To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type 3(MHV-3) induced chronic viral hepatitis in C3H/Hej mice,ninety C3H/Hej mice were chosen to individually receive 10 plaque...To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type 3(MHV-3) induced chronic viral hepatitis in C3H/Hej mice,ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units(PFU)of MHV-3 intraperitoneally.The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin(HE) staining method from 2 days post MHV-3 infection.The ratios of T cell subsets including CD3+CD4+CD8-,CD3+CD4-CD8+,CD3+CD4-CD8-,CD3+CD4+CD25+,CD3+CD4+CD25-and CD3+ CD4-CD25+ T lymphocyte of total T lymphocytes in blood,spleen and liver were examined at 0,2,4,6,8,10,12,15,20,25,30,40 days post MHV-3 infection by flow cytosorting.We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection.The double negative T cell(DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice,and CD3+CD4+CD8-,CD3+CD4-CD8+,CD3+CD4+CD25-and CD3+CD4-CD25+ T cell ratios decreased accordingly.In conclusion,the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence.Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection.Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.展开更多
Objective To explore the role of MyD88 signaling in MHV-3 virus-mediated fulminant hepatitis. Methods We evaluated liver lesion status, the expression of multiple pro-inflammatory cytokines and HMGB1, the recruitment ...Objective To explore the role of MyD88 signaling in MHV-3 virus-mediated fulminant hepatitis. Methods We evaluated liver lesion status, the expression of multiple pro-inflammatory cytokines and HMGB1, the recruitment of inflammatory ILC3, and mortality in MyD88-/-and WT mice. Results The expression of multiple pro-inflammatory cytokines that recruit inflammatory ILC3 to the liver was severely impaired in MyD88-/-mice resulting in reduced liver pathology, viral replication, and mortality post-infection. Additionally, MHV-3 markedly increased the expression of high-mobility group box 1(HMGB1) in infected hepatocytes/macrophages and induced HMGB1 protein migration from the nucleus to the extracellular milieu, where it activates MyD88-dependent inflammation. Conclusion Our findings indicate that MyD88 exacerbates immunological pathology in experimental viral fulminant hepatitis.展开更多
Recently,the Th17 cells and IL-17 have been shown to play a critical role in the immune-mediated liver injury in hepatitis B,while their functions in acute liver failure have not been well elucidated yet.In this study...Recently,the Th17 cells and IL-17 have been shown to play a critical role in the immune-mediated liver injury in hepatitis B,while their functions in acute liver failure have not been well elucidated yet.In this study,we primarily investigated the role of IL-17 in the development of mouse hepatitis virus strain 3(MHV-3)-induced acute liver failure.IL-17 mRNA levels in liver tissue were quantified by using quantitative real-time polymerase chain reaction,and cytokine IL-17 levels in liver tissue and serum were determined by using ELISA in MHV-3-induced murine fulminant hepatitis model.The IL-17 expression levels on CD4 + T and CD8 + T cells were determined by using flow cytometry.The correlation between IL-17 level and liver injury was studied.Th17 associated cytokines were also investigated by intracellular staining.Our results showed that the IL-17 expression was significantly elevated in the liver and serum of BALB/cJ mice infected with MHV-3.Moreover,a time course study showed that the percentage of both IL-17-producing CD4 + T cells and IL-17-producing CD8 + T cells was increased remarkably in the liver starting from 48 h and peaked at 72 h post-infection.There was a close correlation between hepatic or serum IL-17 concentration and the severity of liver injury defined by ALT level,respectively.Th17 associated cytokines,IL-6,IL-21 and IL-22,were also increased significantly at 72 h post-infection.It was concluded that IL-17 may contribute to the pathogenesis of MHV-3-induced acute liver failure.展开更多
The role of chemokines in murine hepatitis virus strain 3 (MHV-3) induced fulminant hepatic failure (FHF) is not well defined. In this study, we investigated the role of the CXC chemokine receptor 3 (CXCR3)- associate...The role of chemokines in murine hepatitis virus strain 3 (MHV-3) induced fulminant hepatic failure (FHF) is not well defined. In this study, we investigated the role of the CXC chemokine receptor 3 (CXCR3)- associated chemokine [monokine induced by IFN-gamma (Mig/CXCL9) and interferon-gamma-inducible protein 10 (IP-10/CXCL10)] in the recruitment of intrahepatic lymphocytes and subsequent fulminant hepatic failure induced by MHV-3. Balb/cJ mice (6-8 weeks, female) were intraperitioneally injected with 100 PFU MHV-3.The proportions and numbers of T cells and NK cells as well as the expression of CXCR3 on T cells and NK cells in the liver, spleen and blood were analyzed by flow cytometry. The hepatic mRNA level of the CXCR3-associated chemokines (CXCL9 and CXCL10) was detected by realtime PCR. A transwell migration assay was used to assess the chemotactic effect of MHV-3-infected hepatocytes on the splenic lymphocytes. 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. These results suggest that the CXCR3- associated chemokines (CXCL9 and CXCL10) may play animportant role in the recruitment of intrahepatic lymphocytes and subsequent necroinflammation and hepatic failure in MHV-3 infection.展开更多
基金National Nature Science Foundation (30571643, 30672380)Major State Basic Research Development Program of China (2005CB522901,2007CB512904)
文摘To study the contribution of T cell subsets in the pathogenesis of Murine hepatitis virus Type 3(MHV-3) induced chronic viral hepatitis in C3H/Hej mice,ninety C3H/Hej mice were chosen to individually receive 10 plaque forming units(PFU)of MHV-3 intraperitoneally.The changes of virus titer and pathology in liver tissue were examined by standard plaque assay and by the hematoxylin/eosin(HE) staining method from 2 days post MHV-3 infection.The ratios of T cell subsets including CD3+CD4+CD8-,CD3+CD4-CD8+,CD3+CD4-CD8-,CD3+CD4+CD25+,CD3+CD4+CD25-and CD3+ CD4-CD25+ T lymphocyte of total T lymphocytes in blood,spleen and liver were examined at 0,2,4,6,8,10,12,15,20,25,30,40 days post MHV-3 infection by flow cytosorting.We observed that the virus titer raised and showed persistent virus duplications and inflammatory changes in the livers of C3H/Hej mice from 2 days post MHV-3 infection.The double negative T cell(DN Treg cell) and CD4+CD25+ T cell ratios increased significantly from 2 days post MHV-3 infection in C3H/Hej mice,and CD3+CD4+CD8-,CD3+CD4-CD8+,CD3+CD4+CD25-and CD3+CD4-CD25+ T cell ratios decreased accordingly.In conclusion,the changes of virus titer and pathology in the livers of C3H/Hej mice post MHV-3 suggest their contribution to viral persistence.Further characterizations of DN Treg cells are that infection indicates that MHV-3 could induce the chronic inflammation in livers of C3H/Hej mice.The increase of the DN Treg cell and CD4+CD25+ T cell ratios in C3H/Hej mice post MHV-3 infection suggests that DN Treg cells and CD4+CD25+ T cells may both have important suppressive immunomodulation functions in the development of chronic viral hepatitis and have important roles in the virus persistent infection.Further characterizations of DNT cell and CD4+CD25+ T cell are under investigation.
基金Supported by grants from the National Natural Sciences Foundation of China(No.81361120400 and 81222023)
文摘Objective To explore the role of MyD88 signaling in MHV-3 virus-mediated fulminant hepatitis. Methods We evaluated liver lesion status, the expression of multiple pro-inflammatory cytokines and HMGB1, the recruitment of inflammatory ILC3, and mortality in MyD88-/-and WT mice. Results The expression of multiple pro-inflammatory cytokines that recruit inflammatory ILC3 to the liver was severely impaired in MyD88-/-mice resulting in reduced liver pathology, viral replication, and mortality post-infection. Additionally, MHV-3 markedly increased the expression of high-mobility group box 1(HMGB1) in infected hepatocytes/macrophages and induced HMGB1 protein migration from the nucleus to the extracellular milieu, where it activates MyD88-dependent inflammation. Conclusion Our findings indicate that MyD88 exacerbates immunological pathology in experimental viral fulminant hepatitis.
基金This project was supported by grants from National Science Foundation of China Advanced Program (No. 81030007)National Science Foundation of China for Young Scholar (No.81100308/H0318)
文摘Recently,the Th17 cells and IL-17 have been shown to play a critical role in the immune-mediated liver injury in hepatitis B,while their functions in acute liver failure have not been well elucidated yet.In this study,we primarily investigated the role of IL-17 in the development of mouse hepatitis virus strain 3(MHV-3)-induced acute liver failure.IL-17 mRNA levels in liver tissue were quantified by using quantitative real-time polymerase chain reaction,and cytokine IL-17 levels in liver tissue and serum were determined by using ELISA in MHV-3-induced murine fulminant hepatitis model.The IL-17 expression levels on CD4 + T and CD8 + T cells were determined by using flow cytometry.The correlation between IL-17 level and liver injury was studied.Th17 associated cytokines were also investigated by intracellular staining.Our results showed that the IL-17 expression was significantly elevated in the liver and serum of BALB/cJ mice infected with MHV-3.Moreover,a time course study showed that the percentage of both IL-17-producing CD4 + T cells and IL-17-producing CD8 + T cells was increased remarkably in the liver starting from 48 h and peaked at 72 h post-infection.There was a close correlation between hepatic or serum IL-17 concentration and the severity of liver injury defined by ALT level,respectively.Th17 associated cytokines,IL-6,IL-21 and IL-22,were also increased significantly at 72 h post-infection.It was concluded that IL-17 may contribute to the pathogenesis of MHV-3-induced acute liver failure.
基金the National Key Basic Research Program of China (2007CB512900, 2005CB522901, 2005CB 522507)the National Science Fund of China (NSFC 30672380, 30571643)
文摘The role of chemokines in murine hepatitis virus strain 3 (MHV-3) induced fulminant hepatic failure (FHF) is not well defined. In this study, we investigated the role of the CXC chemokine receptor 3 (CXCR3)- associated chemokine [monokine induced by IFN-gamma (Mig/CXCL9) and interferon-gamma-inducible protein 10 (IP-10/CXCL10)] in the recruitment of intrahepatic lymphocytes and subsequent fulminant hepatic failure induced by MHV-3. Balb/cJ mice (6-8 weeks, female) were intraperitioneally injected with 100 PFU MHV-3.The proportions and numbers of T cells and NK cells as well as the expression of CXCR3 on T cells and NK cells in the liver, spleen and blood were analyzed by flow cytometry. The hepatic mRNA level of the CXCR3-associated chemokines (CXCL9 and CXCL10) was detected by realtime PCR. A transwell migration assay was used to assess the chemotactic effect of MHV-3-infected hepatocytes on the splenic lymphocytes. 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. These results suggest that the CXCR3- associated chemokines (CXCL9 and CXCL10) may play animportant role in the recruitment of intrahepatic lymphocytes and subsequent necroinflammation and hepatic failure in MHV-3 infection.
