Effects Of Antiviral Therapy On Non-specific T-cell Proliferation And HBV-specific T-cell Responses In Chronic Hepatitis B Patients

Objective:（1）Detect the non-specific T-cell proliferation and HBV-specifc T-cellresponses in chronic hepatitis B patients without antiviral treatment. To assessthe characteristics of T cell immune function in CHB patients.（2）Detect the non-specific T-cell proliferation and HBV-specifc T-cellresponses in CHB patients with antiviral treatment. To investigate the effectsof antiviral therapy on T cell immune response.（3）analyze the relationship between non-specific T-cell proliferation,HBV-specifc T-cell responses and liver function indicators, HBV makers. Toexplore the relationship between T cell immune response and HBV viralkinetic features.Methods:（1）17CHB patients without antiviral treatment (untreated group),20CHB patients with antiviral treatment (treatment group), and10healthy adultsinoculated hepatitis B vaccination successfully (healthy group) were enrolledthis study. The peripheral blood monocular cells (PBMCs) of CHB patientswere separated from the whole blood. Then assess the non-specific T-cellproliferation and analyze the effects of antiviral therapy.（2）Another83patients with HBV infection were enrolled this study,included23CHB patients without antiviral treatment (untreated group),47CHB patients with antiviral treatment (treatment group), and15acute hepatitis B patients (AHB group). Then separate PBMCs from CHB patients,assess the HBV-specific T-cell responses, and analyze the effects of antiviraltherapy to HBV-specific T-cell responses.（3）The non-specific T-cell proliferative response was evaluated byCFSE Cell Proliferation Kit and flow cytometry in the presence anti-CD3anti-CD28mixture. And the HBV-specific T-cell responses to HBsAg andHBcAg were measured by ELISPOT assay detecting interferon-gamma in thepresence of HBsAg overlapping peptides (S pool) and HBcAg overlappingpeptides (C pool).（4）Compare the differences of non-specific T-cell proliferation andHBV-specifc T-cell responses between untreated group and treatment group,analyze the effects of antiviral therapy on non-specific/HBV-specifc T cellimmune response, and explore the relationship between T cell immuneresponse and HBV viral kinetic features.Results:（1）The non-specific proliferation response of PBMC from untreatedgroup was weak. And the percentage of proliferative cell in untreated groupwas significant lower than treatment group (t=4.229, P<0.001) and healthygroup(t=6.800, P<0.001), respectively. In addition, The non-specificproliferation response of PBMC from treatment group was as vigorous ashealthy group (t=0.670, P=0.509). Furthermore, The non-specificproliferation response of CD4+/CD8+T cell in untreated group wassignificant lower than treatment group (CD4+T cell: t=2.996, P=0.005;CD8+T cell: t=2.962, P=0.005) and healthy group (CD4+T cell: t=3.381,P=0.002; CD8+T cell: t=2.863, P=0.008), respectively. And the non-specificproliferation response of CD4and CD8double negative lymphocytes was no difference among the three groups (P>0.05).（2）The frequency and strength of HBV-specific T-cell responses toHBsAg and HBcAg were low and weak. However, CD28functional antibodydid amplify this response specifically. Differences were significant before andafter adding CD28functional antibody (Spool: t=-2.983, P=0.015；Cpool:t=-3.295, P=0.009).（3）Besides the low and weak HBV-specific T-cell responses, TheHBV-specific T-cell responses against HBsAg and HBcAg was no differencein untreated CHB patients(Z=-0.483, P=0.629). However, the CHB patientsafter antiviral treatment displayed a more robust HBV-specific T-cell responsethan untreated group (Spool: Z=-3.371, P=0.001; Cpool: Z=-2.510, P=0.012).In addition, the restoration of HBsAg-specific T-cell response was better thanHBcAg.（4）The non-specific proliferation response of CD4+/CD8+T cell inuntreated group were negatively correlated with serum HBV-DNA load(CD4+T cell: r=－0.536, P=0.027; CD8+T cell: r=－0.594, P<0.012). Thenon-specific proliferation response of CD4+T cell in untreated group werenegatively correlated with serum HBeAg level (r=－0.559, P=0.020).Moreover, HBV-specific T-cell responses against HBsAg and HBcAg werenegatively correlated with serum HBV-DNA load (Spool: r=－0.695, P<0.001;Cpool: r=－0.582, P=0.004). And the restoration of HBV-specific T-cellresponse was time dependent, which was increased with the duration ofHBV-DNA negative conversion, one year later recovered up to the peak, andthen declined with the time extension. Conclusion:According to our data, we have detected a weak non-specificproliferative response, a exhausted HBV-specific T-cell responses and anabnormal B7-CD28costimulatory signaling pathways in CHB patientswithout antiviral treatment. These functional defects may be associated withlong-term exposure to a large number of HBV antigen. However, antiviraltreatment can restore the non-specific T-cell proliferation and HBV-specifcT-cell responses significantly in CHB patients, while effectively suppressHBV-DNA synthesis and decline serum HBeAg level. Furthermore therestoration of HBV-specific T-cell response was time dependent, which wasrecovered up to the peak after HBV-DNA negative conversion one year, andthen declined with the time extension.In conclusion，This study confirmed HBV-specific T-cell exhaustion，discovered deficient non-specific proliferative response and abnormalB7-CD28costimulatory signaling pathways, clarified the effects of antiviraltherapy on T-cell immune responses, and illustrated the relationship betweenT cell immune response and HBV viral kinetic features in CHB patients.