Rapid definition of five novel HLA-A*3002-Restricted human immunodeficiency virus-specific cytotoxic T-lymphocyte epitopes by elispot and intracellular cytokine staining assays

P.J.R. Goulder; M.M. Addo; M.A. Altfeld; E.S. Rosenberg; Y. Tang; U. Govender; N. Mngqundaniso; K. Annamalai; T.U. Vogel; M. Hammond; M. Bunce; H.M. Coovadia; B.D. Walker

doi:10.1128/JVI.75.3.1339-1347.2001

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Rapid definition of five novel HLA-A*3002-Restricted human immunodeficiency virus-specific cytotoxic T-lymphocyte epitopes by elispot and intracellular cytokine staining assays

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Rapid definition of five novel HLA-A*3002-Restricted human immunodeficiency virus-specific cytotoxic T-lymphocyte epitopes by elispot and intracellular cytokine staining assays

P.J.R. Goulder, M.M. Addo, M.A. Altfeld, E.S. Rosenberg, Y. Tang, U. Govender, N. Mngqundaniso, K. Annamalai, T.U. Vogel, M. Hammond, …

Journal of Virology, Vol.75(3), pp.1339-1347

2001

DOI: https://doi.org/10.1128/JVI.75.3.1339-1347.2001

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Abstract

Human immunodeficiency virus (HIV)-specific cytotoxic T lymphocytes (CTL) play a major role in control of viral replication. To understand the contribution of this antiviral response, an initial step is to fully define the specific epitopes targeted by CTL. These studies focused on CTL responses restricted by HLA-A*3002, one of the HLA-A molecules most prominent in African populations. To avoid the time-consuming effort and expense involved in culturing CTL prior to defining epitopes and restricting alleles, we developed a method combining Elispot assays with intracellular gamma interferon staining of peripheral blood mononuclear cells to first map the optimal epitopes targeted and then define the HLA restriction of novel epitopes. In two A*3002-positive subjects whose CTL responses were characterized in detail, the strongest response in both cases was to an epitope in p17 Gag, RSLYNTVATLY (residues 76 to 86). Using this method, CTL epitopes for which there were no motif predictions were optimized and the HLA restriction was established within 48 to 72 h of receipt of blood. This simple and convenient approach should prove useful especially in the characterization of CTL responses specific to HIV and other viruses, particularly in localities where performing cytotoxicity assays would be problematic.

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Title: Rapid definition of five novel HLA-A*3002-Restricted human immunodeficiency virus-specific cytotoxic T-lymphocyte epitopes by elispot and intracellular cytokine staining assays
Authors/Creators: P.J.R. Goulder (Author/Creator) - Harvard Medical School
M.M. Addo (Author/Creator) - Harvard Medical School
M.A. Altfeld (Author/Creator) - Harvard Medical School
E.S. Rosenberg (Author/Creator) - Harvard Medical School
Y. Tang (Author/Creator) - Harvard Medical School
U. Govender (Author/Creator) - University of Natal
N. Mngqundaniso (Author/Creator) - University of Natal
K. Annamalai (Author/Creator) - University of Natal
T.U. Vogel (Author/Creator) - Wisconsin Regional Primate Center, Madison, Wisconsin 537154; and
M. Hammond (Author/Creator) - Malawi Blood Transfusion Service
M. Bunce (Author/Creator) - Churchill Hospital
H.M. Coovadia (Author/Creator) - University of Natal
B.D. Walker (Author/Creator) - Harvard Medical School
Publication Details: Journal of Virology, Vol.75(3), pp.1339-1347
Publisher: American Society for Microbiology
Identifiers: 991005544110807891
Copyright: 2001 American Society for Microbiology
Murdoch Affiliation: Murdoch University
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.66 HIV; 1.66.46 HIV Pathogenesis
Web Of Science research areas: Virology
ESI research areas: Microbiology