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Updated: May 30, 2026
Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation
Published on: December 15, 2017
Quantitative assessment of macrophages in the muscularis externa of mouse intestines
H B Mikkelsen1, J O Larsen, P Froh
1Department of Cellular and Molecular Medicine, Faculty of Health Sciences, University of Copenhagen, Blegdamsvej 3, Copenhagen N, Denmark. hannem@sund.ku.dk
Insights
Quantifying intestinal CD169(+) and major histocompatibility class II antigen (MHCII(+)) cells in mice revealed that cell densities vary significantly by age and intestinal location. Young mice generally had higher cell densities than adults, with specific patterns observed in the jejunum, ileum, and colon.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- Quantifying intestinal cells is complex due to variations in density, cell morphology, and the need for precise immunolabeling.
- Accurate identification and quantification of specific cell populations, such as CD169(+) and MHCII(+) cells, are crucial for understanding intestinal immunity.
Purpose of the Study:
- To describe and validate a method for immunolabeling and quantifying CD169(+) and MHCII(+) cells in the mouse intestinal muscularis externa.
- To investigate the impact of age (young vs. adult mice) and anatomical location (jejunum, ileum, colon) on the densities of these cell populations.
Main Methods:
- Utilized design-based stereology for cell quantification in the intestinal muscularis externa of young (5-week-old) and adult (10-week-old) mice.
- Employed immunolabeling techniques to identify CD169(+) and major histocompatibility class II antigen (MHCII(+)) cells.
- Analyzed cell densities across different intestinal segments (jejunum, ileum, colon) and along the oral-anal axis.
Main Results:
- Cell densities were higher in the jejunum/ileum compared to the colon.
- In adults, jejunum/ileum cell densities increased orally to anally, while young mice showed peak densities mid-intestine.
- Colon cell densities decreased orally to anally in both age groups. Young animals generally exhibited higher densities, except for MHCII(+) cells in the colon. CD169 and MHCII appear co-expressed in the jejunum/ileum.
Conclusions:
- Intestinal CD169(+) and MHCII(+) cell densities are significantly influenced by both the age of the animal and the specific region of the intestine.
- The findings provide a quantitative basis for studying immune cell dynamics in the intestinal muscularis externa.
- Methodological details are provided for reproducible immunolabeling and stereological quantification of these critical immune cell populations.
Abstract:
Quantification of intestinal cells is challenging for several reasons: The cell densities vary throughout the intestines and may be age dependent. Some cell types are ramified and/or can change shape and size. Additionally, immunolabeling is needed for the correct identification of cell type. Immunolabeling is dependent on both up- and down-regulation of the antigen being labeled as well as on the primary and secondary antibodies, the fixation, and the enhancement procedures. Here, we provide a detailed description of immunolabeling of CD169(+) cells and major histocompatibility class II antigen (MHCII(+) ) cells and the subsequent quantification of these cells using design-based stereology in the intestinal muscularis externa. We used young (5-weeks-old) and adult (10-weeks-old) mice. Cell densities were higher in jejunum-ileum, when compared with colon. In jejunum/ileum, the cell densities increased in oral-anal direction in adults, whereas the densities were highest in the midpart in young animals. In colon, the cell densities decreased in oral-anal direction in both groups of animals. Except for the density of MHCII(+) cells in colon, the cell densities were highest in young animals. Densities of CD169(+) and MHCII(+) cells did not differ, except in the colon of young animals where the CD169(+) density was almost twice as high as the MHCII(+) density. CD169 and MHCII antigens seem to be expressed simultaneously by the same cell in jejunum/ileum. We conclude that cell densities depend on both the age of the mouse and on the location in the intestines.
