A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling
Inmaculada Vico-Barranco , Mikel M. Arbulo-Echevarria , Serrano-García , Alba Pérez-Linaza , José M. Miranda-Sayago , Arkadiusz Miążek , Isaac Narbona-Sánchez , Enrique Aguado
AbstractIntracellular signaling through the T cell receptor (TCR) is essential for T cell development and function. Proper TCR signaling requires the sequential activities of Lck and ZAP-70 kinases, which result in the phosphorylation of tyrosine residues located in the CD3 ITAMs and the LAT adaptor, respectively. LAT, linker for the activation of T cells, is a transmembrane adaptor protein that acts as a scaffold coupling the early signals coming from the TCR with downstream signaling pathways leading to cellular responses. The leukemic T cell line Jurkat and its derivative mutants J.CaM1.6 (Lck deficient) and J.CaM2 (LAT deficient) have been widely used to study the first signaling events upon TCR triggering. In this work, we describe the loss of LAT adaptor expression found in a subline of J.CaM1.6 cells and analyze cis-elements responsible for the LAT expression defect. This new cell subline, which we have called J.CaM1.7, can re-express LAT adaptor after Protein Kinase C (PKC) activation, which suggests that activation-induced LAT expression is not affected in this new cell subline. Contrary to J.CaM1.6 cells, re-expression of Lck in J.CaM1.7 cells was not sufficient to recover TCR-associated signals, and both LAT and Lck had to be introduced to recover activatory intracellular signals triggered after CD3 crosslinking. Overall, our work shows that the new LAT negative J.CaM1.7 cell subline could represent a new model to study the functions of the tyrosine kinase Lck and the LAT adaptor in TCR signaling, and their mutual interaction, which seems to constitute an essential early signaling event associated with the TCR/CD3 complex.
|Journal series||Cells, ISSN 2073-4409, e-ISSN 2073-4409, (N/A 140 pkt)|
|Publication size in sheets||0.95|
|Keywords in English||Lck; LAT; J.CaM1.6; TCR; signaling|
|License||Journal (articles only); published final; ; with publication|
|Score||= 140.0, 04-04-2021, ArticleFromJournal|
|Publication indicators||= 0; : 2017 = 1.344; : 2017 = 4.829 (2)|
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