| Hepatitis B is an infectious disease in the whole world. It has been reported thatabout20million people worldwide have been infected with hepatitis B virus (HBV), ofwhich about350million people are HBV carriers. There are millions of people who diedfrom HBV infection caused by the end-stage liver disease, such as cirrhosis, liver failureand liver cancer every year. The specific immune mechanisms of HBV infection are stillunclear. In the recent years, several studies have demonstrated that multiple inhibitoryreceptors were expressed on the surface of T cells during chronic virus infection, whichweaken the function of the virus-specific CD8+T cells. The another study revealed that T-bet expression negatively correlates with the expression of inhibitory receptors such asPD-1,LAG-3in the mice infected by lymphocytic choriomeningitis virus. We will detectthe levels of IFN-γ, T-bet, PD-1, LAG-3, CTLA-4on the CD8+T cells in the HBVinfection patients with different clinical manifestations and analyze the relationship amongthe factors mentioned above. Moreover, we will analyze the relationship between thesefactors and the serology, virology, biochemical parameters of the HBV infection patientswith different clinical manifestations respectively. The aim of the study is to provideexperimental data for the pathogenesis of the HBV infection, and to find out new therapytargets for the chronic HBV infection.Aims:To analyze the differences and relationship of expression lever of IFN-γ, T-bet, PD-1,CTLA-4,LAG-3in the CD8+T cells of the HBV infection patients with different clinicalmanifestations as well as the relationship between these factors and serology,virology,biochemical parameters respectively, the study aims at investigating the role ofHBsAg, HBeAg, the multiple inhibitory receptors and transcription factor T-bet in thepathogenesis of chronic hepatitis B.Method:1.99patients of HBV infection were enrolled, including20of acute hepatitisB(AHB),19of chronic hepatitis B virus carriers(AsC),25of chronic hepatitis B(CHB),23of inactive HBsAg carriers,12of chronic hepatitis B recovery,14normal people ascontrol. ALT, AST, HBV DNA, HBsAg, HbeAg, ultrasonic examination concerning liverand any other clinical data were examined;2. The peripheral blood mononuclearcells(PBMC) were separated, then the PBMC were collected and frozen;3. Thespecimens are divided into two groups: the one was non-stimulated group; the other onewith the stimulation of anti-CD28and HBV core protein antigen peptide. Flowcytometry were used to detect expression of INF-γ，PD-1, LAG-3, CTLA-4and T-bet,IFN-γ in the CD8+T cells;4. To analyze the differences and relationship of IFN-γ, T-bet, the inhibitory receptors of HBV infection patients with different clinical manifestationsas well as the relationship between these factors mentioned above and serology,virology,biochemical parameters.Results:1. The expression analyzes of the factors of the non-stimulated group：1) PD-1: AHB is the highest in all groups, which is higher than AsC, the inactive carrierand control. Control is lower than CHB and hepatitis recovery.The differences arestatistically significant.2) T-bet: AHB is the highest in all groups, which is higher than AsC, CHB, inactivecarriers and control. AsC is higher than control. The differences are statisticallysignificant.3) IFN-γ：AHB is the highest in all groups, and the difference is no statistic significance.2. The expression analyzes of the factors of the stimulated group.T-bet: CHB is the highest in all groups. AHB,CHB, hepatitis recovery are higher thancontrol，and the differences present statistic significance.3. The expression analyzes of the factors between the non-stimulated group and thestimulated group.1) AHB: T-bet of non-stimulated group is higher than the stimulated group, p=0.001.2) AsC: PD-1of non-stimulated group is lower than the stimulated group, p=0.0002.3) CHB: PD-1,IFN-γ and T-bet of non-stimulated group are lower than the stimulatedgroup. P values are0.004,0.033and0.045respectively.4) Inactive carriers: PD-1and IFN-γ of non-stimulated group are lower than thestimulated group. P values are0.001,0.009respectively.5) Hepatitis recovery: CTLA-4, PD-1and IFN-γ of non-stimulated group arelower than the stimulated group.P values are0.039,0.002,0.031respectively.6) Control: PD-1, IFN-γ and T-bet of non-stimulated group are lower than the stimulatedgroup. P values are0.0007,0.013and0.0007respectively.4. Correlation analysis of each group factors. 1) AHB: T-bet expression positively correlated with PD-1in stimulated group.2) CHB: T-bet expression positively correlated with CTLA-4, IFN-γ in non-stimulatedgroup and stimulated group.3) Inactive carriers: T-bet expression positively correlated with CTLA-4, LAG-3innon-stimulated group and stimulated group.4) Control:T-bet expression positively correlated with CTLA-4, PD-1, LAG-3innon-stimulated group and stimulated group.5. Correlation analysis between clinical indicators and each factors.1) AHB: HBeAg positively correlated with T-bet, and the difference is statisticallysignificant.2) AsC: HBsAg positively correlated with PD-1, IFN-γ,and the difference is statisticallysignificant.3) CHB: HBsAg positively correlated with CTLA-4, PD-1, IFN-γ, T–bet, and thedifference is statistically significant.Conclusion:These results reveal that AHB is higher than CHB concerning the expression level ofIFN-γ、T-bet and inhibitory receptors such as PD-1in CD8+T cells. It suggests that theCD8+T cells response in AHB is stronger than in CHB. In the non-stimulated group andstimulated group, T-bet expression positively correlated with the multiple inhibitoryreceptors, which are not accordance with the early literature about the mice infected bylymphocytic choriomeningitis virus. It shows that the inhibitory receptors maybe increasein a passive state to prevent the excessive immune damage. In addition, the inhibitoryreceptors play an important role during the HBV infection chronicity. HBsAg, HBeAgpositively correlated with T-bet，inhibitory receptors,suggesting that HBsAg and HBeAgmaybe have no inhibitory effect on the expression of T-bet. It needs study further whetherit could enhance expression of inhibitory receptors or not.. |
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