7 results match your criteria: "Universidad Complutense de Madrid and 12 de Octubre Health Research Institute (Imas12)[Affiliation]"

Introduction: CD8 cytotoxic T lymphocytes (CTLs) are highly effective in defending against viral infections and tumours. They are activated through the recognition of peptide-MHC-I complex by the T-cell receptor (TCR) and co-stimulation. This cognate interaction promotes the organisation of intimate cell-cell connections that involve cytoskeleton rearrangement to enable effector function and clearance of the target cell.

View Article and Find Full Text PDF
Article Synopsis
  • Understanding T cell function requires exploring the molecular dynamics involved in T cell activation and effector function.
  • The Jurkat (JK) CD4 T cell line serves as a valuable in vitro model for studying the intracellular signaling pathways triggered by T cell receptor (TCR) activation.
  • The text outlines protocols to assess TCR and costimulation-mediated activation, immunological synapse formation, and cytokine production, which are key processes in T cell responses across various pathophysiological contexts.
View Article and Find Full Text PDF

Activation kinetics of regulatory molecules during immunological synapse in T cells.

Methods Cell Biol

July 2023

Immunology Unit from Hospital Universitario de la Princesa, Universidad Autónoma de Madrid and Instituto de investigación Sanitaria La Princesa (IIS-IP), Madrid, Spain; Centro de Investigación Biomédica en Red Enfermedades Cardiovasculares (CIBERCV), Madrid, Spain. Electronic address:

T cell activation through TCR stimulation leads to the formation of the immunological synapse (IS), a specialized adhesion organized between T lymphocytes and antigen presenting cells (APCs) in which a dynamic interaction among signaling molecules, the cytoskeleton and intracellular organelles achieves proper antigen-mediated stimulation and effector function. The kinetics of molecular reactions at the IS is essential to determine the quality of the response to the antigen stimulation. Herein, we describe methods based on biochemistry, flow cytometry and imaging in live and fixed cells to study the activation state and dynamics of regulatory molecules at the IS in the Jurkat T cell line CH7C17 and primary human and mouse CD4 T lymphocytes stimulated by antigen presented by Raji and HOM2 B cell lines and human and mouse dendritic cells.

View Article and Find Full Text PDF

T cell-redirecting strategies have emerged as effective cancer immunotherapy approaches. Bispecific antibodies (bsAbs) are designed to specifically recruit T cells to the tumor microenvironment and induce the assembly of the immunological synapse (IS) between T cells and cancer cells or antigen-presenting cells. The way that the quality of the IS might predict the effectiveness of T cell-redirecting strategies, including those mediated by bsAbs or by chimeric antigen receptors (CAR)-T cells, is currently under discussion.

View Article and Find Full Text PDF

Spatial and temporal regulation of molecular reactions dictates cell fate. Thus, studying molecular dynamics is essential to understand how cells decide what to do and the fundamental perturbations causing disease. Classically, molecular dynamics has been studied by protocols based in the overexpression of fluorescent fusion proteins.

View Article and Find Full Text PDF

It has been proposed that the accumulation of farnesylated phosphatase of regenerating liver-1 (PRL-1) at the plasma membrane is mediated by static electrostatic interactions of a polybasic region with acidic membrane lipids and assisted by oligomerization. Nonetheless, localization at early and recycling endosomes suggests that the recycling compartment might also contribute to its plasma membrane accumulation. Here, we investigated in live cells the dynamics of PRL-1 fused to the green fluorescent protein (GFP-PRL-1).

View Article and Find Full Text PDF