Detection And Clinical Significance Of The Unbalance Of Tfh And Treg Cells And Interleukin-21 In The Peripheral Blood Of Patients With Chronic HBV Infection

Objective: To investigate the role of the unbalance of Tfh and Treg cells and interleukin-21 in the occurrence, development and outcome of chronic HBV infection,by detecting the populations of Tfh and Treg cells and interleukin-21 in the peripheral blood of patients with chronic HBV infection.Methods:1. 20 healthy donors(HC), 14 patients with Hepatitis B virus carrier(ASC), 50 patients with Chronic hepatitis B(CHB), 20 patients with HBV-related hepatocirrhosis and 24 patients with HBV- related Acute On Chronic Liver Failure were selected. The frequency of Tfh cells and Treg cells in the peripheral blood were detected by flow cytometry. Compare the frequency of Tfh, Treg cells and the ratio of Treg/Th17 among five groups. Analysis the above indicators correlated with clinical status, such as HBV-DNA, PT, MELDand biochemical indicators(ALT, AST, TBil, DBil).2. The cytokines in peripheral plasma IL-21 was measured by enzyme-linked immunosorbent assay(ELISA). Analysis the differences of IL-21 among five groups,and the correlation with HBV-DNA, PT, MELDand biochemical indicators(ALT, AST,TBil, DBil).Result:1. The frequency of Tfh cells in the peripheral boold of in the HC group, ASC group, CHB group, HBC-LC group and ACLF group were increased(χ2=45.827,P<0.001). There was significant difference betweem any two of groups among five groups, while there was no significant difference between HC group and ASC group(Z=0.774, P=0.439). The frequency of Treg cells were increased in each group(χ2=5.747, P=0.219). Two groups intercomparison among five groups, only- 6-frequency in the CHB group and ACLF group were higher in the HC group(Z=-2.122,P=0.034; Z=-2.241, P=0.025). There were not statistically difference among five groups of the ratio of Tfh/Treg, while there was significant difference with HC group and HBV-LC group(Z=-2.079,P=0.039). The frequency of Tfh cells were positively correlated TBil, DBil,AST, ALB and the ratio of Tfh/Treg(P<0.01). The frequency of Treg cells were positively correlated biochemical function(TBil, DBil and AST)(P<0.05), and which was a negative relation between Treg cells and the ratio of Tfh/Treg(P<0.01), while there was no significant correlation between the ratio of Tfh/Treg and the above indicators(P>0.05).2. The result of subgroup analysis: Tfh cells frequency were significantly lower in the HBe Ag-positive CHB patients than in HBe Ag-negative CHB patients(P<0.05),while there was no significant difference between two groups of the frequency of Treg cells and the ratio of Tfh/Treg. The three indicators showed no significant statistical significance between all groups of complications of HBV- LC group. The Tfh cells frequency were higher in survival group and non-survival group than in the HC group(P<0.01), but there was no significant difference with in survival group and non-survival group(P>0.05). The frequency of Treg cells in the non-survival group,survival group and HC group were decreased(P<0.05), but there was no significant difference with in survival group and non-survival group(P>0.05). The ratio of Tfh/Treg in the non-survival group, survival group and HC group were decreased(P>0.05).3.There was no significant difference in levels of IL-21 between the five groups,while there was significant difference with HC group and ACLF group(P<0.05). In the comparism of subgroup, the level of IL-21 was showed no significant statistical significance among all groups(P>0.05), while there was no significant difference between other group. There was no significant correlation between Tfh cell, Treg cell,the ratio of Tfh/Treg and the clinical status(P>0.05).4. After two weeks of comprehensive treatment, the above indicators were compared between treatment before and treatment after in ACLF group: The Tfh cells frequency and the ratio of Tfh/Treg were significantly declined in the group oftreatment after than in the group of treatment before(P<0.01), but the frequency of Treg cells was slight increased(P>0.05), and the level of IL-21 was declined(P>0.05).Conclusion: The ratio of Tfh/Treg were showed different degree of imbalance in the every phase of chronic HBV infection, and with the progress of the disease, the unbalance degree was more obviously, which indicated that it may play a potential role in the occurrence, development and outcome of chronic HBV infection. And the frequency of Tfh cells in the peripheral boold played a leading role in the Tfh/Treg imbalances. The Tfh cells and IL-21 involved in pathogenic process of each stage of chronic HBV infection.