Abstract
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-U1 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 (M(r)) 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.
| Original language | English |
|---|---|
| Pages (from-to) | 253-259 |
| Number of pages | 7 |
| Journal | International Journal of Cancer |
| Volume | 36 |
| Issue number | 2 |
| Publication status | Published - 1985 |
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ASJC Scopus subject areas
- Cancer Research
- Oncology
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Selection and characterization of monoclonal antibodies to the idiotype-like structure of an interleukin-2-producing human leukemia T-cell line. / Moretta, A.; Pantaleo, G.; Lopez-Botet, M.; Moretta, L.
In: International Journal of Cancer, Vol. 36, No. 2, 1985, p. 253-259.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Selection and characterization of monoclonal antibodies to the idiotype-like structure of an interleukin-2-producing human leukemia T-cell line
AU - Moretta, A.
AU - Pantaleo, G.
AU - Lopez-Botet, M.
AU - Moretta, L.
PY - 1985
Y1 - 1985
N2 - 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-U1 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 (M(r)) 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.
AB - 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-U1 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 (M(r)) 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.
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M3 - Article
C2 - 2410377
AN - SCOPUS:0022416667
VL - 36
SP - 253
EP - 259
JO - International Journal of Cancer
JF - International Journal of Cancer
SN - 0020-7136
IS - 2
ER -