Objective: To screen and identify the hepatitis B virus (HBV) antigenic T cell epitopes restricted by human leukocyte antigen (HLA)-A0201, A1101 and A2402 which have a total gene frequency of >50% in Chinese population. Methods: Six online data banks for T cell epitopes prediction were used to virtually screen the T cell epitopes derived from HBsAg, HBcAg, HBpol and HBx proteins and restricted by the three HLA-A molecules,respectively. Peripheral anticoagulation samples from HBV-infected in-patients were collected from the Department of Clinical Laboratory of Nanjing the Second Hospital, and peripheral blood mononuclear cells (PBMCs) were processed.Then the interferon-γ enzyme linked immunospot (IFN-γ-ELISPOT) assay was performed to screen out the patients who had the specific T cell responses to the cocktail of peptides. The fresh peripheral blood samples were collected again from these patients and the immunogenicity of each peptide was confirmed by the IFN-γ-ELISPOT assay. Meanwhile, HLA-A genotyping was carried out in these HBV-infected patients through polymerase chain reaction-sequencing-based typing (PCR-SBT) method. Results: The immunogenicity of eight epitopes restricted by HLA-A0201, six epitopes restricted by A1101, and seven epitopes restricted by A2402 were confirmed by functional experiment. Conclusion: Twenty-one T cell epitopes have been identified, which derived from four kinds of HBV proteins and restricted by three HLA-A molecules, respectively. Of them, eleven epitopes have not been reported before. Moreover, the immunogenicity of the epitopes restricted by HLA-A1101 is significantly weaker than that restricted by HLA-A0201 and A2402.
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