Document Detail


Measurement of T cell activation after 16-hr in vitro stimulation with concanavalin A.
MedLine Citation:
PMID:  20069526     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A flow cytometry assay that can be used to directly determine the proportion of activated T lymphocytes in human whole blood samples after stimulation with concanavalin A is presented here. Human whole blood is incubated with fluorescently labeled antibodies (against CD3, CD4, CD8, and CD69), erythrocytes are then lysed, and the samples are analyzed using a flow cytometer. The assay presented is able to differentiate between CD4+ and CD8+ T lymphocytes. Thus, it is possible to quantify both lymphocyte populations in parallel, as well as the respective proportions of activated T lymphocytes, all from one sample. An additional advantage of this assay is that it was developed to assay whole blood, thereby reducing the likelihood of introducing pre-analytic error and facilitating the calculation of quantitative ratios. Thus, the method can be used for both in vitro and ex vivo experiments.
Authors:
Borros M Arneth
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Current protocols in cytometry / editorial board, J. Paul Robinson, managing editor ... [et al.]     Volume:  Chapter 6     ISSN:  1934-9300     ISO Abbreviation:  Curr Protoc Cytom     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-13     Completed Date:  2010-03-09     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100899351     Medline TA:  Curr Protoc Cytom     Country:  United States    
Other Details:
Languages:  eng     Pagination:  Unit 6.28.1-10     Citation Subset:  IM    
Affiliation:
Institute of Clinical Chemistry and Laboratory Medicine, Johannes Gutenberg University Mainz, Mainz, Germany.
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal / pharmacology
CD4-Positive T-Lymphocytes / drug effects,  immunology
CD8-Positive T-Lymphocytes / drug effects,  immunology
Cell Extracts
Concanavalin A / immunology*,  pharmacology
Erythrocytes / drug effects
Flow Cytometry / methods*
Humans
Immunophenotyping
Lymphocyte Activation* / drug effects
Lymphocyte Subsets / drug effects
Time Factors
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Cell Extracts; 11028-71-0/Concanavalin A

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


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