| PART 1.HEPATITIS B VIRUS MUTATION AND T-CELL IMMUNITY IN HOST AND SERUM HBEAG STATUS,ALT AND HBV DNA LEVELSObjective: To investigate the relationship between HBV mutation and host T-cell immunity and serum HBe Ag status,serum ALT and HBV DNA levels.Methods: In the cross-sectional study,35 untrated patients with chronic hepatitis B(CHB)were included.HBV ORF-Pre C/C and ORF-S were analysed to screen for variants with effects on HBe Ag status,serum ALT,and HBV DNA levels by the PCR-TA cloning-sequencing method.These effects were further verified,after adjustment for potential confounders in the longitudinal study(n=60)by multiple regression analysis.Quantitative microarray was used to detect the expression levels of T cell related cytokines(IFN-?,TNF-?,IL-2,IL-17,IL-17 F,IL-22,IL-23,and IL-21)in the host.Results: In a cross-sectional study,64.4% mutation sites and 68.2% mutation sites were found in the epitope region of ORF-Pre C/C and ORF-S respectively.There are 10(Pre C/C50,55,79,84,103,126,145,184,and s110,s213),8(Pre C/C28,64,116),145,182 and s14,s21,s126),and 5 mutation sites(Pre C/C55,79,84,126,and 184)were associated with HBe Ag(-)status,abnormal ALT levels and lower HBV DNA levels,respectively.After adjustment for confounding factors such as age,gender,genotype,serum ALT levels,and Pre C W28*,the mutation of Tc cell epitopes(Pre C47-56,Pre C 117-125,s14-22,s208-216)and Th(Pre C176-185)and Pre C W28* were the independent factors for host HBe Ag status,ALT levels,and HBV DNA levels.in the longitudinal study.In addition,amino acid substitutions in the epitope region may affect the hydrophobicity and the expression of Th cell-related cytokines.Conclusion: The mutations in the HBV ORF-Pre C/C(Pre C47-56,Pre C 117-125 and Pre C176-185)and ORF-S region(s14-22,s208-216)epitopes were the main independent factor for the HBe Ag status,ALT,and HBV DNA levels in the untrated CHB patients.PART 2 HIGHER BASELINE VIRAL DIVERSITY CORRELATES WITH LOWER HBSAG DECLINE FOLLOWING PEGYLATED INTERFERON-ALPHA THERAPY IN PATIENTS WITH HBEAG-POSITIVE CHRONIC HEPATITIS BObjective: Viral diversity seems to predict treatment outcomes in certain viral infections.The aim of this study was to evaluate the association between baseline intra-patient viral diversity and HBs Ag decline following PEGylated interferon-alpha(Peg-IFN-?)therapy.Methods: Twenty-six HBe Ag positive patients who were treated with Peg-IFN-? were enrolled.Nested PCR,cloning and sequencing of the HBV S gene were performed on baseline samples,and normalized Shannon entropy(Sn)was calculated as a measure of small hepatitis B surface protein(SHBs)diversity.Multiple regression analysis was used to estimate the association between baseline Sn and HBs Ag decline.Results: Of the 26 patients enrolled in the study,65.4% were male and 61.5% were infected with HBV genotype B.The median HBs Ag level at baseline was 4.5 log10 IU/ml(interquartile range(IQR): 4.1-4.9)and declined to 3.0 log10 IU/ml(IQR: 1.7-3.9)after 48 weeks of Peg-IFN-? treatment.In models adjusted for baseline ALT and HBs Ag,the adjusted coefficients(95% confidence interval)for ?HBs Ag and relative percentage HBs Ag decrease were-1.3(-2.5,-0.2)log10 IU/ml for higher SHBs diversity(Sn ? 0.58)patients and-26.4%(-50.2%,-2.5%)for lower diversity(Sn < 0.58)patients.Further analysis showed that the “a” determinant upstream flanking region and the first loop of the “a” determinant(nucleotides 341-359,371-389 and 381-399)were the main sources of higher SHBs diversity.Conclusions: Baseline intra-patient SHBs diversity was inverse to HBs Ag decline in HBe Ag-positive CHB patients receiving Peg-IFN-? monotherapy.Moreover,more sequence variations within the “a” determinant upstream flanking region and the first loop of the “a” determinant were the main sources of the higher SHBs diversity. |
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