Detection And Clinical Significance Of Treg And Th17Cells And Treg/Th17Cell-associated Cytokines In The Peripheral Blood Of Patients With Chronic Hepatitis B

Objective: To investigate the role of Treg cells and Th17cells and Treg/Th17differentiation-related cytokines in the occurrence, development and outcome ofchronic hepatitis B (CHB), by detecting the populations of Treg and Th17cells andtheir associated cytokines in the peripheral blood of CHB patients.Methods:1. Relationship between Treg cells, Th17cells and the clinical indicators.46patients with low or moderate Chronic hepatitis B (CHB-LM),24patients with severechronic hepatitis B (CHB-S) and20healthy donors (HC) were selected. Thefrequency of Treg cells and Th17cells in the peripheral blood were detected by flowcytometry. Compare the frequency of Treg, Th17cells and the ratio of Treg/Th17between HC, CHB-LM and CHB-S group. Analysis the above indicators correlatedwith clinical status, such as gender, age, HBeAg status, disease severity andbiochemical indicators (ALT, AST, TBil, DBil).2. Correlations between Treg/Th17cell-associated cytokines in plasma andbiochemical indicators. The related cytokines in peripheral plasma IL-10, TGF-β1,IL-17and IL-23were measured by enzyme-linked immunosorbent assay (ELISA).Analysis the differences of cytokine concentration among HC group, CHB-LM groupand CHB-S group, and the correlation with ALT, AST, TBil and DBil.3. Compare the frequency of Treg, Th17cells, the ratio of Treg/Th17and theirrelated cytokines in the peripheral blood of patients with CHB before and afterantiviral therapy, and whether it remains a correlation with ALT, AST, TBil and DBil. Result:1. The frequency of Treg cells in the peripheral boold of in the HC group,CHB-LM group and CHB-S group were increased (5.83±1.07%,6.78±2.53%,7.55±2.11%, χ2＝7.914, P=0.019). The frequency of Th17cells were significantlyincreased in each group (χ2=10.41, P=0.005). Th17cells frequency were significantlyhigher in the CHB-LM group (0.22±0.14%, P<0.01) and CHB-S group0.24±0.20%,P<0.01) than in the HC group (0.11±0.05%), but there was no significant differencewith CHB-LM group and CHB-S group (P>0.05). Compare with HC group, thefrequency of Treg cells and Th17cells were significantly increased in CHB group(7.04±2.41%vs5.83±1.07%, P=0.025;0.23±0.16%vs0.11±0.05%, P=0.005), whilethe ratio of Treg and Th17was lower than HC group (56.72±60.37vs76.09±68.49,P=0.047).The ratio of Treg/Th17was significantly lower in the CHB-LM group(49.13±57.73) than in the the CHB-S group (68.79±61.23, P<0.05), and there were nodifference in different gender, age and HBeAg status (P>0.05). The frequency of Tregcells and Th17cells were no difference in different gender, age, severity of diseaseand HBeAg status (P>0.05). The frequency of Treg cells were positively correlatedbiochemical function (TBil, DBil and AST)(P<0.05) in CHB patients, while therewas no significant correlation between Th17cells and the above indicators (P>0.05).The ratio of Treg/Th17was positively correlated with TBil, DBil and ALT inHBeAg-positive CHB patients (P<0.05), while it were negatively correlated with ALTin HBeAg-negative CHB patients (P<0.05).2. The levels of IL-10and IL-23were significantly higher in CHB-LM andCHB-S group than in the HC group (P<0.05), while there was no significantdifference in the CHB-LM group and CHB-S group. The levels of TGF-β1weresignificantly higher in the CHB-S group than in the CHB-LM group and HC group,while there was no significant difference in the CHB-LM group and HC group. Theplasma IL-17showed an increasing trend in the three groups (χ2=0.85, p=0.645), therewas no difference between each group. IL-10levels were positive correlated withIL-23(r=0.902, P<0.01). TGF-β1was positively correlated with TBil and DBil.3. The frequency of Treg cells and their related cytokines IL-10, TGF-β1and IL-23were significantly decreased in CHB patients (P<0.05), while the frequency ofTh17cells and IL-17were increased, and the ratio of Treg/Th17were decreased(P>0.05). Treg cells were positively correlated with Treg/Th17ratio and TBil, DBiland AST. Th17cells were negatively correlated with Treg/Th17ratio, while withoutcorrelation with TBil, DBil, ALT and AST. IL-10was positively correlated with IL-23(r=0.965, P<0.01). IL-17was positively correlated with biochemical indicators (ALT,AST). Further analysis of the follow-up data of HBeAg-positive CHB patients, wefound Treg/Th17ratio were positively correlated with TBil, and it showed a positivetrend with other biochemical indicators (ALT).Conclusion: The ratio of Treg/Th17was decreased in CHB patients. With thedegree of inflammation increased, the ratio of Treg/Th17was significantly increased.After effectively treatment, Treg/Th17ratio continue to decrease. Thses indicated thatTreg/Th17dysfunction exists in patients with CHB, and it may play a potential role inthe onset and development of CHB. Th17cells and IL-17were nothing to do with thegrade of liver injury. Treg cells and Treg/Th17ratio are related with the grade of liverinjury, TGF-β1may play an important role in immunoregulatory.For HBeAg-positiveCHB patients, Treg/Th17ratio may be a valid immune indicator to assessment theefficacy of treatment.