The Journal of Experimental Medicine
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Published online 3 July 2000.
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© The Rockefeller University Press, 0022-1007/2000/7/63/ $5.00
The Journal of Experimental Medicine, Volume 192, Number 1, July 3, 2000 63-76

Original Article

HIV-Specific Cd8+ T Cells Produce Antiviral Cytokines but Are Impaired in Cytolytic Function

Victor Appaya, Douglas F. Nixonc, Sean M. Donahoec, Geraldine M.A. Gillespiea, Tao Donga, Abigail Kinga, Graham S. Ogga, Hans M.L. Spiegelc, Christopher Conlonb, Celsa A. Spinad,e, Diane V. Havlird, Douglas D. Richmand,e, Anele Watersf, Philippa Easterbrookf, Andrew J. McMichaela, and Sarah L. Rowland-Jonesa

a Medical Research Council Human Immunology Unit, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom
b Nuffield Department of Medicine, John Radcliffe Hospital, Oxford OX3 9DU, United Kingdom
c Aaron Diamond AIDS Research Center, The Rockefeller University, New York, New York 10016
d University of California at San Diego, La Jolla, California 92093-0679
e Veterans Administration Research Center for AIDS and HIV Infection, San Diego, California 92063
f The Caldecot Centre, King's Healthcare National Health Service Trust, London SE5 9RS, United Kingdom
MRC Human Immunology Unit, Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DS, UK.44-1865-222-50244-1865-222-336

andrew.mcmichael{at}ndm.ox.ac.uk

The use of peptide–human histocompatibility leukocyte antigen (HLA) class I tetrameric complexes to identify antigen-specific CD8+ T cells has provided a major development in our understanding of their role in controlling viral infections. However, questions remain about the exact function of these cells, particularly in HIV infection. Virus-specific cytotoxic T lymphocytes exert much of their activity by secreting soluble factors such as cytokines and chemokines. We describe here a method that combines the use of tetramers and intracellular staining to examine the functional heterogeneity of antigen-specific CD8+ T cells ex vivo. After stimulation by specific peptide antigen, secretion of interferon (IFN)-{gamma}, tumor necrosis factor (TNF)-{alpha}, macrophage inflammatory protein (MIP)-1β, and perforin is analyzed by FACS® within the tetramer-positive population in peripheral blood. Using this method, we have assessed the functional phenotype of HIV-specific CD8+ T cells compared with cytomegalovirus (CMV)-specific CD8+ T cells in HIV chronic infection. We show that the majority of circulating CD8+ T cells specific for CMV and HIV antigens are functionally active with regards to the secretion of antiviral cytokines in response to antigen, although a subset of tetramer-staining cells was identified that secretes IFN-{gamma} and MIP-1β but not TNF-{alpha}. However, a striking finding is that HIV-specific CD8+ T cells express significantly lower levels of perforin than CMV-specific CD8+ T cells. This lack of perforin is linked with persistent CD27 expression on HIV-specific cells, suggesting impaired maturation, and specific lysis ex vivo is lower for HIV-specific compared with CMV-specific cells from the same donor. Thus, HIV-specific CD8+ T cells are impaired in cytolytic activity.

Key Words: cytotoxic T lymphocytes • HIV • cytokines • tetramers • perforin

Abbreviations used in this paper: APC, allophycocyanin; ELISPOT, enzyme-linked immunospot; HAART, highly active antiretroviral therapy; LCMV, lymphocytic choriomeningitis virus; LDA, limiting dilution assay; MIP, macrophage inflammatory protein; PerCP, peridinin chlorophyll protein; RANTES, regulated on activation, normal T cell expressed and secreted.

© 2000 The Rockefeller University Press


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