Analysis of intestinal epithelial cell responses to Cryptosporidium highlights the temporal effects of IFN-gamma on parasite restriction

Pardy, R. D.; Walzer, K. A.; Wallbank, B. A.; Byerly, J. H.; O'Dea, K. M.; Cohn, I. S.; Haskins, B. E.; Roncaioli, J. L.; Smith, E. J.; Buenconsejo, G. Y.; Striepen, B.; Hunter, C. A.

doi:10.1101/2023.11.14.567008

bioRxiv · 10.1101/2023.11.14.567008

Analysis of intestinal epithelial cell responses to Cryptosporidium highlights the temporal effects of IFN-gamma on parasite restriction

Pardy, R. D.·Walzer, K. A.·Wallbank, B. A.·Byerly, J. H.·O'Dea, K. M.·Cohn, I. S.·Haskins, B. E.·Roncaioli, J. L.·Smith, E. J.·Buenconsejo, G. Y.·Striepen, B.·Hunter, C. A.

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Abstract

The production of IFN-{gamma} is crucial for control of multiple enteric infections, but its impact on intestinal epithelial cells (IEC) is not well understood. Cryptosporidium parasites exclusively infect epithelial cells and the ability of interferons to activate the transcription factor STAT1 in IEC is required for parasite clearance. The use of single cell RNA sequencing to profile IEC during infection revealed induction of IFN-{gamma}-dependent gene signatures that was comparable between uninfected and infected cells, and IEC expression of the IFN-{gamma} receptor was required for parasite control. Unexpectedly, treatment of Ifng-/- mice with IFN-{gamma} demonstrated the IEC response to this cytokine correlates with a delayed reduction in parasite burden but did not affect parasite development. These data sets provide insight into the impact of IFN-{gamma} on IEC and suggest a model in which IFN-{gamma}-mediated bystander activation of uninfected enterocytes is important for control of Cryptosporidium. AUTHOR SUMMARYThe cytokine interferon-gamma (IFN-{gamma}) plays an important role in the control of intracellular infections by a wide variety of bacteria, viruses and parasites. While the impact of IFN-{gamma} on immune cells has been a major research focus, how it impacts intestinal epithelial cells remains poorly understood. Cryptosporidium parasites are an important cause of morbidity in a variety of epidemiological settings and exclusively infect intestinal epithelial cells (IEC). Recent advances in the ability to genetically modify and study Cryptosporidium in wild-type hosts provides a useful model to investigate IEC-intrinsic mechanisms of pathogen control. In this study, single cell RNA-sequencing was used to analyze the IEC response to infection and IFN-{gamma} signalling. We demonstrate broad changes in the epithelial compartment during infection that include the induction of an IEC population with robust induction of IFN-{gamma}-stimulated genes. In addition, we show that infected IEC remain responsive to IFN-{gamma} signalling, and that this cytokine causes a delayed reduction in parasite burden that correlates with the kinetics of IEC responsiveness to IFN-{gamma} stimulation. Together, our work uncovers how Cryptosporidium infection impacts the IEC compartment and helps define the relationship between the kinetics of IFN-{gamma} responsiveness and pathogen control in IEC.

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Pardy, R. D., Walzer, K. A., Wallbank, B. A., Byerly, J. H., O'Dea, K. M., Cohn, I. S., Haskins, B. E., Roncaioli, J. L., Smith, E. J., Buenconsejo, G. Y., Striepen, B., Hunter, C. A.. 2023-11-16. Analysis of intestinal epithelial cell responses to Cryptosporidium highlights the temporal effects of IFN-gamma on parasite restriction. https://doi.org/10.1101/2023.11.14.567008

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