2A) There were no significant differences observed in the freque

2A). There were no significant differences observed in the frequencies of IL-22–producing CD4+ T cells and IL-22–producing Th17 cells among these three groups of subjects (Supporting Fig. 2B,C). Antigen-specific Th17 cells have been described in HCV infection,23 but it is unknown whether HBV-specific Th17 cells will be present in

patients with CHB. Our data indicated that PBMCs from patients with CHB expressed high levels of RORγt and IL-17 mRNA in response to HBcAg (Fig. 2D). Simultaneously, these PBMCs could also produce median amounts of IL-17A after check details HBcAg stimulation (Fig. 2E). These capacities of PBMCs to express RORγt and IL-17 mRNA and produce IL-17A in response to HBcAg were largely reduced selleck compound after deletion of CD4+ T cells from PBMCs in patients with CHB (Fig. 2D,E). These data clearly indicated that in CHB patients there are some HBV-specific Th17 cells

displaying responsiveness to HBcAg. We analyzed the correlation between Th17 frequency and plasma HBV DNA load or serum alanine aminotransferase (ALT) levels in these CHB and ACLF patients. There were some significant positive correlations between Th17 frequency and both plasma HBV DNA load (r = 0.212, P = 0.024; Fig. 3A) and serum ALT levels (r = 0.390, P < 0.001; Fig. 3B) in these HBV-infected subjects. Further analysis indicated that these positive associations occurred only in patients with CHB (Fig. 3A,B) but not in patients with ACLF. In addition, we also found that CHB patients with high HAI scores (G2-G3) (n = 12) had a greater proportion of Th17 cells than did CHB patients with low HAI scores (G0-G1) (n = 9) (Fig. 3C). These data suggest that peripheral Th17 cell frequency is closely associated with liver injury, indicated by serum ALT levels and liver HAI scores in CHB patients. We also examined the distribution of IL-17+ cells selleck chemicals in the livers of CHB patients. As shown

in Fig. 4A, tonsil tissue from a healthy individual, which served as a positive control, showed obvious IL-17 staining, whereas the liver tissue from a healthy donor had few IL-17+ cells. Interestingly, more IL-17+ cells were found accumulated in the lobular and portal areas of livers in CHB patients (Fig. 4B). The liver-infiltrating IL-17+ cells were differentially distributed in CHB patients with varying G scores: more IL-17+ cells were found to be infiltrated in the livers of patients with a G4 score than those of patients with G2 and G1 scores (Fig. 4B). Using double immunostaining we confirmed that intrahepatic IL-17+ cells were primarily expressed on CD4+ T cells (Fig. 4C). Quantitative analysis of intrahepatic IL-17+ cells documented that livers from CHB patients exhibited more IL-17+ cell infiltration than did livers from HC subjects. In addition, in the lobular area of patients with a G4 score the number of IL-17+ cells per hpf was significantly more than in patients with G2-G3 scores and in HC subjects (Fig. 4D; all P < 0.01).

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