Identification of an HLA-A*0201-restricted CD8+ T-cell epitope SSp-1 of SARS-CoV spike protein - PubMed (original) (raw)
. 2004 Jul 1;104(1):200-6.
doi: 10.1182/blood-2003-11-4072. Epub 2004 Mar 11.
Huabiao Chen, Xiaodong Jiang, Minghui Zhang, Tao Wan, Nan Li, Xiangyang Zhou, Yanfeng Wu, Feng Yang, Yizhi Yu, Xiaoning Wang, Ruifu Yang, Xuetao Cao
Affiliations
- PMID: 15016646
- PMCID: PMC8254376
- DOI: 10.1182/blood-2003-11-4072
Identification of an HLA-A*0201-restricted CD8+ T-cell epitope SSp-1 of SARS-CoV spike protein
Baomei Wang et al. Blood. 2004.
Abstract
A novel coronavirus, severe acute respiratory syndrome (SARS)-associated coronavirus (SARS-CoV), has been identified as the causal agent of SARS. Spike (S) protein is a major structural glycoprotein of the SARS virus and a potential target for SARS-specific cell-mediated immune responses. A panel of S protein-derived peptides was tested for their binding affinity to HLA-A*0201 molecules. Peptides with high affinity for HLA-A*0201 were then assessed for their capacity to elicit specific immune responses mediated by cytotoxic T lymphocytes (CTLs) both in vivo, in HLA-A2.1/K(b) transgenic mice, and in vitro, from peripheral blood lymphocytes (PBLs) harvested from healthy HLA-A2.1(+) donors. SARS-CoV protein-derived peptide-1 (SSp-1 RLNEVAKNL), induced peptide-specific CTLs both in vivo (transgenic mice) and in vitro (human PBLs), which specifically released interferon-gamma (IFN-gamma) upon stimulation with SSp-1-pulsed autologous dendritic cells (DCs) or T2 cells. SSp-1-specific CTLs also lysed major histocompatibility complex (MHC)-matched tumor cell lines engineered to express S proteins. HLA-A*0201-SSp-1 tetramer staining revealed the presence of significant populations of SSp-1-specific CTLs in SSp-1-induced CD8(+) T cells. We propose that the newly identified epitope SSp-1 will help in the characterization of virus control mechanisms and immunopathology in SARS-CoV infection, and may be relevant to the development of immunotherapeutic approaches for SARS.
Figures
Figure 1
Immunogenicity of SARS-S–derived peptide SSp-1 in HLA-A2.1/Kb transgenic mice. IFN-γ release by bulk CTLs from immunized mice was detected by ELISPOT assay (A) and lytic activity was tested using a standard 4-hour chromium release assay (B). (A) Bulk CTLs from SSp-1–immunized mice released IFN-γ in response to T2 cells pulsed with SSp-1 (T2/SSp-1) in a dose-dependent manner, but not those with irrelevant peptide CAP-1 (T2/CAP-1) or T2 cells alone. (B) Bulk CTLs from SSp-1–immunized mice lysed T2 cells loaded with SSp-1, but not T2 cells pulsed with the irrelevant peptide CAP-1 or T2 cells alone. Data represent means ± SD.
Figure 2
Specific reactivity of human CTLs induced by SSP-1–loaded DCs in vitro. SSp-1–specific CTLs were generated from the PBMCs of 7 of 11 HLA-A2.1+ healthy donors through 4 sequential rounds of stimulation with SSp-1–pulsed DCs. Resulting CTLs were tested for IFN-γ release (A-B) and SSp-1–specific lysis (C) using an ELISPOT assay and a standard 4-hour chromium release assay. Autologous DCs (A) and T2 cells (B) were pulsed with indicated concentrations of SSp-1 (DC/SSp-1, T2/SSp-1) or irrelevant peptide CAP-1 (DC/CAP-1, T2/CAP-1), and then used as stimulators in an IFN-γ release assay. (C) CTLs lysed T2 cells loaded with SSp-1, but not T2 cells loaded with the irrelevant peptide CAP-1 or T2 cells alone. Results are representative of 3 independent experiments. Data represent means ± SD.
Figure 3
Identification of SSp-1 as a naturally processed and presented HLA-A2.1–restricted epitope. DC-induced human CTLs against SSp-1 were tested for specific lytic activity (A) and IFN-γ release (B) in response to genetically modified cell lines. (A) CTLs lysed pAd-SARS/S–transduced HLA-A2.1+ SW480 cells (SW480/SARS/S) but did not lyse pAd-SARS/S–transduced HLA-A2.1– HT29 cells (HT29/SARS/S) or pAd-LacZ–transduced SW480 (SW480/LacZ) and HT29 cells (HT29/LacZ). (B) CTLs elicited strong IFN-γ production in response to pAd-SARS/S–transduced SW480 cells, but only very low levels were elicited by pAd-SARS/S–transduced HT29 cells or pAd-LacZ–transduced cell lines. Results are representative of 3 independent experiments. Data represent means ± SD.
Figure 4
Frequency of SSp-1–specific CD8+ T cells. SSp-1–specific CD8+ T-cell frequency among DC-induced human CTLs against SSp-1 (A-B) and splenocytes derived from immunized mice and restimulated for 6 days in vitro (C-D). Tetramer-binding CD8+ T cells are shown in the upper-right quadrant and are labeled with the percentage of total CD8+ T cells. SSp-1–specific CTLs generated from human PBLs were stained with control HLA-A*0201/CAP-1 tetramers (A) and HLA-A*0201/SSp-1 tetramers (B). Results are representative of 3 independent experiments. Splenocytes from Tg mice immunized with SSp-1–pulsed DCs were stained with control HLA-A*0201/CAP-1 tetramers (C) and HLA-A*0201/SSp-1 tetramers (D).
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