A Novel, LAT/Lck Double Deficient T Cell Subline J.CaM1.7 for Combined Analysis of Early TCR 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.This research was funded by Consejeria de Salud de Andalucia, Junta de Andalucia (grant PI-0055-2017 to E.A.), and Fundacion Biomedica Cadiz Proyectos INIBICA 2019 (grant LI19/I14NCO15 to E.A. and M.M.A.-E.)

Expression of Non-T Cell Activation Linker (NTAL) in Jurkat Cells Negatively Regulates TCR Signaling: Potential Role in Rheumatoid Arthritis

T lymphocytes are key players in adaptive immune responses through the recognition of peptide antigens through the T Cell Receptor (TCR). After TCR engagement, a signaling cascade is activated, leading to T cell activation, proliferation, and differentiation into effector cells. Delicate control of activation signals coupled to the TCR is needed to avoid uncontrolled immune responses involving T cells. It has been previously shown that mice deficient in the expression of the adaptor NTAL (Non-T cell activation linker), a molecule structurally and evolutionarily related to the transmembrane adaptor LAT (Linker for the Activation of T cells), develop an autoimmune syndrome characterized by the presence of autoantibodies and enlarged spleens. In the present work we intended to deepen investigation into the negative regulatory functions of the NTAL adaptor in T cells and its potential relationship with autoimmune disorders. For this purpose, in this work we used Jurkat cells as a T cell model, and we lentivirally transfected them to express the NTAL adaptor in order to analyze the effect on intracellular signals associated with the TCR. In addition, we analyzed the expression of NTAL in primary CD4+ T cells from healthy donors and Rheumatoid Arthritis (RA) patients. Our results showed that NTAL expression in Jurkat cells decreased calcium fluxes and PLC-γ1 activation upon stimulation through the TCR complex. Moreover, we showed that NTAL was also expressed in activated human CD4+ T cells, and that the increase of its expression was reduced in CD4+ T cells from RA patients. Our results, together with previous reports, suggest a relevant role for the NTAL adaptor as a negative regulator of early intracellular TCR signaling, with a potential implication in RA.14 página

Mortality in emergency services

El envejecimiento progresivo de la población parece ser uno de los hechos relacionados con el incremento en la demanda de servicios sanitarios. Los servicios de urgencia hospitalarios actúan muchas veces como medio de canalización de los pacientes que acuden a ellos en busca de atención médica. No obstante, estos servicios pueden encontrarse con frecuencia al borde de la saturación debido al elevado número de personas que acuden a sus puertas. Las patologías atendidas por los médicos de urgencias son muy diferentes y de diversa gravedad. Una proporción variable de los pacientes que son atendidos en urgencias fallece en los mismos. Durante el año 2014, se realizó un estudio en el servicio de urgencias del Hospital “Marqués de Valdecilla” con el objetivo de conocer el tipo de paciente que fallece en el mismo, así como las causas más frecuentes de mortalidad. Se observó, como ya se venía describiendo en la literatura, que los pacientes de edad avanzada y con patología crónica son los que suelen fallecer en urgencias. Así mismo, patologías respiratorias, de carácter infeccioso y cardiovasculares son las principales protagonistas como causas de mortalidad en este servicio de urgencias.The progressive aging of the population seems to be one of the main facts in relation with the increased demand in health services. The hospital emergency services usually act like a canalization way for those patients who come to them in order to get health attention. However, these services may meet at the edge of the saturation due to the high number of patients who come to their doors. The pathologies that doctors take care of in emergency services are very different and diverse gravity. A variable proportion of patients who are attended in these emergency services die there. During 2014, a study was made in the emergency service of the Hospital “Marqués de Valdecilla” in order to know the profile patient who usually die there and, at the same time, the most frequent causes of mortality. As the literature reflects, aged patients with chronic pathologies usually die in emergency services. In the same way, respiratory, infectious and cardiovascular pathologies are the principal subjects as dead causes in emergency services.Grado en Medicin