Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Oct;83(19):9803-12.
doi: 10.1128/JVI.00776-09. Epub 2009 Jul 29.

Diverse cross-reactive potential and Vbeta gene usage of an epitope-specific cytotoxic T-lymphocyte population in monkeys immunized with diverse human immunodeficiency virus type 1 Env immunogens

Sandrine L Hulot  1 , Michael S Seaman, Pritha Sen, Patrick A Autissier, Edwin R Manuel, Norman L Letvin
Affiliations

Diverse cross-reactive potential and Vbeta gene usage of an epitope-specific cytotoxic T-lymphocyte population in monkeys immunized with diverse human immunodeficiency virus type 1 Env immunogens

Sandrine L Hulot et al. J Virol. 2009 Oct.

Abstract

An ideal human immunodeficiency virus type 1 (HIV-1) vaccine would elicit potent cellular and humoral immune responses that recognize diverse strains of the virus. In the present study, combined methodologies (flow cytometry, Vbeta repertoire analysis, and complementarity-determining region 3 sequencing) were used to determine the clonality of CD8(+) T lymphocytes taking part in the recognition of variant epitope peptides elicited in Mamu-A*01-positive rhesus monkeys immunized with vaccines encoding diverse HIV-1 envelopes (Envs). Monkeys immunized with clade B Envs generated CD8(+) T lymphocytes that cross-recognized both clade B- and clade C-p41A epitope peptides using a large degree of diversity in Vbeta gene usage. However, with two monkeys immunized with clade C Env, one monkey exhibited p41A-specific cytotoxic T-lymphocytes (CTL) with the capacity for cross-recognition of variant epitopes, while the other monkey did not. These studies demonstrate that the cross-reactive potential of variant p41A epitope peptide-specific CTL populations can differ between monkeys that share the same restricting major histocompatibility complex class I molecule and receive the same vaccine immunogens.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
Cross-reactivity of p41A epitope-specific CD8+ T-lymphocyte populations in 89.6P Env-immunized Mamu-A*01+ rhesus monkeys. PBMC were isolated from monkeys AW13, AW28, and AW2P following vaccination. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 with HxB2-p41A peptide (A), 89.6P-p41A peptide (B), or clade C-p41A peptide (C) and then double stained with HxB2-p41A and 89.6P- p41A tetramers, clade C-p41A and 89.6P-p41A tetramers, or HxB2-p41A and clade C-p41A tetramers. Data presented are dot plots gated on CD3+ CD8+ lymphocytes, with the percentages of gated cells in each quadrant indicated. PE, phycoerythrin; APC, allophycocyanin.
FIG. 2.
FIG. 2.
Cross-reactivity of p41A epitope-specific CD8+ T-lymphocyte populations in 89.6P Env-immunized Mamu-A*01+ rhesus monkeys following SHIV-89.6P challenge. PBMC were isolated from monkeys AW13, AW28, and AW2P on day 142 following SHIV-89.6P challenge. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 with HxB2-p41A peptide (A), 89.6P-p41A peptide (B), or clade C-p41A peptide (C) and then double stained with HxB2-p41A and 89.6P-p41A tetramers, clade C-p41A and 89.6P-p41A tetramers, or HxB2-p41A and clade C-p41A tetramers. Data presented are dot plots gated on CD3+ CD8+ lymphocytes, with the percentages of gated cells in each quadrant indicated. PE, phycoerythrin; APC, allophycocyanin.
FIG. 3.
FIG. 3.
Vβ repertoire analysis of p41A epitope-specific CD8+ T-lymphocyte populations in 89.6P Env-immunized Mamu-A*01+ rhesus monkeys following vaccination and challenge. PBMC were isolated from monkeys AW13, AW28, and AW2P following vaccination and on day 142 following SHIV-89.6P challenge. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 with HxB2-p41A peptide, clade C-p41A peptide, or 89.6P-p41A peptide and then double stained with HxB2-p41A and 89.6P-p41A tetramers, clade C-p41A and 89.6P-p41A tetramers, or HxB2-p41A and clade C-p41A tetramers. HxB2-p41A-, clade C-p41A-, and 89.6P-p41A-specific CD8+ T-lymphocyte populations were sorted. The RNA isolated from sorted p41A-specific T cells (CD3+, CD8+, and tetramer positive) was used to generate cDNA for Vβ repertoire analysis performed by real-time PCR using a Cβ primer and 46 primers specific for Vβ gene families. Data are presented as the percentages of total Vβ ([{number of copies of a Vβ gene family − background}/{number of copies of all Vβ gene families − background}] × 100) for each Vβ gene family.
FIG. 4.
FIG. 4.
Cross-reactivity of p41A epitope-specific CD8+ T-lymphocyte populations in HxB2 Env-immunized Mamu-A*01+ rhesus monkeys following vaccination and challenge. PBMC were isolated from monkeys 419 and VFA at week 27 following vaccination (1-week rAd boost) or week 28 following SHIV-89.6P challenge. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 with HxB2-p41A peptide or 89.6P-p41A peptide and then double stained with HxB2-p41A and 89.6P-p41A tetramers. Data presented are dot plots gated on CD3+ CD8+ lymphocytes, with the percentages of gated cells in each quadrant indicated. PE, phycoerythrin; APC, allophycocyanin.
FIG. 5.
FIG. 5.
p41A-specific IFN-γ ELISPOT responses of PBMC from Env-immunized Mamu-A*01+ rhesus monkeys. (A) Responses observed for HxB2 Env-immunized monkeys (VFA and 419). (B) Responses observed for clade C Env-immunized monkeys (414 and KPA). PBMC were isolated from monkeys VFA, 419, KPA, and 414 at week 27 following vaccination (1-week post-rAd boost). Cells were stimulated in vitro in the presence of media alone or the noted p41A variant peptides, and IFN-γ SFC responses were measured. The mean numbers of spots per 106 PBMC are shown.
FIG. 6.
FIG. 6.
Vβ repertoire analysis of p41A epitope-specific CD8+ T-lymphocyte populations in HxB2 Env-immunized Mamu-A*01+ rhesus monkeys following vaccination and challenge. PBMC were isolated from monkeys 419 and VFA at week 27 following vaccination or week 28 following SHIV-89.6P challenge. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 with HxB2-p41A peptide or 89.6P-p41A peptide. RNA isolated from sorted p41A-specific T cells (CD3+, CD8+, and tetramer positive) was used to generate cDNA for Vβ repertoire analysis by real-time PCR using a Cβ primer and 46 primers specific for Vβ gene families. Data are presented as the percentages of total Vβ gene families for each Vβ family.
FIG. 7.
FIG. 7.
Cross-reactivity of p41A epitope-specific CD8+ T-lymphocyte populations in clade C Env-immunized Mamu-A*01+ rhesus monkeys following vaccination and challenge. PBMC were isolated from monkeys KPA and 414 at week 27 following vaccination (1-week post-rAd boost) or week 20 following SHIV-89.6P challenge. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 with clade C-p41A peptide or 89.6P-p41A peptide and then double stained with clade C-p41A and 89.6P-p41A tetramers. Data presented are dot plots gated on CD3+ CD8+ lymphocytes, with the percentages of gated cells in each quadrant indicated. PE, phycoerythrin; APC, allophycocyanin.
FIG. 8.
FIG. 8.
Vβ repertoire analysis of p41A epitope-specific CD8+ T-lymphocyte populations in clade C Env-immunized Mamu-A*01+ rhesus monkeys following vaccination and challenge. PBMC were isolated from monkeys KPA and 414 at week 27 following vaccination or week 20 following SHIV-89.6P challenge. Cells were stimulated in vitro for 12 to 14 days in the presence of IL-2 and clade C-p41A peptide or 89.6P-p41A peptide. RNA isolated from sorted p41A-specific T cells (CD3+, CD8+, and tetramer positive) was used to generate cDNA for Vβ repertoire analysis by real-time PCR using a Cβ primer and 46 primers specific for Vβ gene families. Data are presented as the percentages of total Vβ gene families for each Vβ gene family.
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Allen, T. M., D. H. O'Connor, P. Jing, J. L. Dzuris, B. R. Mothe, T. U. Vogel, E. Dunphy, M. E. Liebl, C. Emerson, N. Wilson, K. J. Kunstman, X. Wang, D. B. Allison, A. L. Hughes, R. C. Desrosiers, J. D. Altman, S. M. Wolinsky, A. Sette, and D. I. Watkins. 2000. Tat-specific cytotoxic T lymphocytes select for SIV escape variants during resolution of primary viraemia. Nature 407:386-390. - PubMed
    1. Appay, V., D. C. Douek, and D. A. Price. 2008. CD8+ T cell efficacy in vaccination and disease. Nat. Med. 14:623-628. - PubMed
    1. Barouch, D. H., A. Craiu, S. Santra, M. A. Egan, J. E. Schmitz, M. J. Kuroda, T. M. Fu, J. H. Nam, L. S. Wyatt, M. A. Lifton, G. R. Krivulka, C. E. Nickerson, C. I. Lord, B. Moss, M. G. Lewis, V. M. Hirsch, J. W. Shiver, and N. L. Letvin. 2001. Elicitation of high-frequency cytotoxic T-lymphocyte responses against both dominant and subdominant simian-human immunodeficiency virus epitopes by DNA vaccination of rhesus monkeys. J. Virol. 75:2462-2467. - PMC - PubMed
    1. Barouch, D. H., J. Powers, D. M. Truitt, M. G. Kishko, J. C. Arthur, F. W. Peyerl, M. J. Kuroda, D. A. Gorgone, M. A. Lifton, C. I. Lord, V. M. Hirsch, D. C. Montefiori, A. Carville, K. G. Mansfield, K. J. Kunstman, S. M. Wolinsky, and N. L. Letvin. 2005. Dynamic immune responses maintain cytotoxic T lymphocyte epitope mutations in transmitted simian immunodeficiency virus variants. Nat. Immunol. 6:247-252. - PubMed
    1. Borrow, P., H. Lewicki, B. H. Hahn, G. M. Shaw, and M. B. Oldstone. 1994. Virus-specific CD8+ cytotoxic T-lymphocyte activity associated with control of viremia in primary human immunodeficiency virus type 1 infection. J. Virol. 68:6103-6110. - PMC - PubMed

Publication types