Spatial and temporal control of T cell activation using a photoactivatable agonist Journal Article
| Authors: | Sanchez, E.; Huse, M. |
| Article Title: | Spatial and temporal control of T cell activation using a photoactivatable agonist |
| Abstract: | T lymphocytes engage in rapid, polarized signaling, occurring within minutes following TCR activation. This induces formation of the immunological synapse, a stereotyped cell-cell junction that regulates T cell activation and directionally targets effector responses. To study these processes effectively, an imaging approach that is tailored to capturing fast, polarized responses is necessary. This protocol describes such a system, which is based on a photoactivatable peptide-major histocompatibility complex (pMHC) that is non-stimulatory until it is exposed to ultraviolet light. Targeted decaging of this reagent during videomicroscopy experiments enables precise spatiotemporal control of TCR activation and high-resolution monitoring of subsequent cellular responses by total internal reflection (TIRF) imaging. This approach is also compatible with genetic and pharmacological perturbation strategies. This allows for the assembly of well-defined molecular pathways that link TCR signaling to the formation of the polarized cytoskeletal structures that underlie the immunological synapse. © 2018 Journal of Visualized Experiments. |
| Keywords: | signal transduction; immunology; cell polarity; t lymphocytes; centrosome; photochemistry; cell biology; t cell receptor (tcr); immunology and infection; issue 134; major histocompatibility complex (pmhc); total internal reflection microscopy (tirf) |
| Journal Title: | Journal of Visualized Experiments |
| Volume: | 2018 |
| Issue: | 134 |
| ISSN: | 1940-087X |
| Publisher: | MYJoVE Corporation |
| Date Published: | 2018-04-01 |
| Start Page: | e56655 |
| Language: | English |
| DOI: | 10.3791/56655 |
| PROVIDER: | scopus |
| PUBMED: | 29757266 |
| PMCID: | PMC6100773 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 1 June 2018 -- Source: Scopus |
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