Document Detail


A fluorescence-based assay for quantitation of lymphokine-activated killer cell activity.
MedLine Citation:
PMID:  2785144     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A fluorescence assay for the quantitation of tumor cell lysis by activated and non-activated killer (LAK) cells is described. The target cells are labelled with a europium chelate (Eu-diethylenetriaminopentaacetate) and after cytolysis caused by the LAK cells the Eu3+ complex is released into the culture supernatant. The addition of beta-naphthoyltrifluoroacetone to culture supernatant aliquots leads to the formation of a highly fluorescent chelate which can be measured with a time-resolved fluorometer. The influence of various assay parameters has been evaluated including incubation time, effector-to-target cell ratio, the target cell line and different concentrations of interleukin-2 during cell culture. The optimized time-resolved fluorometric assay was found to be as simple and sensitive as the commonly used cytotoxicity assay in which the release of 51Cr from the labelled target cells is measured. In addition the assay is much faster and safer since the label is not radioactive.
Authors:
T Volgmann; A Klein-Struckmeier; H Mohr
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of immunological methods     Volume:  119     ISSN:  0022-1759     ISO Abbreviation:  J. Immunol. Methods     Publication Date:  1989 Apr 
Date Detail:
Created Date:  1989-06-02     Completed Date:  1989-06-02     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  1305440     Medline TA:  J Immunol Methods     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  45-51     Citation Subset:  IM    
Affiliation:
Blood Transfusion Service of Lower Saxony, Department of Research and Development, Springe, F.R.G.
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MeSH Terms
Descriptor/Qualifier:
Cell Communication
Chelating Agents
Chromium Radioisotopes / metabolism
Cytotoxicity Tests, Immunologic* / methods
Europium / metabolism
Fluoroimmunoassay / methods*
Humans
Interleukin-2*
Killer Cells, Natural / enzymology,  immunology*,  metabolism
L-Lactate Dehydrogenase / metabolism
Lymphocyte Activation*
Time Factors
Chemical
Reg. No./Substance:
0/Chelating Agents; 0/Chromium Radioisotopes; 0/Interleukin-2; 7440-53-1/Europium; EC 1.1.1.27/L-Lactate Dehydrogenase

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


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