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Article

A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling

1
Institute of Biomedical Research Cadiz (INIBICA), 11009 Cádiz, Spain
2
Rheumatology Section, Unit of Orthopedic Surgery, Traumatology and Rheumatology, HUPM, 11009 Cádiz, Spain
3
Department of Biochemistry and Molecular Biology, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland
4
Department of Biomedicine, Biotechnology and Public Health (Immunology), Universidad de Cádiz, 11002 Cádiz, Spain
*
Author to whom correspondence should be addressed.
Academic Editors: Gerhard J. Schütz and Johannes Huppa
Cells 2021, 10(2), 343; https://doi.org/10.3390/cells10020343
Received: 17 December 2020 / Revised: 2 February 2021 / Accepted: 3 February 2021 / Published: 6 February 2021
(This article belongs to the Special Issue Molecular Mechanisms of Early T Cell Signaling)
Intracellular 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. View Full-Text
Keywords: Lck; LAT; J.CaM1.6; TCR; signaling Lck; LAT; J.CaM1.6; TCR; signaling
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MDPI and ACS Style

Vico-Barranco, I.; Arbulo-Echevarria, M.M.; Serrano-García, I.; Pérez-Linaza, A.; Miranda-Sayago, J.M.; Miazek, A.; Narbona-Sánchez, I.; Aguado, E. A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling. Cells 2021, 10, 343. https://doi.org/10.3390/cells10020343

AMA Style

Vico-Barranco I, Arbulo-Echevarria MM, Serrano-García I, Pérez-Linaza A, Miranda-Sayago JM, Miazek A, Narbona-Sánchez I, Aguado E. A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling. Cells. 2021; 10(2):343. https://doi.org/10.3390/cells10020343

Chicago/Turabian Style

Vico-Barranco, Inmaculada, Mikel M. Arbulo-Echevarria, Isabel Serrano-García, Alba Pérez-Linaza, José M. Miranda-Sayago, Arkadiusz Miazek, Isaac Narbona-Sánchez, and Enrique Aguado. 2021. "A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR Signaling" Cells 10, no. 2: 343. https://doi.org/10.3390/cells10020343

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