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Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes

Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes

Published on: May 8, 2016

Microparticles and their impact on intestinal immunity.

Helen M Becker¹, Martina M Bertschinger, Gerhard Rogler

¹Department of Medicine, University of Calgary, Calgary, Alta., Canada.

Digestive Diseases (Basel, Switzerland)

|January 9, 2013

Summary

Titanium dioxide (TiO2) microparticles, common in food, are ingested by immune cells and may activate the NLRP3 inflammasome, potentially worsening gastrointestinal diseases like Crohn's disease.

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Area of Science:

Immunology
Gastroenterology
Nanotechnology

Background:

Microparticles, including titanium dioxide (TiO2), are prevalent in the Western diet as food additives.
Their accumulation in the gut, particularly in Peyer's patches, raises concerns about potential links to gastrointestinal diseases like Crohn's disease (CD).
The NLRP3 inflammasome, a key regulator of inflammation, is activated by certain particles, suggesting TiO2 as a potential target for study.

Purpose of the Study:

To investigate the uptake and effects of titanium dioxide (TiO2) microparticles on immune and intestinal cells.
To determine if TiO2 can activate the NLRP3 inflammasome pathway.
To explore the potential role of TiO2 in exacerbating gastrointestinal inflammation.

Main Methods:

Incubation of macrophage-like cells and intestinal epithelial cells with TiO2 microparticles.
Assessment of TiO2 uptake by cells using microscopy and cell counting.
Analysis of NLRP3 inflammasome assembly and activation, including caspase-1 activity.
Measurement of proinflammatory cytokine secretion, specifically interleukin (IL)-1β and IL-18.

Main Results:

Macrophage-like cells readily ingested TiO2 microparticles within 6 hours.
TiO2 exposure led to the assembly of NLRP3 with caspase-1, indicating inflammasome activation.
This inflammasome assembly correlated with increased secretion of IL-1β.
Intestinal epithelial cells also ingested TiO2 in a dose-dependent manner.
TiO2 particles were found intracellularly in both cell types.

Conclusions:

TiO2 microparticles are ingested by intestinal epithelial cells and macrophages.
TiO2 can activate the NLRP3 inflammasome, leading to IL-1β and IL-18 secretion.
In individuals with compromised intestinal barriers (e.g., IBD patients), TiO2 uptake and subsequent inflammasome activation may aggravate existing gastrointestinal inflammation.