Selection and characterization of monoclonal antibodies to the idiotype-like structure of an interleukin-2-producing human leukemia T-cell line

Détails

ID Serval
serval:BIB_06D3EADC50D6
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Selection and characterization of monoclonal antibodies to the idiotype-like structure of an interleukin-2-producing human leukemia T-cell line
Périodique
International Journal of Cancer
Auteur(s)
Moretta  A., Pantaleo  G., Lopez-Botet  M., Moretta  L.
ISSN
0020-7136 (Print)
Statut éditorial
Publié
Date de publication
08/1985
Volume
36
Numéro
2
Pages
253-9
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Aug 15
Résumé
A variant of the Jurkat leukemia cell line termed JA3 (surface phenotype: T11+., T3+, T4-, T8-, T6-, T1+, 3A1+, HLA-DR-, Tac-, 4F2+) has been used for mouse immunization and production of monoclonal antibodies (MAbs) to molecules carrying clonotypic determinants that are thought to serve as receptor for antigen on human T cells. JA3 had been selected because of its ability to release large amounts of IL-2 following stimulation with phytohemagglutinin (PHA) or anti-T3 antibody in the presence of phorbolmyristate acetate (PMA). This functional property has been exploited for screening of MAbs potentially directed to idiotype-like structures of JA3 cells. Thus supernatants of hybridomas (obtained by fusing mouse splenocytes with P3-UI myeloma cells) were analyzed for their ability to induce JA3 cells to release IL-2 in the presence of PMA. Four stimulatory antibodies reacted with JA3 but not with polyclonal T-cell populations, T-cell clones, or T and B tumor cell lines as assessed by indirect immunofluorescence and fluorescence-activated cell sorter (FACS) analysis. All 4 antibodies immunoprecipitated the same disulphide-linked heterodimeric molecules having a molecular mass (Mr) of approximately 85,000 under non-reducing conditions that was resolved in 2 major peptides of 40,000 and 45,000 under reducing conditions. These data indicate that these antibodies (termed anti-JTi) were directed to the clonotypic-restricted structures of the JA3 T-cell-receptor molecules. Unlike the anticlonotypic antibodies described so far, anti-JTi MAbs were capable of triggering IL-2 production even in unbound, soluble form, in the absence of adherent cells or PMA. Competitive inhibition experiments, in which 35S-labelled anti-JTi MAbs have been used, provided evidence that they may be directed against different epitopes on the same clonotypic structure.
Mots-clé
Antibodies, Monoclonal/biosynthesis/*immunology Antigens, Neoplasm/immunology Antigens, Surface/immunology Cell Line Epitopes/immunology Flow Cytometry Fluorescent Antibody Technique Humans Immunochemistry Immunoglobulin Idiotypes/*immunology Interleukin-2/biosynthesis Leukemia, Lymphocytic/*blood Precipitation Receptors, Antigen, T-Cell/*immunology T-Lymphocytes/*immunology/metabolism
Pubmed
Web of science
Création de la notice
25/01/2008 16:14
Dernière modification de la notice
03/03/2018 13:26
Données d'usage