Implication Of Th17 And Th1 Cells In Patients With Chronic Active Hepatitis B And Acute-On-Chronic Hepatitis B Virus Liver Failure

第一部分IMPLICATION OF TH17 AND TH1 CELLS IN PATIENTS WITH CHRONIC ACTIVE HEPATITIS BBackground Hepatitis B virus (HBV) infection is a major public health threat in the world, especially in China. Globally, there are approximately 350 million carriers of HBV, of whom 0.5-1.0 million die due to the HBV-associated liver disease each year. Both CD4+(Th) and CD8+(Tc) T lymphocytes can be functionally divided into type 1(T1) and type 2(T2) subsets based on the secretion of either IFN-γ(T1) or IL-4 (T2). Recent studies have provided compelling evidence for a third CD4+ T cell effector subset besides the well-described Th1 and Th2 CD4+ T cell. The newly identified CD4+ T effector cells have been named as Th17 cells. These cells are characterized as preferential producers of IL-17A (also known as IL-17), IL-17F, IL-21, IL-22, and IL-26 in humans. Retinoid orphan nuclear receptor (RORC), which encodes the human ortholog of mouse RORct, is a key regulator of Th17-cell lineage differentiation. Th17 cells and their effector cytokines are increasingly being recognized as key determinant in the induction of autoimmune diseases and malignant tumor as well as alcoholic liver disease. There are limited data to date about Th17 levels in patients with chronic hepatitis B (CHB).To further investigate the role of Th17 and Thl cells, as well as their relationship in the pathogenesis of CHB, we examined the frequency of Th17 and Thl cells in CHB active patients through intracellular cytokines analysis by flow cytometry, ELISA and real-time PCR.Objective The pathogenesis of hepatitis B virus (HBV) associated chronic liver disease is still not fully understood.The immune imbalance of cytokine profile exerts a profound influence on the resolution of HBV infections and HBV clearance. This present study aimed to evaluate the immune status of the peripheral T helper (Th) 17 and Th1 cells in the active patients with chronic HBV infection. Methods Thirty patients with chronic active hepatitis B were included in our present study. The frequency of peripheral Th 17 cells (CD3+CD8-IL-17+T cells), Thl cells (CD3+CD8-IFN-γ+T cells), and Tc1 cells (CD3+CD8+IFN-γ+T cells) in chronic hepatitis B(CHB) were analyzed by flow cytometry. The protein and mRNA levels of interleukin-17 (IL-17) and interferongamma (IFN-γ) were measured by enzyme-linked immunosorbent assay and quantitative real-time polymerase chain reaction (PCR).Results The percentage of Th17 cells in peripheral blood of CHB patients (1.53±0.52%) was significantly increased than that in normal controls (0.92±0.20%; P<0.05). In contrast, the percentage of Th1 and Tc1 cells of CHB patients was significantly decreased as compared with that of control group. The frequency of Th17 cells had a negative correlation with Th1 cells, and a positive correlation with serum alanine aminotransferase in CHB patients.Conclusion Our study data strongly support a high Th1 and Tc1 polarization of the immune response in CHB and CTL-mediated cytotoxicity is an alternative mechanism in CHB. More important, our study demonstrates for the first time that Th17 may be an important determinant in the evolution of CHB along with Th1 and Tc1, suggesting that blocking the abnormality of Th17 cell in CHB is likely a promising therapeutic concept for CHB. 第二部分IMPLICATION OF TH17 AND TH1 CELLS IN PATIENTS WITH ACUTE-ON-CHRONIC HEPATITIS B VIRUS LIVER FAILUREBackground Hepatitis B virus (HBV) infection still poses a major public health threat in a large part of the world. Some CHB patients may rapidly progress towards liver failure, a condition referred to as acute-on-chronic liver failure(ACLF). In China, acute-on-chronic hepatitis B liver failure (ACHBLF) accounts for more than 80% of ACLF cases due to a high incidence of HBV infection. ACLF shares striking similarities with septic shock with regard to the features of systemic inflammation, progression to multiple organ dysfunction and functional immunoparesis.The precise mechanisms underlying the deterioration of liver function occurring in ACLF remain unclear.There are limited data to date about Th17 levels in patients with acute-on-chronic hepatitis B liver failure (ACHBLF).To further investigate the role of Th17 and Thl cells, as well as their relationship in the pathogenesis of ACHBLF, we examined the frequency of Th17 and Th1 cells in ACHBLF active patients through intracellular cytokines analysis by flow cytometry and ELISA.Objective Acute-on-chronic hepatitis B virus liver failure (ACHBLF) has been shown to carry poor prognosis, however, the pathogenesis of ACHBLF is still not fully understood. This present study aimed to evaluate the immune status of the peripheral T helper (Th) 17 and Thl cells in the active patients with ACHBLF.Methods Twenty patients with ACHBLF were included in our present study. We examined the levels of Th17,Th1 and Tc1 cells in peripheral blood which was activated in vitro by PMA/ionomycin in short-term cultures in ACHBLF patients and controls by flow cytometry through intracellular cytokines analysis. Th17 cells and Thl cells were identified as those that were CD3+CD8-IL-17A+and CD3+CD8-IFN-γ+, and Tc1 cells were those that were CD3+CD8+IFN-γ+. ELISA was used to detect the expression of Th1 and Th17 cytokines (IFN-γand IL-17) in plasma. Results The results showed that the percentages of both Thl and Tc1 cells in ACHBLF patients increased significantly compared with the normal controls (P<0.01 for Thl, P<0.05 for Tc1). More importantly, the percentage of Th17 in patients with ACHBLF was markedly higher than that of normal controls (P＜0.05). Also, the percentage of Th17 cells positively correlated with Thl cells (r=0.61, P＜0.05). There was no significant correlation between Th17 and Tc1 cells (r=0.09, P=0.71).Conclusion Our study demonstrates for the first time that Th17 cells may be an important determinant in the evolution of ACHBLF along with Thl cells, suggesting that blocking the abnormality of Th17 cell in ACHBLF is likely a promising therapeutic concept for ACHBLF.