| Objective:Analysis changes of Th17/Treg balance and level of specific transcription factor(RORγt、Foxp3) in chronic hepatitia B patients, discuss the causes of Th17/ Treg imbalances resulting to the immunology mechanism of persistent infection of HBV and chronicity. Provide new idea for treatment of CHB patients, further to prevent cirrhosis and liver cancer associated hepatitis B.Methods:This study selected for 61 cases chronic hepatitia B patients who came to hospitalization or out-patient treatment in the Infectious Disease Department of The First Affiliated Hospital of Kunming Medical University from October 2013 to July 2014. Among them,41 cases were mild chronic hepatitis B patients,20 cases were moderate to severe hepatitis B patients. Select 20 cases who were normal physical examination people in our hospital from the same time period as control group at the same time. HBV-makers (includes:HBsAg,HBsAb,HBeAg,HBeAb,HBcAb) was detected by the ELISA way;The Liver function indicators such as ALT, AST, TBIL, were analyzed by the automatic biochemical analyzer. The HBV-DNA levels of the patients group were tested by real-time fluorescent quantitative PCR analyzer. The frequency of Thl7cells and Treg cells in the peripheral blood of the patients group and control group were detected by flow cytometry. Peripheral blood transcription factor (retinoid-related orphan nuclear receptor yt, RORyt) and (forkhead family transcription factor 3, Foxp3) were detected by real time quantitative RT-PCR.Statistic analyzsis of All date was conducted by the SPSS17.0 statistical software.Results:1. Th17/CD4+T cells ratio and RORyt mRNA level of CHB patients peripheral blood were higher than the control group, the difference was statistically significant; Th17/ CD4+T cells ratio and RORyt mRNA level of CHB patients peripheral blood of moderate to severe group was higher than the mild group and control group, the difference was statistically significant, but when the mild group compared with the control group, Th17/CD4+Tcells ratio and RORyt mRNA level of CHB patients peripheral blood had not obvious difference.2. Treg/CD4+T cells ratio and Foxp3mRNA level of CHB patients peripheral blood were higher than the control group, the difference was statistically significant; Treg/ CD4+T cells ratio and Foxp3mRNA level of CHB patients peripheral blood of moderate to severe group was higher than the mild group and control group, the difference was statistically significant. Treg/CD4+T cells ratio of CHB patients peripheral blood of the mild group was higher than control group.But when the mild group compared with control group, Foxp3mRNA level of CHB patients peripheral blood had not obvious difference.3. Th17/Treg cells ratio in CHB patients is higher than the control group, the difference was statistically significant; Th17/Treg ratio of the moderate to severe group was higher than the mild group, the difference was statistically significant, but the moderate to severe group and mild group compared with control group, no difference of Th17/Treg cells ratio. Th17/CD4+T cells ratio, Treg/CD4+T cells ratio and Th17/Treg cells ratio between HBeAg (+) group and HBeAg (-) group have no obvious difference.4. Th17/CD4+T cells ratio of the mild and the moderate to severe group were positively related to the level of ALT and RORyt mRNA, the difference was statistically significant, but with have no obvious correlation between the AST, TBIL and HBV-DNA level; Treg/CD4+T cells ratio of the mild and the moderate to severe group and Foxp3mRNA levels were positively correlated, the difference was statistically significant, but no obvious correlation between ALT, AST, TBIL and HBV-DNA level. RORyt mRNA and Foxp3mRNA of the mild and the moderate to severe group respectively with ALT levels were positively correlated, the difference was statistically significant. Th17/CD4+T cells ratio was positively related to Treg/CD4+T cells ratio in CHB patients.5. Th17/Treg cells ratioof HBeAg (+) group was positively correlated with ALT, AST levels, the difference was statistically significant, but no obvious correlation between the TBIL and HBV-DNA level; Thl7/Treg cells ratio of HBeAg (-) group were positively correlated with TBILiL level, the difference was statistically significant, but no obvious correlation between ALT, AST and HBVDNA level; Th17/Treg cells ratio of the mild and the moderate to severe group had no obvious correlation between ALT, AST, TBIL, HBV DNA levels and RORytmRNA/Foxp3mRNA ratio.Conclusion:Frequency of Treg and Th17 cells and transcription factors RORyt and Foxp3 level in peripheral blood of CHB patients were higher than the control group. However Treg cells increase was less than Th17 cells increase, resulting to Th17/ Treg balance is broken, immune pattern of Th17/Treg moved to Th17 cells. Increased Th17 cells may be one of main reasons of causing the liver inflammation, liver cell damaging, and is closely related to severity of CHB. Treg cells high expression in CHB patients can down regulate cellular immune function ability of removing HBV that may become one of the causes of HBV chronicity and persistent infection. Th17 cells in chronic hepatitis B patients can avoid the Treg cells to protect HBV when the virus persists in the body, so Th17/Treg balance is important to the occurrence, development and outcome of CHB. At the same time, we think about that the level of Th17 cells and Treg cells has no influence on HBVDNA replication. |
PDF Full Text Request