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| Item Type: | Article |
|---|---|
| Title: | Gauging antigen recognition by human primary T-cells featuring orthotopically exchanged TCRs of choice |
| Creators Name: | Mühlgrabner, V., Plach, A., Holler, J., Leitner, J., Steinberger, P., Dupré, L., Göhring, J. and Huppa, J.B. |
| Abstract: | Understanding human T-cell antigen recognition in health and disease is becoming increasingly instrumental for monitoring T-cell responses to pathogen challenge and for the rational design of T-cell-based therapies targeting cancer, autoimmunity and organ transplant rejection. Here we showcase a quantitative imaging platform which is based on the use of planar glass-supported lipid bilayers (SLBs). The latter are functionalized with antigen (peptide-loaded HLA) as adhesion and costimulatory molecules (ICAM-1, B7-1) to serve as surrogate antigen presenting cell for antigen recognition by T-cells, which are equipped with T-cell antigen receptors (TCRs) sequenced from antigen-specific patient T-cells. We outline in detail, how the experimental use of SLBs supports recoding and analysis of synaptic antigen engagement and calcium signaling at the single cell level in response to user-defined antigen densities for quantitative comparison. |
| Keywords: | TCR Sequencing, T-Cell Antigen Sensitivity, CRISPR-Cas9-Mediated TCR Exchange, Advanced Live-Cell Imaging |
| Source: | Methods in Cell Biology |
| Series Name: | Methods in Cell Biology |
| Title of Book: | The Immunological Synapse - Part C |
| ISSN: | 0091-679X |
| ISBN: | 978-0-443-21868-2 |
| Publisher: | Elsevier |
| Volume: | 193 |
| Page Range: | 127-154 |
| Number of Pages: | 264 |
| Date: | 5 February 2025 |
| Official Publication: | https://doi.org/10.1016/bs.mcb.2024.03.003 |
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