Document Detail


ELISpot assay as a tool to study oxidative stress in peripheral blood mononuclear cells.
MedLine Citation:
PMID:  21057930     Owner:  NLM     Status:  In-Process    
Abstract/OtherAbstract:
Enzyme-Linked Immuno Spot (ELISpot) assay is widely used for vaccine development, cancer and AIDS research, and autoimmune disease studies. The output of an ELISpot assay is a formation of colored spots which appear at the sites of cells releasing cytokines, with each individual spot representing a single cytokine-releasing cell.We have shown that hydrogen peroxide-induced oxidative stress was causing ∼twofold decrease in the number of lymphocytes secreting the TH1 cytokines IFN-gamma and IL-2, as well as chemokine IL-8 and cytokine TNF alpha. However, the number of cells secreting TH2 cytokines IL-4 and IL-5 in hydrogen peroxide-treated group did not change. Our ELISpot data indicate that oxidative stress may affect TH1-TH2 cytokine secretion balance which, in turn, may underlie developments of various pathological conditions. We adopted ELISpot assay for studying oxidative stress in human peripheral blood lymphocytes by analyzing the acute effect of hydrogen peroxide treatment on the frequency of cells secreting IFN-gamma, IL-2, IL-4, IL-5, IL-8, and TNF-alpha.
Authors:
Jodi Hagen; Jeffrey P Houchins; Alexander E Kalyuzhny
Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  682     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2011  
Date Detail:
Created Date:  2010-11-08     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  207-15     Citation Subset:  IM    
Affiliation:
R&D Systems, Inc, Minneapolis, MN, USA.
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