Reconstitution of an active surface CD2 by DNA transfer in CD2-CD3+ jurkat cells facilitates CD3-T cell receptor-mediated IL-2 production

Hafedh Makni, James S. Malter, John C. Reed, Sano Nobuhiko, Georgina Lang, Dimitris Kioussis, Giorgio Trinchieri, Malek Kamoun

Research output: Contribution to journalArticle

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Abstract

To investigate the requirements for CD2 expression in the activation of T lymphocytes via the CD3-TCR complex, we produced and characterized a series of CD2-variants of the IL-2 producing Jurkat leukemia cell line, J32 (surface phenotype, CD2+, CD3+, CD28+). These mutants were derived by radiation and immunoselection, and were cloned under limiting dilution conditions. A total of 3 out of 30 of these mutants selectively lost the expression of both CD2 surface molecules and CD2 mRNA, and retained the expression of the CD3-TCR complex and the CD28 molecule. A mitogenic combination of anti-CD2 antibodies (9.6 + 9-1) failed to stimulate activation of these variants as measured by mobilization of intracellular Ca2+ and by IL-2 production. The CD2- mutants stimulated with anti-CD3 or anti-TCR mAb revealed an 8- to 32-fold decrease in IL-2 production and IL-2 mRNA accumulation as compared with the parental cells. No alteration of CD3-TCR-induced mobilization of intracellular Ca2+ was observed in the CD2- mutants. Reconstitution of CD2 expression by gene transfer in two J32 CD2- mutants restored IL-2 production and IL-2 mRNA accumulation in responses to both anti-CD2 and anti-CD3-TCR mAb. These results are the first direct demonstration of the requirement for CD2 molecules in optimizing IL-2 response in human T cells stimulated via CD3-TCR complex.

Original languageEnglish (US)
Pages (from-to)2522-2529
Number of pages8
JournalJournal of Immunology
Volume146
Issue number8
StatePublished - Apr 15 1991

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Jurkat Cells
T-Cell Antigen Receptor
Interleukin-2
T-Cell Antigen Receptor-CD3 Complex
DNA
Messenger RNA
T-Lymphocytes
Anti-Idiotypic Antibodies
Leukemia
Radiation
Phenotype
Gene Expression
Cell Line

ASJC Scopus subject areas

  • Immunology

Cite this

Reconstitution of an active surface CD2 by DNA transfer in CD2-CD3+ jurkat cells facilitates CD3-T cell receptor-mediated IL-2 production. / Makni, Hafedh; Malter, James S.; Reed, John C.; Nobuhiko, Sano; Lang, Georgina; Kioussis, Dimitris; Trinchieri, Giorgio; Kamoun, Malek.

In: Journal of Immunology, Vol. 146, No. 8, 15.04.1991, p. 2522-2529.

Research output: Contribution to journalArticle

Makni, H, Malter, JS, Reed, JC, Nobuhiko, S, Lang, G, Kioussis, D, Trinchieri, G & Kamoun, M 1991, 'Reconstitution of an active surface CD2 by DNA transfer in CD2-CD3+ jurkat cells facilitates CD3-T cell receptor-mediated IL-2 production', Journal of Immunology, vol. 146, no. 8, pp. 2522-2529.
Makni, Hafedh ; Malter, James S. ; Reed, John C. ; Nobuhiko, Sano ; Lang, Georgina ; Kioussis, Dimitris ; Trinchieri, Giorgio ; Kamoun, Malek. / Reconstitution of an active surface CD2 by DNA transfer in CD2-CD3+ jurkat cells facilitates CD3-T cell receptor-mediated IL-2 production. In: Journal of Immunology. 1991 ; Vol. 146, No. 8. pp. 2522-2529.
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