Investigating the role of CD47 in γδ intraepithelial lymphocyte migration

Intraepithelial lymphocytes expressing the γδ T cell receptor (γδ IELs) survey the intestinal epithelial barrier to limit enteric pathogen translocation. Type I interferon (IFN) is key mediator of innate immunity and upregulates the expression of CD47 (integrin associated protein), a ubiquitously expressed transmembrane protein involved in neutrophil transepithelial migration. Based on this, we hypothesized that type I IFN induces CD47 to enhance γδ IEL migration into the lateral intercellular space (LIS). To test this, γδ IEL/enteroid co-cultures were treated with type I IFN resulting in a 23% increase in γδ IEL migration into the LIS compared to untreated controls (p<0.05). Similar findings were observed in vivo as intravital microscopy revealed a 30% increase in γδ IEL track speed following administration of type I IFN (p<0.0001). Transcriptomic analysis of sorted γδ IELs isolated from IFNα-treated mice showed increased expression of cell surface associated IFN-stimulated genes, Cd47 and Jaml (3.5 and 4.7-fold, respectively) relative to PBS controls (p<0.05). Interestingly, treatment of γδ IEL/enteroid co-cultures with anti-CD47, but not anti-JAML, abrogated the type I IFN-induced increase in γδ IEL migration. To determine if loss of epithelial or γδ T cell CD47 expression affects IEL subsets or total number of γδ IELs, we crossed γδ T cell reporter mice to those with global or conditional deletion of CD47. Loss of epithelial CD47 was verified by immunofluorescence. Morphometric analysis showed no effect on total jejunal γδ IEL number in any of the CD47-deficient mice, and flow cytometry revealed that CD47 is not required to maintain individual IEL subsets or basal integrin expression on γδ IELs. Moreover, intravital microscopy reveals that mice lacking epithelial CD47 showed a 14% increase in γδ IEL migratory speed at steady-state compared to littermate controls (p<0.0001), reflecting a trend of reduced dwell time in the LIS. Thus, we posit that epithelial CD47 may play a role in regulating the length of time a γδ IEL interacts with individual enterocytes. Additional studies to address the role of γδ T cell CD47 on basal and type I IFN-mediated γδ IEL migratory behavior are ongoing. This work was supported by NIH R01DK119349. This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.