Specific Function Of Peripheral Blood Lymphocytes With Chronic Hepatitis B Lymphocyte Function-specific During Anti-viral Therapy

Objective:To investigate the significance of specific function of peripheral blood lymphocytes with chronic hepatitis B lymphocyte function-specific during Nucleic acids anti-viral therapy.Methods:73 cases of patients(46 male and 27 female) with chronic hepatitis B,and 60 cases of asymptomatic carriers (42 male and 18 female ) were included in this study.53 patients with covert HBV infection (30 male and 23 female) and 30 healthy human (18 male and 12 female) were detected as control group .To selecte HLA-A2-positive individuals by flow cytometry using direct immunofluorescence method.ELISPOT assay was used to detect the frequency of secreting IFN-γcells of PBMC stimulated by HLA-A2-restricted CTLs epitopes of HBV (HBcAg18-27V and HBcAg18-27I) and to analyze the difference of antigen-epitope-specific CTL response in each group.The Hepatitis B viral infection marker of the patients who received antiviral therapy (Lamivudine and Adefovir dipivoxil were taken in turn) were detected by ELISA before and after the treatment. The liver function and the HBV-DNA were also detected at the same time. The frequency of secreting IFN-γcells of PBMC stimulated by HLA-A2-restricted CTLs epitopes of HBV (HBcAg18-27V and HBcAg18-27I) were detected by ELISPOT before the treatment and after 52 weeks'therapy.Results:1. In patients with chronic HBV infection,38 of the 73 patients with CHB (52.1%) were HLA-A2 positive and 15/30 (50.0%) in healthy human were HLA-A2 positive . With covert HBV infection, 31 of the 60 patients 31(51.7%) were HLA-A2 positive and 26/53 (49.1%) in asymptomatic carriers were HLA-A2 positive. There was no significant difference of the gene frequency distribution between them (P> 0.05).2. The ELISPOT results with chronic hepatitis B patients:⑴The rates of response to the two peptides were similar in each group.In the patients with covert HBV infection the rates of response to the two peptides were obviously higher than overt infection group (P <0.05).Both of these groups were obviously higher than asymptomatic carriers and control group (P<0.05). But there is no statistically significance between asymptomatic carriers and control group (P> 0.05).⑵The average response intensities to the two peptides were similar in each group. In the patients with covert HBV infection the average response intensities to the two peptides were obviously higher than overt infection group (P <0.05). Both of these groups were obviously higher than asymptomatic carriers and control group (P <0.05). But there is no statistically significance between asymptomatic carriers and control group (P> 0.05).3. The comparison between groups, the rates of response and the average response intensities to the two peptides in the complete response patients were obiviously higher than the patients (who had not a good curative effect) after antiviral therapy (P<0.05). But there was no statistically significance between the two groups before the treatment (P> 0.05). Intra-group comparison: the rates of response and the average response intensities to the two peptides in the complete response patients were obiviously higher than before the treatment (P<0.05). But there was no statistically significance in the patients (who had not a good curative) before and after treatment (P> 0.05).Conclusion1. There was no statistically significance the distribution of HLA-A2 gene frequency among the four groups.2. The secretion level of IFN-γis different, when PBMC with the different behavior in HBV infection under the stimulation of (HBcAgl8-27V/I) .This study showed that the cellular immune response to HBcAg was much higher in covert infection group, even the virus has been clearanced for long time. These demonstrated HBV specific CTL might play might play an important role in clearance the virus.3. The patients who received antiviral therapy could recovered the immune response,and also enhanced the ability of peripheral blood lymphocytes produces IFN-γ.4. The method to detect ability of T cell produce IFN-γby ELISPOT could be a useful cellular immune marker for evaluating the ability of anti-virus in HBV infection.