Identification of human CD4+ T-cell epitopes on the VP1 capsid protein of enterovirus 71

Abstract

The identification of human CD4+ T-cell epitopes within a protein vaccine candidate is of great interest, as it provides a better understanding of the mechanisms involved in protective immunity and may therefore help in the design of effective vaccines and diagnostic tools. The entire amino acid sequence of the VP1 capsid protein from enterovirus 71 (EV71) strain 41 was submitted to analysis by the ProPred algorithm for the identification of potential promiscuous human CD4+ T-cell epitopes. Three regions spanning amino acids 66-77, 145-159, and 247-261 of VP1 were predicted to bind more than 25 HLA-DR alleles. The corresponding synthetic peptides (SP1 to SP3) were then tested for their abilities to induce proliferation of CD4+ T cells isolated from five human volunteers screened positive for previous EV71 exposure and one EV71-negative volunteer. Upon stimulation with either peptide, CD4+ T-cell proliferative responses were observed for all EV71-positive volunteers, indicating the presence of EV71-specific memory CD4+ T cells. The amplitude of the proliferative responses was peptide- and HLA-DR-dependent, and correlated well with the ProPred predicted binding efficiencies. Moreover, CD4+T cells from EV71-positive volunteers produced significant levels of IL-2 and IFN-γ upon stimulation, indicative of a T-cell differentiation into Th-1-type subset. Among the three peptides, SP2 induced the highest proliferative response and cytokine production. Moreover, SP2-induced proliferative response could be inhibited with anti-major his-tocompatibility complex (MHC) class II antibody, indicating that SP2 represents a MHC class II-restricted CD4+ T-cell epitope. This study demonstrates that the ProPred algorithm can accurately predict the presence of human CD4+ T-cell epitopes within the VP1 capsid protein of EV71, and therefore represents a useful tool for the design of subunit vaccines against EV71. © Mary Ann Liebert, Inc. 2008.