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Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques
Published on: April 22, 2010
Antigen-specific cytokine responses in vaccinated Macaca nemestrina
T Mulvania1, J B Lynch, M N Robertson
1Department of Microbiology, University of Washington, Seattle, USA. mulvania@u.washington.edu
Insights
A novel assay distinguishes cytotoxic T lymphocyte (CTL) activity from suppressive cytokines. This HIV-2 vaccine study in macaques revealed that robust CTL responses correlate with protective T-helper type 1 activity.
Area of Science:
- Immunology
- Virology
- Vaccinology
Background:
- CD8+ T lymphocyte activity is crucial for controlling viral infections.
- Distinguishing cytotoxic T lymphocyte (CTL) activity from cytokine-mediated suppression is challenging.
- Human immunodeficiency virus type 2 (HIV-2) infection models are used to study vaccine efficacy.
Purpose of the Study:
- To develop and apply a new surrogate assay for measuring CD8+ T lymphocyte activity.
- To differentiate between CTL-mediated cytotoxicity and cytokine-driven suppression.
- To evaluate the immune response in macaques vaccinated with an attenuated HIV-2 strain.
Main Methods:
- Development of a novel surrogate assay for CD8+ T lymphocyte activity.
- Application of the assay to two groups of Macaca nemestrina vaccinated with HIV-2(KR).
- Group 1 received an additional inoculation with non-infectious HIV-2287, followed by challenge with infectious HIV-2287 for both groups.
Main Results:
- Five of six animals in Group 1 were protected against CD4 decline, compared to three of six in Group 2.
- Strong CTL responses against HIV-2(KR)-Gag were observed in Group 1.
- Antigen-specific T-helper (Th) type 1 responses correlated with strong CTL responses.
Conclusions:
- The new assay effectively distinguishes CTL activity from suppressive cytokine activity.
- Vaccination with an attenuated HIV-2 strain, followed by exposure to a pathogenic strain, elicits protective immune responses.
- Antigen-specific cytokine profiles, particularly Th1 responses, are associated with CD8+ T-cell-mediated protection against HIV-2.
Abstract:
We describe a new surrogate assay for CD8 + T lymphocyte activity that has the capability of discriminating between cytotoxic T lymphocyte (CTL) activity and cytokine-mediated suppressive activity. We applied this approach to two groups of Macaca nemestrina vaccinated with a minimally pathogenic strain of human immunodeficiency virus type 2 [HIV-2 (HIV-2(KR))] as a model of an attenuated virus vaccine. Group 1 was then inoculated with a non-infectious stock of a pathogenic strain, HIV-2287. Both groups 1 and 2 were subsequently challenged with an infectious stock of HIV-2287. Five out of six group 1 animals were protected against CD4 decline, whereas three out of six animals in group 2 were protected. Analysis of CTL responses demonstrated strong activity against HIV-2(KR)-Gag in group 1. It was determined that strong CTL responses correlate with antigen-specific T-helper (Th) type 1 responses. This antigen-specific cytokine assay has the potential to better elucidate the functional mechanisms of CD8 + T-cell-mediated protection than traditional methods to date.
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