# Development of a human intestinal microphysiological system for the study of immune responses to protozoan parasites

> **NIH NIH R01** · UNIVERSITY OF WISCONSIN-MADISON · 2024 · $769,091

## Abstract

PROJECT SUMMARY
Microphysiological systems have great potential for modeling human disease but advanced in vitro models of
parasitic infection and immunity are severely underrepresented. Parasites are major causes of morbidity and
mortality globally, infecting millions of people every year, yet, there are no effective vaccines available for any
enteric parasitic infection. Oral transmission via contaminated food or water is the most common route of
parasitic infection for humans, but our knowledge of parasite/host interactions, including how parasites interact
with immune cells to either cause disease or elicit a protective immune response, within the intestinal tract is
very limited. There is a critical need to create improved in vitro models of human immune-parasite interactions
to capture key features present during parasitic infection establishment and disease progression. To address
this need we will develop a microphysiological gut vasculature lumen system based on the LumeNEXT
microfluidic device system. This 3-dimensional cell culture device recapitulates the gut architecture and
includes a human intestinal epithelial lumen flanked by blood and lymphatic vasculature. With these advanced
in vitro models, we will introduce parasites into the intestinal epithelium and human immune cells into the
vasculature to monitor parasitic disease and immune response. Our goal is to create a microphysiological
system that can be used for the study of any protozoan parasite. For this proposal, we will use the protozoan
parasites Toxoplasma gondii (T. gondii) and Entamoeba histolytica (E. histolytica) because 1) they are human
pathogens of global importance, 2) they are on distinct branches of the protist evolutionarily tree, 3) they have
defined lab growth conditions and genetically tagged marker strains, 4) our lab has recently developed the
unique ability to produce large numbers of the highly infectious oocysts and cysts forms of both T. gondii. Our
data shows that T. gondii infection of the intestinal epithelial lumen in our in vitro model system elicits an active
immune response and migration of human immune cells from the vasculature. In this project, we will use these
3D biomimetic gut-vasculature lumen models to address critical knowledge gaps of the human immune
responses to T. gondii and E. histolytica. We will incorporate an anaerobic environment so that the immune
responses can be defined under hypoxic conditions. These experiments will provide the foundational
understanding of the human innate immune responses to intestinal T. gondii infection that are essential for
vaccine development. We will also model E. histolytica invasive disease using nutrient limitation and co-culture
with Clostridiodes difficile. The advances we will achieve in this proposal will allow the microbiology and
immunology fields to determine the immune responses to the biologically relevant stages of intestinal parasites
in human models.

## Key facts

- **NIH application ID:** 10881985
- **Project number:** 5R01AI172874-02
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** David J Beebe
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $769,091
- **Award type:** 5
- **Project period:** 2023-07-05 → 2028-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10881985

## Citation

> US National Institutes of Health, RePORTER application 10881985, Development of a human intestinal microphysiological system for the study of immune responses to protozoan parasites (5R01AI172874-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10881985. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*