Source:http://linkedlifedata.com/resource/pubmed/id/17222422
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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
1-2
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pubmed:dateCreated |
2007-3-12
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pubmed:abstractText |
The single color IFN-gamma ELISPOT assay has become a standard for assessing HIV-specific immune responses in HIV-infected subjects. However, recent data suggests that single cytokine detection for immune monitoring of HIV-infected individuals may not be sufficient to fully describe virus-specific immune responses. Here, we have designed and validated a dual color ELISPOT assay capable of detecting both IL-2 and IFN-gamma secreting cells simultaneously in response to HIV antigens. We found that a cell input number of 200,000 cells/well provided a good balance between limited availability of cells due to blood volume restrictions and ability to detect all cytokine secretion patterns. The simultaneous detection of IL-2 and IFN-gamma resulted in a decreased magnitude of IFN-gamma but not IL-2 responses. Measures of intra- and inter-assay variability for the dual color ELISPOT assay were comparable to that seen for single cytokine ELISPOT assay with coefficients of variation below 20% for IL-2, IFN-gamma and dual secretion. Although CD8+ T cells mediated most HIV-specific responses in infected subjects, CD4+ T cells mediated responses to HIV were also detected. Features of this assay such as high throughput, cell number requirement and cytokine choice should make this assay a valuable tool for screening for HIV-specific immune responses in several clinically relevant settings.
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pubmed:grant | |
pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Mar
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pubmed:issn |
0022-1759
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
30
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pubmed:volume |
320
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
18-29
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pubmed:dateRevised |
2011-10-17
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pubmed:meshHeading |
pubmed-meshheading:17222422-Cells, Cultured,
pubmed-meshheading:17222422-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:17222422-HIV Infections,
pubmed-meshheading:17222422-Humans,
pubmed-meshheading:17222422-Interferon-gamma,
pubmed-meshheading:17222422-Interleukin-2,
pubmed-meshheading:17222422-Peptides,
pubmed-meshheading:17222422-T-Lymphocytes
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pubmed:year |
2007
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pubmed:articleTitle |
A dual color ELISPOT method for the simultaneous detection of IL-2 and IFN-gamma HIV-specific immune responses.
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pubmed:affiliation |
Research Institute of the McGill University Health Center, Montreal, Canada.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't,
Evaluation Studies,
Research Support, N.I.H., Extramural
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