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Journal of Virology, February 2003, p. 2081-2092, Vol. 77, No. 3
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.3.2081-2092.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Comprehensive Epitope Analysis of Human Immunodeficiency Virus Type 1 (HIV-1)-Specific T-Cell Responses Directed against the Entire Expressed HIV-1 Genome Demonstrate Broadly Directed Responses, but No Correlation to Viral Load

M. M. Addo,1 X. G. Yu,1 A. Rathod,1 D. Cohen,2 R. L. Eldridge,1 D. Strick,1 M. N. Johnston,1 C. Corcoran,1 A. G. Wurcel,1 C. A. Fitzpatrick,1 M. E. Feeney,1 W. R. Rodriguez,1 N. Basgoz,1 R. Draenert,1 David R. Stone,3 C. Brander,1 P. J. R. Goulder,1,4 E. S. Rosenberg,1 M. Altfeld,1 and B. D. Walker1*

Partners AIDS Research Center, Massachusetts General Hospital and Harvard Medical School,1 Fenway Community Health Center,2 Lemuel Shattuck Hospital, Boston, Massachusetts,3 Nuffield Department of Medicine, John Radcliffe Hospital, Oxford, United Kingdom4

Received 19 August 2002/ Accepted 5 November 2002

Cellular immune responses play a critical role in the control of human immunodeficiency virus type 1 (HIV-1); however, the breadth of these responses at the single-epitope level has not been comprehensively assessed. We therefore screened peripheral blood mononuclear cells (PBMC) from 57 individuals at different stages of HIV-1 infection for virus-specific T-cell responses using a matrix of 504 overlapping peptides spanning all expressed HIV-1 proteins in a gamma interferon-enzyme-linked immunospot (Elispot) assay. HIV-1-specific T-cell responses were detectable in all study subjects, with a median of 14 individual epitopic regions targeted per person (range, 2 to 42), and all 14 HIV-1 protein subunits were recognized. HIV-1 p24-Gag and Nef contained the highest epitope density and were also the most frequently recognized HIV-1 proteins. The total magnitude of the HIV-1-specific response ranged from 280 to 25,860 spot-forming cells (SFC)/106 PBMC (median, 4,245) among all study participants. However, the number of epitopic regions targeted, the protein subunits recognized, and the total magnitude of HIV-1-specific responses varied significantly among the tested individuals, with the strongest and broadest responses detectable in individuals with untreated chronic HIV-1 infection. Neither the breadth nor the magnitude of the total HIV-1-specific CD8+-T-cell responses correlated with plasma viral load. We conclude that a peptide matrix-based Elispot assay allows for rapid, sensitive, specific, and efficient assessment of cellular immune responses directed against the entire expressed HIV-1 genome. These data also suggest that the impact of T-cell responses on control of viral replication cannot be explained by the mere quantification of the magnitude and breadth of the CD8+-T-cell response, even if a comprehensive pan-genome screening approach is applied.

* Corresponding author. Mailing address: MGH-East, 149 13th St., Charlestown, MA 02129. Phone: (617) 724-8332. Fax: (617) 726-4691. E-mail: bwalker{at}partners.org.

Journal of Virology, February 2003, p. 2081-2092, Vol. 77, No. 3
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.3.2081-2092.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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