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Author Spotlight: Investigating the Effects of Compounds on Intestinal Tissue Using 3D Human Cell Line Models

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Breaking Barriers: A Protocol to Investigate Intestinal Pathogen Adhesion, Invasion, and Translocation Through In

Rishi Drolia^1,2,3,4, Arun K Bhunia^5,6,7

¹Cellular and Molecular Microbiology Laboratory, Department of Biological Science, Old Dominion University, Norfolk, VA, USA. rdrolia@odu.edu.

Methods in Molecular Biology (Clifton, N.J.)

|June 11, 2025

Summary

This study explores how pathogens infect the gastrointestinal tract using combined in vitro and in vivo models. Advanced imaging reveals how microbes breach intestinal defenses, aiding in developing new infection therapies.

Keywords:

Caco-2 Cell adhesion Confocal microscopy Fluorescence in situ hybridization (FISH)Host–pathogen interactions Immunostaining Infection dynamics Intestinal epithelial barrier Intestinal epithelial cells Invasion Microbial pathogenesis Mouse and Gerbil models Tight junction proteins Translocation Z-stack imaging

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Area of Science:

Microbiology
Infectious Diseases
Cell Biology

Background:

The gastrointestinal tract is a key site for microbial pathogenesis.
Pathogen adhesion, invasion, and translocation are critical for infection establishment.
Understanding these host-pathogen interactions is vital for developing effective treatments.

Purpose of the Study:

To present a comprehensive approach for studying microbial pathogenesis in the gastrointestinal tract.
To elucidate host-pathogen interactions and infection dynamics at the cellular level.
To integrate in vitro and in vivo models with advanced imaging techniques.

Main Methods:

Utilized in vitro assays with intestinal cell models for adhesion, invasion, and translocation analysis.
Employed in vivo rodent models to study complex host-pathogen-tissue interactions.
Applied advanced imaging techniques, including confocal microscopy, immunostaining, and fluorescence in situ hybridization (FISH).

Main Results:

Demonstrated the ability to precisely analyze microbial interactions at the cellular level using in vitro assays.
Gained insights into host immune responses and tissue dynamics through in vivo models.
Visualized high-resolution host-pathogen interactions and microbial dissemination using advanced imaging.

Conclusions:

An integrated approach combining functional assays and advanced imaging enhances understanding of microbial pathogenesis.
This methodology reveals critical mechanisms of pathogen invasion and systemic spread.
The findings facilitate the development of novel therapeutic strategies against gastrointestinal infections.