Production of lacZ inducible T cell hybridoma specific for human and mouse gp10025-33 peptides
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1
Weizmann Institute of Science, Israel
The key event in T cell activation is the recognition of a peptide bound to major histocompatibility complex (MHC) molecules on the surface of antigen presenting cells (APCs). The enormous pool of peptides displayed on MHC makes it almost impossible to detect a given peptide-MHC complex on the surface of APCs by using conventional indirect methods. On the other hand, direct recognition of a selected peptide by the TCR results in generation of intracellular signals leading to initiation of the primary stages of T cell activation. To facilitate measurement of T cell activation and to enable identification of individual clones, β-galactosidase (lacZ) inducible CD4+ and CD8+ T cell hybrids were developed. Previous studies showed that heterologous Escherichia coli β-galactosidase (lacZ) gene, under control of the IL-2 entire enhancer region or the nuclear factor in activated T cells (NFAT) element alone, is specifically induced in transfected and activated T cells. Thus, activation of transfected T cells, results in synthesis of both IL-2 and lacZ gene products. Moreover, since the lacZ remains sequestered within the activated cells, chromogenic or fluorogenic substrate enables measurement of an activating event in a single T cell. Generation of the hybrids is relatively easy and allows maintenance in culture and the lacZ assay provides a rapid, sensitive and non-radioactive method for measuring T cell activation. In this study we isolated T cells from Pmel-1 mice and generated a lacZ inducible CD8+ T cell hybridoma.
Keywords:
hybridoma,
tcr,
gp100 Melanoma Antigen,
Antigen Presentation,
activation
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Immune receptors and signaling
Citation:
Cafri
G,
Sharbi-Yunger
A,
Tzehoval
E,
Alteber
Z,
Azulay
M,
Gozlan
YM and
Eisenbach
L
(2013). Production of lacZ inducible T cell hybridoma specific for human and mouse gp10025-33 peptides.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00586
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Received:
05 Jun 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Prof. Lea Eisenbach, Weizmann Institute of Science, Rehovot, Israel, lea.eisenbach@weizmann.ac.il