Limit-dilution assay and clonal expansion of all T cells capable of proliferation.

Détails

ID Serval
serval:BIB_519E9EB6D1A1
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Limit-dilution assay and clonal expansion of all T cells capable of proliferation.
Périodique
Journal of Immunological Methods
Auteur(s)
Chen W.F., Wilson A., Scollay R., Shortman K.
ISSN
0022-1759 (Print)
ISSN-L
0022-1759
Statut éditorial
Publié
Date de publication
1982
Volume
52
Numéro
3
Pages
307-322
Langue
anglais
Résumé
A limit-dilution microculture system is presented in which almost all mature T cells, cultured at a level of about 1 cell/well, grow and expand to clones averaging 60,000 cells over an 8-9 day period. Cloning efficiency is 70-100%, so the set of the expanded clones is representative of the starting T-cell population. T cells of all Lyt phenotypes form clones of progeny cells. The system involves culture in flat-bottom microtitre trays, in the presence of concanavalin A as the initiating stimulus, together with appropriately irradiated spleen filler cells and a supplementary source of soluble T cell growth factors. The resultant clones may be screened for cytolytic function, as described in the accompanying paper. The system may be used to assay the level of T cells capable of expansion or precursor function (PTL-p) by using [3H]TdR uptake as a readout for the presence or absence of proliferating clones. Analysis of the frequency of positive cultures shows a good fit to the expected Poisson distribution, with no evidence of complicating suppressor or helper effects.
Mots-clé
Animals, Antigens, Ly/genetics, Antilymphocyte Serum/pharmacology, Clone Cells/cytology, Clone Cells/immunology, Concanavalin A/pharmacology, Cytotoxicity Tests, Immunologic/methods, Interleukin-2/biosynthesis, Kinetics, Lymphocyte Activation/radiation effects, Mice, Spleen/cytology, Spleen/immunology, T-Lymphocytes/classification, T-Lymphocytes/cytology
Pubmed
Web of science
Création de la notice
10/04/2013 11:21
Dernière modification de la notice
03/03/2018 17:11
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