携带小鼠中CD27和CD11b标记自然杀伤(NK)细胞亚群的变化。方法将pAAV-HBVl.2质粒采用高压水动力法尾静脉注入C57BL/6小鼠体内,作为造模组,空载质粒作为对照组;检测不同时间点肝功能和病毒学指标判断HBV质粒慢性携带动物模型构建情况;采用流式细胞术检测脾脏和肝脏中NK细胞及CD11b联合CD27划分的NK细胞亚群的频数;采用Graphpad Prism软件进行统计学分析。结果小鼠HBV持续携带模型构建成功。与对照组相比,HBV慢性携带小鼠的脾脏和肝脏中的NK细胞频数差异均无统计学意义(P值均>0.05)。CD27联合CD11b将NK细胞划分为CD11b+CD27-(CD11b+SP)、CD11b+CD27+(DP)、CD11b-CD27+(D27+SP)和CD11b-CD27-(DN)四个亚群,与对照组相比,HBV慢性携带小鼠的脾脏中四个NK细胞亚群的频数差异均无统计学意义(P值均>0.05)。HBV慢性携带小鼠的肝脏中DN NK细胞亚群的频数与对照组比较显著增多(P<0.001),CD11b+SP细胞亚群的频数显著减少(P<0.05),DP和CD27+SP两个NK细胞亚群频数差异均无统计学意义(P值均>0.05)。结论HBV慢性携带小鼠肝脏NK细胞亚群分布异常,DN NK细胞亚群的频数明显增多,CD11b+SP细胞亚群的频数显著减少。

Objective To evaluate the changes in natural killer cell subsets marked with CD27 and CD11b for HBV carrier mice.Methods 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.Results 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(P>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(P>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(P<0.001);however,the frequency of CD11b+SP cell subsets was significantly decreased(P<0.05).There were no statistical significance in the frequency comparison between NK subgroups of DP and CD27+SP NK cell subsets(P>0.05).Conclusion 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.