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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
3
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pubmed:dateCreated |
1983-9-20
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pubmed:abstractText |
A method allowing quantitative analysis of the expression of foreign antigens at the surface of transformed cells is described. Aminoethyl-Sephadex G-25 beads labelled with different amounts of fluorescein isothiocyanate (FITC) were used to calibrate an Epics V cell sorter. The quantity of FITC molecules bound per bead was found to be a linear function of relative fluorescence intensity expressed by channel number for intermediate and high levels of fluorescence and a non-linear function for low levels. The lowest limit of detectable fluorescence was 3400 FITC molecules bound per bead. Using FITC-conjugated monoclonal antibodies (m.Ab.) the number of HLA class I molecules expressed at the surface of murine LMTK- (H-2Kk) cells transfected by cloned HLA class I genes was estimated and compared with the amount expressed on human B (Raji) and T (MOLT 4) lymphoblastoid reference cells. Murine transformed cells expressed approximately 3 times less HLA class I determinants per surface unit (micrometer 2) than Raji cells and 1.4 times less than MOLT 4 cells. Similar results were obtained by Scatchard analysis of the same populations using radiolabelled m.Ab. A detailed analysis of fluorescent cells showed that 5-20% of transformed cells expressed less than 33,000 HLA class I molecules/cell whereas most MOLT 4 cells exhibited at least 280,000 molecules/cell and the majority of Raji cells more than 700,000 molecules/cell. The expression of foreign antigen did not reduce the amount of H-2Kk molecules at the surface of transformed cells (mean of 350,000 molecules/cell).
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescein-5-isothiocyanate,
http://linkedlifedata.com/resource/pubmed/chemical/Fluoresceins,
http://linkedlifedata.com/resource/pubmed/chemical/Fluorescent Dyes,
http://linkedlifedata.com/resource/pubmed/chemical/HLA Antigens,
http://linkedlifedata.com/resource/pubmed/chemical/Thiocyanates,
http://linkedlifedata.com/resource/pubmed/chemical/Thymidine Kinase
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pubmed:status |
MEDLINE
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pubmed:month |
Jul
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pubmed:issn |
0022-1759
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
29
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pubmed:volume |
61
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
301-15
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:6409961-Animals,
pubmed-meshheading:6409961-Burkitt Lymphoma,
pubmed-meshheading:6409961-Cell Line,
pubmed-meshheading:6409961-Cloning, Molecular,
pubmed-meshheading:6409961-Flow Cytometry,
pubmed-meshheading:6409961-Fluorescein-5-isothiocyanate,
pubmed-meshheading:6409961-Fluoresceins,
pubmed-meshheading:6409961-Fluorescent Dyes,
pubmed-meshheading:6409961-HLA Antigens,
pubmed-meshheading:6409961-Humans,
pubmed-meshheading:6409961-L Cells (Cell Line),
pubmed-meshheading:6409961-Major Histocompatibility Complex,
pubmed-meshheading:6409961-Mice,
pubmed-meshheading:6409961-Thiocyanates,
pubmed-meshheading:6409961-Thymidine Kinase
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pubmed:year |
1983
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pubmed:articleTitle |
Quantification by flow cytofluorimetry of HLA class I molecules at the surface of murine cells transformed by cloned HLA genes.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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