Cryptosporidium infection of human small intestinal epithelial cells induces type III interferon and impairs infectivity of Rotavirus

ABSTRACT

Cryptosporidiosis is a major cause of severe diarrheal disease in infants from resource poor settings. The majority of infections are caused by the human-specific pathogen C. hominis and absence of in vitro growth platforms has limited our understanding of host-pathogen interactions and development of effective treatments. To address this problem, we developed a stem cell-derived culture system for C. hominis using human enterocytes differentiated under air-liquid interface (ALI) conditions. Human ALI cultures supported robust growth and complete development of C. hominis in vitro including all life cycle stages. Cryptosporidium infection induced a strong interferon response from enterocytes, possibly driven, in part, by an endogenous dsRNA virus in the parasite. Prior infection with Cryptosporidium induced type III IFN secretion and consequently blunted infection with Rotavirus, including live attenuated vaccine strains. The development of hALI provides a platform for further studies on human-specific pathogens, including clinically important coinfections that may alter vaccine efficacy.

KEYWORDS:

• Vaccine
• mucosal immunity
• interferon
• antiviral
• co-infection
• enteric infection
• parasitology

Acknowledgments

We thank Soumya Ravindran for initial studies and Kelli VanDussen and Yi Wang for helpful advice on developing the culture methods used here. We would like to acknowledge Matthew Freese for his technical support. Select illustrations were kindly provided by Abigail Kimball in association with InPrint at Washington University in St. Louis. Sequencing was performed by the Genome Technology Access Core, McDonnell Genome Institute, Washington University. Imaging was performed using the Molecular Microbiology Imaging Facility, Washington University School of Medicine. Histological services and spheroid cell lines were provided by the Digestive Diseases Research Core Center (NIDDK P30 DK052574), Washington University School of Medicine. Rotarix (RV1) and Rotateq (RV5) stocks were a kind gift from Kristen Ogden at Vanderbilt University. Immunology assays were conducted with support of the Bursky Center for Human Immunology and Immunotherapy Programs at Washington University, Immuno-monitoring Laboratory. A preprint of an earlier version of this manuscript has been posted to BioRxiv doi: https://doi.org/10.1101/2023.08.30.555581.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Conceptualization: Valentin Greigert, Siyuan Ding, L. David Sibley. Data curation: Valentin Greigert, Iti Saraav, Juhee Son, Yinxing Zhu. Formal Analysis: Valentin Greigert, Iti Saraav. Investigation: Valentin Greigert, Iti Saraav, Juhee Son, Yinxing Zhu. Visualization: Valentin Greigert. Writing – original draft: Valentin Greigert. Resources: Denise Dayao, Avan Antia, Saul Tzipori, William H. Witola, Thaddeus S. Stappenbeck. Supervision: Siyuan Ding, Saul Tzipori, L. David Sibley. Writing – review & editing: Valentin Greigert, Siyuan Ding, L. David Sibley.

Data availability statement

Raw RNA-seq reads and analyzed data generated in this study have been deposited in the Gene Expression Omnibus database under accession numbers GSE241570 (C. hominis) & GSE241571 (C. parvum). All raw data and R scripts, as well as supplementary table 1, are available on github.com/Sibley-Lab/Chominis-hALI.git.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/19490976.2023.2297897

Additional information

Funding

Support for studies in the Sibley laboratory was provided by agrant from Open Philanthropy [GV673603895]. Support for studies conducted in the Ding lab were provided by NIH T32 DK007130, U19 AI116484, and R01 AI150796.