| Objectives Observation the changes of LAG-3 expression levels on peripheral blood T lymphocytes in the HBV infection patients with different natural history,to evaluate the relationship between immunosuppressive molecules LAG-3 in the changes in the expression of T cells and the T cells’ function,the viral load,the degree of liver damage and the antiviral therapeutic effect with different phase of the natural course of HBV infection patients.Methods In the current study,30 IT phase,30 IC phase,30 LR phase of chronic HBV-infected patients and 30 health people were enrolled.The LAG-3 expression levels on peripheral blood T lymphocytes were measured by flow cytometry.Biochemical analyses,immunoassay and PCR were used to detect the liver function,serological markers of HBV and HBV DNA viral copies,respectively.Mann-Whitney U test was used to compare the expression levels of LAG-3 among groups;Spearman correlation analysis was performed to analyze the relationships among the serum levels of ALT,HBeAg,HBV DNA and the positive expression rate of LAG-3.Then,10 IT phase,10 IC phase,10 LR phase of chronic HBV-infected patients were enrolled.After the stimulation of HBsAg,HBcAg and HBeAg,the LAG-3,HLA-DR,IL-4,IFN-γ and Granzyme B expression levels on CD8+ T lymphocytes were measured by flow cytometry.Mann-Whitney U test was used to compare the expression levels of LAG-3,HLA-DR,IL-4,IFN-γ and Granzyme B among the groups.The last,5 IT phase of chronic HBV-infected patients were enrolled.After blocking of LAG-3,Wilcoxon paired rank sum test analysis was used to compare the changes of CD8+T cells after blocking LAG-3,HLA-DR,IL-4,IFN-γ,Granzyme B.Results A statistically higher ratio of LAG-3+CD8+T cells was confirmed in chronic HBV-infected patients than in health controls.Patients in the IT phase of chronic HBV-infected had even higher ratio of LAG-3+CD8+T cells than those in IC phase,LR phase and health controls.The ratio of peripheral blood LAG-3+CD8+T cells in IC phase of chronic HBV-infected patients were negatively correlated with the serum ALT level.The ratio of peripheral blood LAG-3+CD8+T cells in IT phase of chronic HBV-infected patients were positively correlated with the HBeAg level.However,there was no correlation between the ratio of peripheral blood LAG-3+CD4+T cells or LAG-3+CD8+T cells and HBV DNA viral copies in chronic HBV-infected patients.After the stimulation of HBsAg,HBc Ag and HBeAg,patients in the IT phase of chronic HBV-infected had even higher ratio of LAG-3+CD8+T cellsthan those in IC phase and LR phase,but had lower ration of Granzyme-B+ CD8+T cells than the other two phases.Patients in the IC phase of chronic HBV-infected had even higher ratio of HLA-DR+CD8+T cells,IL-4+CD8+T cells,IFN-γ+CD8+T cellsthan those in IT phase.And the last,after blocking of LAG-3,the expression of LAG-3+CD8+T lymphocytes was significantly lower than that in the non blocking group.By contrast,statistically higher ratios of HLA-DR+CD8+T cells,IL-4+CD8+T cells,IFN-γ+CD8+T cells,Granzyme-B+ CD8+T cells was confirmed in blocking group than in non blocking group.Conclusions LAG-3,an immunoinhibitory receptor,is highly expressed in peripheral blood CD8+T cells in chronic HBV-infected patients,especially in patients experiencing IT phase is one of the vital immune factors leading to the dysfunction of CD8+T cells and the continuous replication of HBV in patients with chronic HBV infection.Blocking or inhibiting the expression or signaling pathway of LAG-3 on T cells will contribute to the complete elimination of HBV in the patients with chronic HBV infection. |
