Identification of Two Lpp20 CD4+ T Cell Epitopes in Helicobacter pylori-Infected Subjects

Yunshan Ning; Jianbin Ye; Junjie Wen; Danlin Wu; Zhongbiao Chen; Yanqing Lin; Bingxin Hu; Meiqun Luo; Jun Luo; Lijun Ning; Yan Li

doi:10.3389/fmicb.2018.00884

Identification of Two Lpp20 CD4⁺ T Cell Epitopes in Helicobacter pylori-Infected Subjects

Front Microbiol. 2018 May 23:9:884. doi: 10.3389/fmicb.2018.00884. eCollection 2018.

Authors

Yunshan Ning¹, Jianbin Ye¹, Junjie Wen¹, Danlin Wu¹, Zhongbiao Chen², Yanqing Lin¹, Bingxin Hu¹, Meiqun Luo¹, Jun Luo³, Lijun Ning¹, Yan Li¹

Affiliations

¹ School of Laboratory Medicine and Biotechnology, Southern Medical University, Guangzhou, China.
² Affiliated Foshan Hospital of Southern Medical University, Foshan, China.
³ School of Public Health, Southern Medical University, Guangzhou, China.

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.

Keywords: CD4+ T cell; HLA restriction; Helicobacter pylori; Lpp20; immunodominant epitope.