Correlation between programmed death-1 and T-cell immunoglobulin mucin-3 co-expression on hepatitis B virus-specific CD8+ T cells and liver inflammatory activities in patients with chronic hepatitis B virus infection / 中华传染病杂志

ABSTRACT

Objective To study programmed death-1 (PD-1) and T-cell immunoglobulin mucin3 (Tim-3) co-expression on peripheral blood mononuclear cells (PBMC) and hepatitis B virus (HBV)-specific CD8+ T cells in patients with chronic HBV infection and its correlation with liver inflammatory activities.Methods One hundred and sixty subjects with chronic HBV infection who visited the outpatient and inpatient Department of Infectious Diseases in the Third Affiliated Hospital of Sun Yatsen University were enrolled,including 63 cases of mild or moderate CHB (MCHB),31 of severe CHB (SCHB),55 of acute-on-chronic liver failure (ACLF) and 11 inactive HBsAg carriers.Twenty healthy subjects were enrolled as controls.Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect HBV DNA,enzyme-linked immunosorbent assay (ELISA) to measure HBV serological markers,and flow cytometry to detect human leukocyte antigen (HLA)-A2 and determine the expression of PD-1 and Tim-3 on PBMC and HBV-specific CD8+ T cells.Cell counts of Tim-3+,PD-1+,and Tim-3+/PD1+ PBMC and HBV-specific CD8+ T cells were compared among different groups,and their correlation with commonly used inflammatory activity indicators were studied.Analysis of variance and Kruskal-Wallis test were used for measurement data with gaussian distribution and skewed distribution,respectively.Spearman correlation analysis was used to assess the association between Tim-3/PD-1 expression and inflammatory activity indicators.Results The frequency of Tim-3+/PD-1+ PBMC was 0.25 ％ in ACLF group (P=0.0049),0.24 ％ in SCHB group (P-0.0025) and 0.13％ in MCHB group (P=0.0006),which were significantly higher than that in control group (0.03％),and the frequency of Tim-3 /PD-1-PBMC in the three groups were significantly lower than that in control group (P =0.0000),but the differences between HBsAg carriers (0.10％) and controls (0.03％) were not statistically significant (P=0.28).Among 160 subjects,78 were HLA-A2 positive.The frequency of Tim-3+/PD-1+ HBV-specific CD8+ T cells was 68.72％±17.21％ in ACLF group,and 59.66％± 19.25％ in SCHB group,which were significantly higher than that in HBsAg carrier group (33.93％ ± 10.80％,P=0.0000,P=0.0054).The frequency of Tim-3 /PD-1-HBV-specific CD8+ T cells in ACLF group was 2.80％,which was significantly lower than that in HBsAg carrier group (27.24％,P=0.0004).The frequency of Tim3+/PD-1+ HBV-specific CD8+ T cells was positively correlated with alanine aminotransferase (r=0.26,P=0.022),aspartate aminotransferase (r=0.28,P=0.012) and total bilirubin levels (r=0.36,P=0.001); and inversely correlated with albumin level (r=-0.35,P=0.002) in serum.Furthermore,it was positively correlated with international normalized ratio (INR,r=0.34,P =0.045) and model for end-stage liver disease score (r=0.43,P=0.027) in ACLF group.Conclusions Co-expressions of Tim-3 and PD-1 on PBMC and HBV-specific CD8+ T cells are significantly upregulated in patients with CHB,and correlated with liver inflammatory activities.These findings indicate that Tim-3 and PD-1 co-expression may involve in disease activity and liver failure in CHB.