Detection of cell surface ligands for human synovial γδ T cells
| Autor: | Robin A. Ross, Heather Brouwer, Yuan Lui, Bryan A Ballif, Ralph C. Budd, Anisha Mahalya Gogerly-Moragoda, Ana Filipa L.O.M. Santos, Kuberan Balagurunathan, Sumana Sharma, Gavin J. Wright, Simon J. Davis, Cheryl Collins |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
T cell
Immunology Cell Ligands Lymphocyte Activation Monocytes Flow cytometry Cell Line 03 medical and health sciences 0302 clinical medicine Antigen Downregulation and upregulation T-Lymphocyte Subsets medicine Immunology and Allergy Humans Receptor medicine.diagnostic_test Ligand Chemistry Synovial Membrane Receptors Antigen T-Cell gamma-delta 3. Good health Cell biology medicine.anatomical_structure Cell culture Clinical and Human Immunology Protein Multimerization Glycolysis 030215 immunology |
| Zdroj: | The Journal of Immunology Author Choice |
| ISSN: | 1550-6606 |
| DOI: | 10.4049/jimmunol.1900451 |
| Popis: | Key Points TCR-γδ tetramer identified ligand expression by flow cytometry. TCR-γδ ligands were induced on activated monocytes or T cells. Bioinformatics combined with mass spectrometry produced an overlapping list of 16 candidate ligands. Visual Abstract Lack of understanding of the nature and physiological regulation of γδ T cell ligands has considerably hampered full understanding of the function of these cells. We developed an unbiased approach to identify human γδ T cells ligands by the production of a soluble TCR-γδ (sTCR-γδ) tetramer from a synovial Vδ1 γδ T cell clone from a Lyme arthritis patient. The sTCR-γδ was used in flow cytometry to initially define the spectrum of ligand expression by both human tumor cell lines and certain human primary cells. Analysis of diverse tumor cell lines revealed high ligand expression on several of epithelial or fibroblast origin, whereas those of hematopoietic origin were largely devoid of ligand. This allowed a bioinformatics-based identification of candidate ligands using RNAseq data from each tumor line. We further observed that whereas fresh monocytes and T cells expressed low to negligible levels of TCR-γδ ligands, activation of these cells resulted in upregulation of surface ligand expression. Ligand upregulation on monocytes was partly dependent upon IL-1β. The sTCR-γδ tetramer was then used to bind candidate ligands from lysates of activated monocytes and analyzed by mass spectrometry. Surface TCR-γδ ligand was eliminated by treatment with trypsin or removal of glycosaminoglycans, and also suppressed by inhibition of endoplasmic reticulum–Golgi transport. Of particular interest was that inhibition of glycolysis also blocked TCR-γδ ligand expression. These findings demonstrate the spectrum of ligand(s) expression for human synovial Vδ1 γδ T cells as well as the physiology that regulates their expression. |
| Databáze: | OpenAIRE |
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