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      In Situ Staining and Laser Capture Microdissection of Lymph Node Residing SIV Gag-Specific CD8 + T cells—A Tool to Interrogate a Functional Immune Response Ex Vivo

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          Abstract

          While a plethora of data describes the essential role of systemic CD8 + T cells in the control of SIV replication little is known about the local in situ CD8 + T cell immune responses against SIV at the intact tissue level, due to technical limitations. In situ staining, using GagCM9 Qdot 655 multimers, were here combined with laser capture microdissection to detect and collect SIV Gag CM9 specific CD8 + T cells in lymph node tissue from SIV infected rhesus macaques. CD8 + T cells from SIV infected and uninfected rhesus macaques were also collected and compared to the SIV GagCM9 specific CD8 + T cells. Illumina bead array and transcriptional analyses were used to assess the transcriptional profiles and the three different CD8 + T cell populations displayed unique transcriptional patterns. This pilot study demonstrates that rapid and specific immunostaining combined with laser capture microdissection in concert with transcriptional profiling may be used to elucidate phenotypic differences between CD8 + T cells in SIV infection. Such technologies may be useful to determine differences in functional activities of HIV/SIV specific T cells.

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          Temporal association of cellular immune responses with the initial control of viremia in primary human immunodeficiency virus type 1 syndrome.

          Virologic and immunologic studies were performed on five patients presenting with primary human immunodeficiency virus type 1 (HIV-1) infection. CD8+ cytotoxic T lymphocyte (CTL) precursors specific for cells expressing antigens of HIV-1 Gag, Pol, and Env were detected at or within 3 weeks of presentation in four of the five patients and were detected in all five patients by 3 to 6 months after presentation. The one patient with an absent initial CTL response had prolonged symptoms, persistent viremia, and low CD4+ T-cell count. Neutralizing antibody activity was absent at the time of presentation in all five patients. These findings suggest that cellular immunity is involved in the initial control of virus replication in primary HIV-1 infection and indicate a role for CTL in protective immunity to HIV-1 in vivo.
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            Control of viremia in simian immunodeficiency virus infection by CD8+ lymphocytes.

            Clinical evidence suggests that cellular immunity is involved in controlling human immunodeficiency virus-1 (HIV-1) replication. An animal model of acquired immune deficiency syndrome (AIDS), the simian immunodeficiency virus (SIV)-infected rhesus monkey, was used to show that virus replication is not controlled in monkeys depleted of CD8+ lymphocytes during primary SIV infection. Eliminating CD8+ lymphocytes from monkeys during chronic SIV infection resulted in a rapid and marked increase in viremia that was again suppressed coincident with the reappearance of SIV-specific CD8+ T cells. These results confirm the importance of cell-mediated immunity in controlling HIV-1 infection and support the exploration of vaccination approaches for preventing infection that will elicit these immune responses.
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              Dramatic Rise in Plasma Viremia after CD8+ T Cell Depletion in Simian Immunodeficiency Virus–infected Macaques

              To determine the role of CD8+ T cells in controlling simian immunodeficiency virus (SIV) replication in vivo, we examined the effect of depleting this cell population using an anti-CD8 monoclonal antibody, OKT8F. There was on average a 99.9% reduction of CD8 cells in peripheral blood in six infected Macaca mulatta treated with OKT8F. The apparent CD8 depletion started 1 h after antibody administration, and low CD8 levels were maintained until day 8. An increase in plasma viremia of one to three orders of magnitude was observed in five of the six macaques. The injection of a control antibody to an infected macaque did not induce a sustained viral load increase, nor did it significantly reduce the number of CD8+ T cells. These results demonstrate that CD8 cells play a crucial role in suppressing SIV replication in vivo.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                17 March 2016
                2016
                : 11
                : 3
                : e0149907
                Affiliations
                [1 ]Department of Medicine Solna, Unit of Infectious Diseases, Center for Molecular Medicine, Karolinska Institutet, Karolinska University Hospital, L8:01, 17176 Stockholm, Sweden
                [2 ]National Laboratory for HIV Immunology, Public Health Agency of Canada, Winnipeg, Manitoba, Canada
                [3 ]Department of Medical Microbiology, University of Manitoba, 730 William Ave. Winnipeg, MB, Canada
                [4 ]Department of Biosciences and Nutrition, Karolinska Institutet, Karolinska University Hospital Huddinge, Huddinge, Stockholm, Sweden
                [5 ]Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, United States of America
                [6 ]Department of Medicine, University of Washington, Seattle, WA, United States of America
                [7 ]Department of Laboratory Medicine, University of Washington, Seattle, WA, United States of America
                [8 ]Department of Pathology, Vaccine and Gene Therapy Institute, and the Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR, United States of America
                University of Pittsburgh, UNITED STATES
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: AT MJM LC. Performed the experiments: AT AB JL TP JZ LJP. Analyzed the data: AT AB JL TP JZ LÖ KB LC. Contributed reagents/materials/analysis tools: AB JL LJP. Wrote the paper: AT AB JL TP JZ LÖ LJP KB MJM LC.

                Article
                PONE-D-15-46881
                10.1371/journal.pone.0149907
                4795610
                26986062
                fb865642-f1c2-4ee7-9c67-5be9d847e425
                © 2016 Tjernlund et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 9 November 2015
                : 5 February 2016
                Page count
                Figures: 4, Tables: 3, Pages: 17
                Funding
                This work was supported by Swedish Research Council, http://www.vr.se/ (A.T.), the Swedish Society for Medical Research, http://www.ssmf.se/ (A.T.), the Swedish Society of Medicine, http://www.sls.se/ (A.T.), the Swedish Physicians Against AIDS Research Foundation http://www.aidsfond.se/ (A.T.), the Bill & Melinda Gates Foundation #41185 “Immune Correlates of Protection Against HIV and SIV Infection”, http://www.gatesfoundation.org/ (M.J.M.) and National Institutes of Health grant R01 AI-42528 and PO1 AI-30731, http://www.nih.gov/ (L.C.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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