Screening and identification of HLA-A0201 restricted CTL epitope from hepatitis B virus X protein in vitro / 第二军医大学学报

ABSTRACT

Objective: To identify the HLA-A0201 restricted CTL epitopes derived from hepatitis B virus X protein predicted by computer program and general principles in vitro. Methods: HBx gene sequences of Hepatitis B virus genotypes B/C and serotypes adw/adr, with the highest frequencies in Chinese, were computed and analyzed by screening service offered by Internet combined with peptide supermotif, extended motif and quantitative motif prediction. Four most ideal nine-peptides (HBx1, HBx2, HB3x3, and HBx4) were selected as candidate peptides. Using flow cytometry, the fluorescence index of both control and experimental groups were detected and the 4 nine-peptides were evaluated with T2 binding assay and DC50 assay. Results: The nine-peptides VLCLRPVGA (HBx1), CLFKDWEEL (HBx2), VLHKRTLGL (HBx3) and HLSLRGLPV (HBx4) were selected as candidate targets. Among the 4 candidate peptides, HBx2 showed higher HLA-A0201 affinity and HBx2, HBx4 showed better stability. Conclusion: Our study indicates that CLFKDWEEL might be a potential HLA-A0201 restricted CTL epitope from hepatitis B virus X protein; further study is needed for verification of its immunity in vivo.