趋化因子受体CXCR3与其配体在小鼠暴发性肝炎发病中免疫学机制研究

目的 探讨趋化因子受体CXCR3与其配体(CXCL9/Mig,CXCL10/IP-10)在小鼠暴发性肝炎淋巴细胞迁移和急性肝衰竭中的作用.方法 6～8周龄雌性BALB/cJ小鼠腹腔注射100 PFU 3型鼠肝炎病毒(MHV-3),采用流式细胞术检测感染MHV-3后的BALB/cJ小鼠肝脏、脾脏和外周血T细胞和NK细胞的比例、数量以及其表面趋化因子受体CXCR3的表达频率.实时定量PCR技术检测感染MHV-3后的BALB/cJ小鼠肝内趋化因子CXCL9和CXCL10 mRNA的表达水平.Transwell细胞迁移试验评估病毒感染的肝细胞及CXCL10对脾脏淋巴细胞的趋化作用.结果 BALB/cJ小鼠感染MHV-3后,肝脏T细胞和NK细胞的数量及CXCR3的表达频率均显著增加,然而在脾脏和外周血均显著减少.实时定量PCR检测证实,感染MHV-3 48 h后,肝内趋化因子CXCL9和CXCL10 mRNA的表达比感染前分别上升了15.6和98.8倍.体外Transwell试验表明,病毒感染的肝细胞及重组CXCL10/IP-10蛋白对脾脏T细胞和NK细胞具有明显的趋化作用,并且这种趋化作用能被抗-CXCL10抗体显著阻断.结论 趋化因子受体CXCR3与其配体(CXCL9和CXCL10),尤其是CXCL10的相互作用在小鼠暴发性肝炎肝内淋巴细胞的募集及随后的坏死性炎症和急性肝衰竭中可能发挥着重要作用.

Abstract：

Objective To investigate the role of the chemokine receptor CXCR3 and its ligands in the migration of lymphocytes and acute hepatic failure. Methods BALB/cJ mice (6-8 weeks, female) were intraperitoneally injected with 100 PFU mouse hepatitis virus-3(MHV-3). The proportions and numbers of T cells and NK cells in liver, spleen, and blood as well as the expression of CXCR3 in T cells, and NK cells post MHV-3 infection was analyzed by flow cytometry. The hepatic mRNA level of the CXCR3-associated chemokines(CXCL9 and CXCL10) was detected by real-time PCR. A transwell migration assay was used to assess the chemotactic effect of MHV-3-infected hepatocytes and CXCL10 on the splenic lymphocytes. Results Following MHV-3 infection, the number of hepatic NK cells and T cells and the frequencies of hepatic NK cells and T cells expressing CXCR3 increased markedly; however, in the spleen and peripheral blood, they both decreased significantly. Moreover, the hepatic mRNAs levels of CXCL9 and CXCL10 were significantly elevated post infection. The transwell migration assay demonstrated that MHV-3-infected hepatocytes have the capacity to attract and recruit the splenic NK cells and T cells, and CXCL10 plays a key role in lymphocyte mobilization from the spleen. Conclusion Interactions between CXCR3 and its ligands (CXCL9 and CXCL10),especially CXCL10 may play a key role in the recruitment of intrahepatic lymphocytes and subsequent necroinflammation and acute hepatic failure in MHV-3 infection.

Preliminary study on immunologic mechanism of chemokine receptor CXCR3 and its ligands involved in mouse fulminant hepatitis

Objective To investigate the role of the chemokine receptor CXCR3 and its ligands in the migration of lymphocytes and acute hepatic failure. Methods BALB/cJ mice (6-8 weeks, female) were intraperitoneally injected with 100 PFU mouse hepatitis virus-3(MHV-3). The proportions and numbers of T cells and NK cells in liver, spleen, and blood as well as the expression of CXCR3 in T cells, and NK cells post MHV-3 infection was analyzed by flow cytometry. The hepatic mRNA level of the CXCR3-associated chemokines(CXCL9 and CXCL10) was detected by real-time PCR. A transwell migration assay was used to assess the chemotactic effect of MHV-3-infected hepatocytes and CXCL10 on the splenic lymphocytes. Results Following MHV-3 infection, the number of hepatic NK cells and T cells and the frequencies of hepatic NK cells and T cells expressing CXCR3 increased markedly; however, in the spleen and peripheral blood, they both decreased significantly. Moreover, the hepatic mRNAs levels of CXCL9 and CXCL10 were significantly elevated post infection. The transwell migration assay demonstrated that MHV-3-infected hepatocytes have the capacity to attract and recruit the splenic NK cells and T cells, and CXCL10 plays a key role in lymphocyte mobilization from the spleen. Conclusion Interactions between CXCR3 and its ligands (CXCL9 and CXCL10),especially CXCL10 may play a key role in the recruitment of intrahepatic lymphocytes and subsequent necroinflammation and acute hepatic failure in MHV-3 infection.