| Objective:This study was to investigate the role of imbalance between Th17 cells and Treg cells in severe acute exacerbation of chronic hepatitis B, and to provide the experimental basis for the judgement of chronic severe hepatitis B progression. Methods:According the criteria: time of disease onset< 2 weeks, ALT>800U/L, TBIL<171 ummol/L, PTA>40%, and no complications during hospitalization, fifty- two patients with hepatitis B were included in our study. By observing whether patients progressed to chronic severe hepatitis B during hospitalization, we divided those patients into two groups: clinical exacerbation(19 cases) and improvement(33 cases). Peripheral blood and serum was collected at baseline, 1 or 2 weeks post-treatment and at the end of the treatment. The frequencies of Th17 and Treg cells were analyzed by flow cytometry, and the Th17/Treg ratio was subseqently calculated. Enzyme- linked Immunosorbent Assay(ELISA) was used to determine the levels of IL-17, IL-10 and TGF-β in the serum. The frequencies of Th17 and Treg cells, Th17/Treg ratio and cytokine levels of each group at indicated timepoints were compared, then the relevance between the Th17/Treg ratio and hepatitis clinical parameters and cytokine was analyzed. Results:1) At baseline, 1 and 2 weeks after treatment, and treatment endpoint, the frequencies(%) of Th17 cells in exacerbation group were 1.71±0.67, 2.36±0.77, 3.15±0.93 and 1.54±0.89 respectively. We observed that the Th17 frequency increased gradually, peaked at 2 weeks post-treatment, then decreased at the treatment endpoint, and the comparisons between each of them were stat istically significant(P <0.05).While the frequencies(%) of Th17 in improvement group were 1.60±0.67, 1.14±0.52, 0.86±0.43 and 0.58±0.37 respectively. We observed that the Th17 cells frequencies(%) decreased gradually and stayed at a relatively low level. In addition, Th17 frequencies in exacerbation group were higher than those in improvement group at every time point,except baseline(P < 0.05 respectively).2) At baseline, treatment 1 and 2 weeks after treatment, and treatment endpoint, the frequencies(%) of Treg cells in improvement group were 4.95±2.17, 7.32±2.61, 6.76±2.33 and 5.13±2.31 respectively, and the comparisons between baseline vs 1 week post-treatment and 2 week post-treatment vs treatment endpoint were statistically significant(P<0.05). We observed that the Treg frequency increased gradually, peaked at 1 week post-treatment, then decreased slightly.While the frequencies(%) of Treg cells in exacerbation group were 3.28±1.28, 2.53±1.01, 2.18±0.79 and 1.38±0.52 respectively, and the comparison between 2 weeks post-treatment vs treatment endpoint was statistically significant(P <0.05).. We also observed the Treg frequency decreased gradually and stayed at a relatively low level.Morever, the Treg frequencies in improvement group were higher than those in exacerbation group at every time point(P < 0.05 respectively).3) At baseline, treatment 1 and 2 weeks after treatment and treatment endpoint, the ratio of Th17/Treg in exacerbation group were 0.56±0.23, 0.98±0.28,1.61±0.55 and 1.17±0.64 respectively, and the comparisons between each of them were statistically significant(P<0.05).We observed that the ratio of Th17/Treg increased gradually and stayed at a relatively high level with the development of the disease, peaked at 2 weeks post-treatment, then decreased at the treatment endpoint.While the ratio of Th17/Treg in improvement group were 0.34±0.11, 0.18±0.07, 0.11±0.03 and 0.12±0.56 respectively, and the comparisons between baseline vs 1 week post-treatment and 1 week post-treatment vs 2 weeks post-treatment were statistically significant(P<0.05). We also observed that the ratio of Th17/Treg decreased gradually and stayed at a relatively low level. In addition, the ratio of Th17/Treg in exacerbation group were higher than those in improvement group at every time point(P <0.05 respectively).4) With disease progression, IL-17 levels increased gradually and stayed at a relatively high level while the levels of IL-10 and TGF-β decreased gradually and stayed at a relatively low level in exacerbation group. In contrast, we observed opposite results in improvement group. In addition, the levels of cytokines in exacerbation group were higher than those in improvement group at every time point(P <0.05 respectively).5) At baseline, the levels of ALT, AST, TBil, DBil and PT in exacerbation group were significantly higher than those in improvement group, and the levels of ALB and PTA in exacerbation group were lower than those in improvement group(P<0.05 respectively).6) Both in exacerbation group and improvement group, Th17/Treg ratio was positively correlated with serum total bilirubin, but negatively correlated with prothrombin activity. In addition, Th17 positively correlated with IL-17, and Treg positively correlated with IL-10 and TGF-β.Conclusions:The results of 52 patients with chronic hepatitis B in this study showed that, patients in exacerbation group had higher levels of Th17 cells and its related factors than those in improvement group. We also found that patients in improvement group had higher levels of Treg cells and its related factors than those in exacerbation group, suggesting that the trend of the changes of frequency of Th17 cells and Treg cells was opposite. This situation illustrates that Th17 cells can promote inflammation, which facilitates the progression of hepatitis B infection to gravis hepatitis B. We also believe that Treg cells can control the degree of inflammation, avoid excessive liver cell damage and promote disease remission. And Th17/Treg ratio maybe as a immune biological marker judgging the severity in the process of the disease. |
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