Linked Life Data

11121557

Source:http://linkedlifedata.com/resource/pubmed/id/11121557

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
1-2
pubmed:dateCreated
2001-1-2
pubmed:abstractText
The detection of antigen specific clonal T-cell populations in vivo during T-cell selection and an immune responses is often hampered due to the lack of suitable clonotype specific monoclonal antibodies. In order to determine the potential usefulness of green fluorescent protein (GFP) to follow specific T-cells in vivo, we decided to express and analyze the function of a T-cell receptor (TCR) alpha chain-GFP fusionprotein. The TCRalpha and beta chain cDNAs of a Leishmania major-specific murine T helper 2 cell clone were cloned and inserted into the pHSE3' expression vector. Simultaneously, a TCRalpha expression vector was constructed containing a C-terminal in frame fusion with the open reading frame of the enhanced GFP (EGFP). TCRalpha/TCRbeta or TCRalpha-EGFP/TCRbeta constructs were expressed in T-cell hybridoma cells 58alpha(-)beta(-) which lack an endogenous TCR but still express CD3 components. The TCRalpha-EGFP fusionprotein was detected with the expected molecular weight by immunoprecipitation and Western Blot analysis. Surface staining of TCR components was detected in transfectants expressing the wild type TCR heterodimer and, with only a slight reduction in intensity, also in those expressing the TCR-EGFP complex. Hence, expression and transport to the outer cell membrane is possible despite the 27 kD C-terminal extension of the TCRalpha. Most importantly, the EGFP-tagged TCR was functional since the transfectants produced IL-2 in response to stimulation via their TCR. Thus, TCR-EGFP constructs represent attractive tools to study posttranslational regulation of TCR expression and ligand-induced TCR clustering as well as the fate of antigen specific T-cells during tolerance induction and immunity in transgenic mouse models.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0022-1759
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
246
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
165-74
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:11121557-Animals, pubmed-meshheading:11121557-Biological Markers, pubmed-meshheading:11121557-Cell Membrane, pubmed-meshheading:11121557-Clone Cells, pubmed-meshheading:11121557-Epitopes, T-Lymphocyte, pubmed-meshheading:11121557-Green Fluorescent Proteins, pubmed-meshheading:11121557-Humans, pubmed-meshheading:11121557-Luminescent Proteins, pubmed-meshheading:11121557-Mice, pubmed-meshheading:11121557-Mice, Inbred BALB C, pubmed-meshheading:11121557-Receptors, Antigen, T-Cell, alpha-beta, pubmed-meshheading:11121557-Recombinant Fusion Proteins, pubmed-meshheading:11121557-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:11121557-Signal Transduction, pubmed-meshheading:11121557-T-Lymphocytes, pubmed-meshheading:11121557-Th2 Cells, pubmed-meshheading:11121557-Transfection
pubmed:year
2000
pubmed:articleTitle
A stable marker for specific T-cells: a TCR alpha/green fluorescent protein (GFP) fusionprotein reconstitutes a functionally active TCR complex.
pubmed:affiliation
Clinical Microbiology, Immunology and Hygiene, University of Erlangen-Nürnberg, Wasserturmstrasse 3, 91054, Erlangen, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't