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Journal of Virology, May 1999, p. 4508-4512, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Detection of Simian Immunodeficiency Virus Gag-Specific CD8+ T Lymphocytes in Semen of Chronically Infected Rhesus Monkeys by Cell Staining with a Tetrameric Major Histocompatibility Complex Class I-Peptide Complex

Holly L. Jordan,* Marcelo J. Kuroda, Jörn E. Schmitz, Tavis Steenbeke, Meryl A. Forman,dagger and Norman L. Letvin

Division of Viral Pathogenesis, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts 02215

Received 9 November 1998/Accepted 14 February 1999

Evaluation of human immunodeficiency virus type 1-specific mucosal cytotoxic T lymphocytes can be hampered by limited cell yields from mucosal sites. We sought to characterize virus-specific CD8+ T lymphocytes with cytotoxic activity in the male genital tracts of SIVmac-infected rhesus monkeys by using a peptide epitope-specific functional T-cell assay and a tetrameric major histocompatibility complex class I-peptide complex. This tetrameric complex was constructed with the rhesus monkey HLA-A homolog molecule Mamu-A*01 and a dominant-epitope 9-amino-acid fragment of SIVmac Gag (p11C, C-M). The proportion of tetramer-positive CD8+ T cells in semen of SIVmac-infected monkeys ranged from 5.9 to 22.0%. By the use of a standard 51Cr release assay, these cells were found to have peptide epitope-specific cytolytic activity after in vitro expansion. Four-color flow-cytometric analysis of these seminal tetramer-positive CD8+ T cells demonstrated that they express memory-associated (CD62L- CD45RA-) and activation-associated (CD11a+ Fas+ HLA-DR+) molecules. The present experiments illustrate the power of tetramer technology for evaluating antigen-specific CD8+ T lymphocytes in a mucosal tissue compartment.

* Corresponding author. Mailing address: Division of Infectious Diseases, Department of Medicine, 547 Burnett-Womack Building, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599-7030. Phone: (919) 513-2819. Fax: (919) 966-6714. E-mail: holly_jordan{at}med.unc.edu.

dagger Present address: Beckman Coulter, Inc., Miami, FL 33196.

Journal of Virology, May 1999, p. 4508-4512, Vol. 73, No. 5
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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