Document Detail


Multicolor flow cytometry analyses of cellular immune response in rhesus macaques.
MedLine Citation:
PMID:  20414189     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The rhesus macaque model is currently the best available model for HIV-AIDS with respect to understanding the pathogenesis as well as for the development of vaccines and therapeutics(1,2,3). Here, we describe a method for the detailed phenotypic and functional analyses of cellular immune responses, specifically intracellular cytokine production by CD4+ and CD8+ T cells as well as the individual memory subsets. We obtained precise quantitative and qualitative measures for the production of interferon gamma (INF-) and interleukin (IL) -2 in both CD4+ and CD8+ T cells from the rhesus macaque PBMC stimulated with PMA plus ionomycin (PMA+I). The cytokine profiles were different in the different subsets of memory cells. Furthermore, this protocol provided us the sensitivity to demonstrate even minor fractions of antigen specific CD4+ and CD8+ T cell subsets within the PBMC samples from rhesus macaques immunized with an HIV envelope peptide cocktail vaccine developed in our laboratory. The multicolor flow cytometry technique is a powerful tool to precisely identify different populations of T cells (4,5) with cytokine-producing capability(6) following non-specific or antigen-specific stimulation (5,7).
Authors:
Hong He; Amy N Courtney; Eric Wieder; K Jagannadha Sastry
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Video-Audio Media     Date:  2010-04-22
Journal Detail:
Title:  Journal of visualized experiments : JoVE     Volume:  -     ISSN:  1940-087X     ISO Abbreviation:  J Vis Exp     Publication Date:  2010  
Date Detail:
Created Date:  2010-04-23     Completed Date:  2010-04-30     Revised Date:  2012-04-23    
Medline Journal Info:
Nlm Unique ID:  101313252     Medline TA:  J Vis Exp     Country:  United States    
Other Details:
Languages:  eng     Pagination:  -     Citation Subset:  IM    
Affiliation:
Department of Immunology, The University of Texas M.D. Anderson Cancer Center, TX, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
CD4-Positive T-Lymphocytes / immunology
CD8-Positive T-Lymphocytes / immunology
Disease Models, Animal
Flow Cytometry / methods*
Immunity, Cellular
Interferon-gamma / biosynthesis
Interleukin-2 / biosynthesis
Leukocytes, Mononuclear / immunology
Macaca mulatta / blood,  immunology*
T-Lymphocyte Subsets / immunology
Grant Support
ID/Acronym/Agency:
AI46969/AI/NIAID NIH HHS; CA 16672/CA/NCI NIH HHS
Chemical
Reg. No./Substance:
0/Interleukin-2; 82115-62-6/Interferon-gamma

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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