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. 1999 May;73(5):4508-12.
doi: 10.1128/JVI.73.5.4508-4512.1999.

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

H L Jordan  1 , M J Kuroda, J E Schmitz, T Steenbeke, M A Forman, N L Letvin
Affiliations

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

H L Jordan et al. J Virol. 1999 May.

Abstract

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.

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Figures

FIG. 1
FIG. 1
Percentages of CD3+ CD8α/β+ cells binding tetrameric Mamu-A*01–p11C, C-M complex in peripheral blood leukocytes (PBL) and freshly collected semen of an uninfected Mamu-A*01+ control rhesus monkey with leukocytic semen (SIV−) and SIVmac-infected, Mamu-A*01+ monkeys (no. 579, 138, 403, and 575) as determined by tricolor flow cytometry.
FIG. 2
FIG. 2
(Top) The percentages of tetramer-binding CD3+ CD8α/β+ cells in aliquots of cultured seminal lymphocytes from SIVmac-infected Mamu-A*01+ monkeys, assessed by flow cytometry as described in the text. (Bottom) SIVmac Gag epitope p11C, C-M-specific lytic activity in in vitro-expanded seminal lymphocytes from these monkeys. E/T, effector/target.
FIG. 3
FIG. 3
Phenotypic characterization of tetrameric Mamu-A*01–p11C, C-M complex-negative (Tetramer−) and complex-positive (Tetramer+) CD8α/β+ T cells in freshly collected semen from an SIVmac-infected Mamu-A*01+ monkey (no. 579). Flow cytometric analysis was performed on gated CD3+ CD8α/β+ T cells.
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