Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

Hong He; Amy N. Courtney; Eric Wieder; K. Jagannadha Sastry

doi:10.3791/1743

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

JoVE Journal
Immunology and Infection

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JoVE Journal Immunology and Infection

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

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07:11 min

April 22, 2010

DOI: 10.3791/1743-v

Hong He¹, Amy N. Courtney¹, Eric Wieder², K. Jagannadha Sastry¹

¹Department of Immunology,MD Anderson Cancer Center - University of Texas, ²Department of Medicine,University of Miami

We demonstrate the utility of multicolor flow cytometry for detailed phenotypic and functional characterization of total as well as memory subsets of CD4+ and CD8+ T cells in rhesus macaques, the ideal model for HIV/AIDS vaccine studies.

This video demonstrates a procedure for stimulating and staining peripheral blood mononuclear cells or pbmc. For multicolor flow cytometry analysis, Alec Watts of pbmc isolated from whole blood are placed in tissue culture plates. Cells are chemically or biologically stimulated to produce an immune response.

Following stimulation, a cocktail of antibodies is added to identify cell types based on expressed surface markers. The cells are then permeated and a different cocktail of antibodies is added to detect the expression of intracellular cytokines, cells are then analyzed by facts to determine the cytokine profiles for specific subsets of pbmc. Hi, I'm Amy Courtney from Dr.Jagan SAS's lab in the Department of Immunology at the University of Texas MB Anderson Cancer Center.

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