# Immunology Core

> **NIH NIH P01** · UNIVERSITY OF MARYLAND BALTIMORE · 2024 · $413,764

## Abstract

IMMUNOLOGY CORE – ABSTRACT
Diarrheal disease is the second leading cause of death in children under five years of age. Understanding the
processes involved in bacterial pathogenesis and host response to enteric pathogens is essential to improve
treatment and develop effective prevention strategies. Such studies have been hindered by the lack of reliable
models that fully recapitulate the complex cellular and molecular events and interactions that take place in the
human gut. The Immunology Core (IC) will establish and refine co-culture models of human intestinal epithelial
and innate immune cells to recreate the tissue structure, cell interaction, and coordinated responses of the
human gut. This physiologically relevant system will generate new insights into the coordinated actions of
epithelial and immune cells during enteric infections. The IC will also support the studies performed by the
projects by providing technical expertise for implementation of immune co-cultures. The IC has two main Aims.
Aim 1. As a core developmental Aim, the IC will: 1) Refine the enteroid-immune cell co-culture by seeding
epithelial cell monolayers on a collagen-coated porous scaffold (i.e. Alvetex®) that will allow direct cell-to-cell
contact and migration. 2) Establish a multi-cellular model consisting of epithelial cell monolayers co-cultured with
different innate phagocytic cells: monocyte-derived macrophages (MF), polymorphonuclear neutrophils (PMN),
and dendritic cells (DC) isolated from human peripheral blood. Phagocytic cells not only perform anti-microbial
functions but also initiate adaptive immunity. We have successfully engrafted human phagocytes individually to
enteroid monolayers. To better recreate individual and synergistic function of epithelial and innate immune cells,
we will combine these three main phagocytic lineages with enteroid monolayers. Tissue structure and integration,
cell phenotype, viability, and functionality will be assessed by histology, confocal microscopy, flow cytometry and
microarray technology. Phagocytic activity, transmigration and cytokine and chemokine production will be
examined to determine cell function. 3) Develop a co-culture model containing epithelial cells and TCRgd+ T cells
from human peripheral blood. TCRgd+ T cells within the mucosal barrier, also known as intraepithelial
lymphocytes (IEL), are rapidly mobilized and deploy diverse innate immune functions in response to pathogens.
Cell phenotype will be characterized by flow cytometry, and function will be assessed by cytokine production and
cell cytotoxicity. Tissue structure will be examined by histology and microscopy. Cell movement will be monitored
with particle tracking system/software and confocal microscopy. Optimal conditions for the generation of immune
co-cultures will be defined, and protocols produced and made available to the research projects.
Aim 2. As a support service Aim, the IC will write detailed protocols to implement the methods described ...

## Key facts

- **NIH application ID:** 10868680
- **Project number:** 5P01AI125181-09
- **Recipient organization:** UNIVERSITY OF MARYLAND BALTIMORE
- **Principal Investigator:** Marcela F Pasetti
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $413,764
- **Award type:** 5
- **Project period:** 2016-07-01 → 2026-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10868680

## Citation

> US National Institutes of Health, RePORTER application 10868680, Immunology Core (5P01AI125181-09). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10868680. Licensed CC0.

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