title: Studying Immunoreceptor Signaling in Human T Cells Using Electroporation of In Vitro Transcribed mRNA.
abstract: The recognition bestowed upon T lymphocytes as key mediators of cellular immunity has been further attested by recent successful clinical studies using genetically modified T cells. With an ever-growing interest in the application of T cells to treat human malignancies, studying the molecular mechanisms of T cell activation, signaling, and function has become imperative. This, therefore, calls for the development of new easy-to-use and accurate models to investigate the biological phenomena that begin at the synaptic levels of T cell and antigen interactions to the ultimate exhaustion and death of the T cell. Here, we describe an approach to transiently express a chimeric molecule on the cell surface that permits activation and expansion of T cells, thereby providing a model to study T cell signaling.
authors.affiliation: Center for Cellular Immunotherapies and Department of Pathology and Laboratory Medicine Perelman School of Medicine, University of Pennsylvania, SPE 8-101 South Tower, 3400 Civic Center Blvd, Philadelphia, PA, 19104, USA.
authors.forename: Omkar
authors.initials: O
authors.lastname: Kawalekar
authors.affiliation: Center for Cellular Immunotherapies and Department of Pathology and Laboratory Medicine Perelman School of Medicine, University of Pennsylvania, SPE 8-101 South Tower, 3400 Civic Center Blvd, Philadelphia, PA, 19104, USA.
authors.forename: Carl H
authors.initials: CH
authors.lastname: June
authors.affiliation: Center for Cellular Immunotherapies and Department of Pathology and Laboratory Medicine Perelman School of Medicine, University of Pennsylvania, SPE 8-101 South Tower, 3400 Civic Center Blvd, Philadelphia, PA, 19104, USA. Milone@upenn.edu.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Milone
mesh_terms: D018274:Electroporation / Q000379:methods*; D006801:Humans; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins
keywords: Antigen; Costimulation; Immunotherapy; Receptors; Synthetic biology