CD69+NK Cells Contribute to the Murine Hepatitis Virus Strain 3-Induced Murine Hepatitis
CD69+NK Cells Contribute to the Murine Hepatitis Virus Strain 3-Induced Murine Hepatitis
复制标题
CD69 NK 细胞导致鼠肝炎病毒株 3 诱导的鼠肝炎
DOI：
10.1007/s11596-013-1150-7
复制
发表时间：
2013-08-01
影响因子：
--
通讯作者：
Ning, Qin
中科院分区：
文献类型：
--
作者：
Ding, Lin;Chen, Tao;Ning, Qin
肝脏CD 69+自然杀伤（NK）细胞在病毒诱导的严重肝损伤和随后的肝功能衰竭中的作用尚未明确。本研究利用小鼠肝炎病毒3型（MHV-3）诱导的暴发性肝衰竭（FHF）小鼠模型，研究肝脏CD 69 + NK细胞在FHF发生发展中的作用。采用流式细胞术检测肝、脾、骨髓及外周血NK细胞CD 69的表达。研究肝NK细胞CD 69表达水平与肝损伤的关系。采用流式细胞术检测CD 69 + NK细胞和CD 69-NK细胞表面功能标志物（CD 107 a）、活化和抑制受体（NKG 2D和NKG 2A）的表达。还通过使用细胞内染色检查促炎细胞因子（IL-9、IFN-γ和TNF-α）。BALB/cJ小鼠感染MHV-3后，肝脏中CD 69 + NK细胞数量显著增加，72 h达高峰。在脾脏、骨髓和外周血中也观察到类似的变化。同时，肝脏NK细胞CD 69表达与血清ALT、AST水平高度相关。在感染后48-72 h，肝脏CD 69 + NK细胞中CD 107 a和NKG 2D的表达以及TNF-α、IFN-γ和IL-9的产生均显著上调。相比之下，肝脏CD 69-NK细胞中NKG 2A表达增加，但CD 69 + NK细胞中未增加。这些结果表明，肝脏CD 69 + NK细胞通过增强NK细胞的脱颗粒和细胞毒能力以及增加促炎细胞因子的产生在FHF的发病机制中起关键作用。
The role of hepatic CD69+ natural killer (NK) cells in virus-induced severe liver injury and subsequent hepatic failure is not well defined. In this study, a mouse model of fulminant liver failure (FHF) induced by murine hepatitis virus strain 3 (MHV-3) was used to study the role of hepatic CD69+ NK cells in the development of FHF. The CD69 expression in NK cells in the liver, spleen, bone marrow and peripheral blood was detected by using flow cytometry. The correlation between the CD69 level in hepatic NK cells and liver injury was studied. The functional marker (CD107a), and activating and inhibitory receptor (NKG2D and NKG2A) expressed on CD69+ NK cells and CD69-NK cells were detected by using flow cytometry. Pro-inflammatory cytokines (IL-9, IFN-gamma and TNF-alpha) were also examined by using intracellular staining. After MHV-3 infection, the number of CD69+ NK cells in the liver of BALB/cJ mice was increased markedly and peaked at 72 h post-infection. Similar changes were also observed in the spleen, bone marrow and peripheral blood. Meanwhile, the CD69 expression in hepatic NK cells was highly correlated with the serum level of ALT and AST. The expression of CD107a and NKG2D, as well as the production of TNF-alpha, IFN-gamma and IL-9 in hepatic CD69+ NK cells was all significantly up-regulated during 48-72 h post-infection. In contrast, the NKG2A expression was increased in hepatic CD69-NK cells but not in CD69+ NK cells. These results suggested that hepatic CD69+ NK cells play a pivotal role in the pathogenesis of FHF by enhancing degranulation and cytotoxic ability of NK cells and increasing the production of pro-inflammatory cytokines.