γδ intraepithelial lymphocytes facilitate pathological epithelial cell shedding via CD103-mediated granzyme release

Hu, Madeleine D., Golovchenko, Natasha B., Burns, Grace L., Nair, Prema M., Kelly, Thomas J., Agos, Jonathan, Zand Irani, Mudar, Sinn Soh, Wai, Zeglinski, Matthew R., Lemenze, Alexander, Bonder, Edward M., Sandrock, Inga, Prinz, Immo, Granville, David J., Keely, Simon, Watson, Alastair J. M. and Edelblum, Karen L. (2022) γδ intraepithelial lymphocytes facilitate pathological epithelial cell shedding via CD103-mediated granzyme release. Gastroenterology, 162 (3). 877-889.e7. ISSN 0016-5085

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Abstract

Background & Aims: Excessive shedding of apoptotic enterocytes into the intestinal lumen is observed in inflammatory bowel disease and is correlated with disease relapse. Based on their cytolytic capacity and surveillance behavior, we investigated whether intraepithelial lymphocytes expressing the γδ T cell receptor (γδ IELs) are actively involved in the shedding of enterocytes into the lumen. Methods: Intravital microscopy was performed on GFP γδ T cell reporter mice treated with intraperitoneal lipopolysaccharide (10 mg/kg) for 90 minutes to induce tumor necrosis factor–mediated apoptosis. Cell shedding in various knockout or transgenic mice in the presence or absence of blocking antibody was quantified by immunostaining for ZO-1 funnels and cleaved caspase-3 (CC3). Granzyme A and granzyme B release from ex vivo–stimulated γδ IELs was quantified by enzyme-linked immunosorbent assay. Immunostaining for γδ T cell receptor and CC3 was performed on duodenal and ileal biopsies from controls and patients with Crohn's disease. Results: Intravital microscopy of lipopolysaccharide-treated mice revealed that γδ IELs make extended contact with shedding enterocytes. These prolonged interactions require CD103 engagement by E-cadherin, and CD103 knockout or blockade significantly reduced lipopolysaccharide-induced shedding. Furthermore, we found that granzymes A and B, but not perforin, are required for cell shedding. These extracellular granzymes are released by γδ IELs both constitutively and after CD103/E-cadherin ligation. Moreover, we found that the frequency of γδ IEL localization to CC3-positive enterocytes is increased in Crohn's disease biopsies compared with healthy controls. Conclusions: Our results uncover a previously unrecognized role for γδ IELs in facilitating tumor necrosis factor–mediated shedding of apoptotic enterocytes via CD103-mediated extracellular granzyme release.

Item Type: Article
Additional Information: Funding Information: Funding Cell sorting was performed at the Rutgers New Jersey Medical School Flow Cytometry and Immunology Core Laboratory and was supported by National Institute for Research Resources grant S10RR027022. This work was supported by National Institute of Health grants R21AI143892 (K.L.E), T32AI125185 , F30DK121391 (M.D.H.) and the New Jersey Health Foundation (K.L.E.), grants BB/J004529/1 and BB/K018256/1 (A.J.M.W.), and National Health and Medical Research Council Project grant and CRE Digestive Health (S.K.). D.J.G. was funded by a Canadian Institutes for Health Research Foundation Grant.
Uncontrolled Keywords: cell extrusion,gamma delta t cells,mucosal immunology,hepatology,gastroenterology ,/dk/atira/pure/subjectarea/asjc/2700/2721
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
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Depositing User: LivePure Connector
Date Deposited: 08 Dec 2021 09:32
Last Modified: 01 Mar 2022 04:30
URI: https://ueaeprints.uea.ac.uk/id/eprint/82596
DOI: 10.1053/j.gastro.2021.11.028

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