Abstract
Antigen-specific CD4(+) T cells play an essential role in effective immunity against Helicobacter pylori (H. pylori) infection. Lpp20, a conserved lipoprotein of H. pylori, has been investigated as one of major protective antigens for vaccination strategies. Our previous study identified two H-2(d)-restricted CD4(+) T cell epitopes within Lpp20 and an epitope vaccine based on these epitopes was constructed, which protected mice in prophylactic and therapeutic vaccination against H. pylori infection. Immunodominant CD4(+) T cell response is an important feature of antiviral, antibacterial, and antitumor cellular immunity. However, while many immunodominant HLA-restricted CD4(+) T cell epitopes of H. pylori protective antigens have been identified, immunodominant HLA-restricted Lpp20 CD4(+) T cell epitope has not been elucidated. In this study, a systematic method was used to comprehensively evaluate the immunodominant Lpp20-specific CD4(+) T cell response in H. pylori-infected patients. Using in vitro recombinant Lpp20 (rLpp20)-specific expanded T cell lines from H. pylori-infected subjects and 27 18mer overlapping synthetic peptides spanned the whole Lpp20 protein, we have shown that L(55-72) and L(79-96) harbored dominant epitopes for CD4(+) T cell responses. Then the core sequence within these two 18mer dominant epitopes was screened by various extended or truncated 13mer peptides. The immunodominant epitope was mapped to L(57-69) and L(83-95). Various Epstein-Barr virus (EBV) transformed B lymphoblastoid cell lines (B-LCLs) with different HLA alleles were used as antigen presenting cell (APC) to present peptides to CD4(+) T cells. The restriction molecules were determined by HLA class-antibody blocking. L(57-69) was restricted by DRB1-1501 and L(83-95) by DRB1-1602. The epitopes were recognized on autologous dendritic cells (DCs) loaded with rLpp20 but also those pulsed with whole cell lysates of H. pylori (HP-WCL), suggesting that these epitopes are naturally processed and presented by APC. CD4(+) T cells were isolated from H. pylori-infected patients and stimulated with L(57-69) and L(83-95). These two epitopes were able to stimulate CD4(+) T cell proliferation. This study may be of value for the future development of potential H. pylori vaccine.