Identification of HLA-A*2402-restricted HCMV immediate early-1 (IE-1) epitopes as targets for CD8+ HCMV-specific cytotox
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Identification of HLA-A*2402-restricted HCMV immediate early-1 (IE-1) epitopes as targets for CD8+ HCMV-specific cytotoxic T lymphocytes Jong-Baeck Lim1, Hyun Ok Kim1, Seok Hoon Jeong1, Joo Eun Ha1, Sunphil Jang1, Sang-Guk Lee1, Kyungwon Lee1 and David Stroncek*2 Address: 1Department of Laboratory Medicine, Yonsei University College of Medicine, Seoul, South Korea and 2Department of Transfusion Medicine, Warren G. Magnuson Clinical Center, National Institutes of Health, Bethesda, MD, USA Email: Jong-Baeck Lim - [email protected]; Hyun Ok Kim - [email protected]; Seok Hoon Jeong - [email protected]; Joo Eun Ha - [email protected]; Sunphil Jang - [email protected]; SangGuk Lee - [email protected]; Kyungwon Lee - [email protected]; David Stroncek* - [email protected] * Corresponding author
Published: 23 August 2009 Journal of Translational Medicine 2009, 7:72
doi:10.1186/1479-5876-7-72
Received: 1 June 2009 Accepted: 23 August 2009
This article is available from: http://www.translational-medicine.com/content/7/1/72 © 2009 Lim et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: To identify novel HLA-A*2402-restricted human cytomegalovirus (HCMV) immediate early-1 (IE-1) epitopes for adoptive immunotherapy, we explored 120 overlapping 15amino acid spanning IE-1. Methods: These peptides were screened by measuring the frequency of polyclonal CD8+ T cells producing intracellular interferon-γ (IFN-γ) using flow cytometry and the epitopes were validated with a HCMV-infected target Cr release cytotoxicity assay. Results: Initial screening was performed with 12 mini-pools of 10 consecutive peptides made from 120 overlapping peptides15-amino acids in length that spanned IE-1. When peripheral blood mononuclear cells (PBMCs) from HLA-A*2402 HCMV-seropositive donors were sensitized with each of the 12 mini-pools, mini-pools 1 and 2 induced the highest frequency of CD8+ cytotoxic T lymphocytes (CTLs) producing IFN-γ. When PBMCs were stimulated with each of the twenty peptides belonging to mini-pools 1 and 2, peptides IE-11–15MESSAKRKMDPDNPD and IE-15– 19AKRKMDPDNPDEGPS induced the greatest quantities of IFN-γ production and cytotoxicity of HLA-matched HCMV-infected fibroblasts. To determine the exact HLA-A*2402-restricted epitopes within the two IE-1 proteins, we synthesized a total of twenty-one overlapping 9- or 10 amino acid peptides spanning IE-11–15 and IE-15–19. Peptide IE-13–12SSAKRKMDPD induced the greatest quantities of IFN-γ production and target cell killing by CD8+ CTLs. Conclusion: HCMV IE-13–12SSAKRKMDPD is a HLA-A*2402-restricted HCMV IE-1 epitope that can serve as a common target for CD8+ HCMV-specific CTLs.
Background Human cytomegalovirus (HCMV) infe
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