Persistent memory CD4+ and CD8+ T-cell responses in recovered severe acute respiratory syndrome (SARS) patients to SARS coronavirus M antigen

Abstract
The membrane (M) protein of severe acute respiratory syndrome coronavirus (SARS-CoV) is a major glycoprotein with multiple biological functions. In this study, we found that memory T cells against M protein were persistent in recovered SARS patients by detecting gamma interferon (IFN-γ) production using ELISA and ELISpot assays. Flow cytometric analysis showed that both CD4+and CD8+T cells were involved in cellular responses to SARS-CoV M antigen. Furthermore, memory CD8+T cells displayed an effector memory cell phenotype expressing CD45ROCCR7CD62L. In contrast, the majority of IFN-γ+CD4+T cells were central memory cells with the expression of CD45RO+CCR7+CD62L. The epitope screening from 30 synthetic overlapping peptides that cover the entire SARS-CoV M protein identified four human T-cell immunodominant peptides, p21-44, p65-91, p117-140 and p200-220. All four immunodominant peptides could elicit cellular immunity with a predominance of CD8+T-cell response. This data may have important implication for developing SARS vaccines.