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MNPmApp: An image analysis tool to quantify mononuclear phagocyte distribution in mucosal tissues
Catherine Potts1, Julia Schearer2, Thomas A Sebrell2
1Department of Mathematical Sciences, Montana State University, Bozeman, Montana, USA.
Abstract:
Mononuclear phagocytes (MNPs) such as dendritic cells and macrophages perform key sentinel functions in mucosal tissues and are responsible for inducing and maintaining adaptive immune responses to mucosal pathogens. Positioning of MNPs at the epithelial interface facilitates their access to luminally-derived antigens and regulates MNP function through soluble mediators or surface receptor interactions. Therefore, accurately quantifying the distribution of MNPs within mucosal tissues as well as their spatial relationship with other cells is important to infer functional cellular interactions in health and disease. In this study, we developed and validated a MATLAB-based tissue cytometry platform, termed "MNP mapping application" (MNPmApp), that performs high throughput analyses of MNP density and distribution in the gastrointestinal mucosa based on digital multicolor fluorescence microscopy images and that integrates a Monte Carlo modeling feature to assess randomness of MNP distribution. MNPmApp identified MNPs in tissue sections of the human gastric mucosa with 98 ± 2% specificity and 76 ± 15% sensitivity for HLA-DR+ MNPs and 98 ± 1% specificity and 85 ± 12% sensitivity for CD11c+ MNPs. Monte Carlo modeling revealed that mean MNP-MNP distances for both HLA-DR+ and CD11c+ MNPs were significantly lower than anticipated based on random cell placement, whereas MNP-epithelial distances were similar to randomly placed cells. Surprisingly, H. pylori infection had no significant impact on the number of HLA-DR and CD11c MNPs or their distribution within the gastric lamina propria. However, our study demonstrated that MNPmApp is a reliable and user-friendly tool for unbiased quantitation of MNPs and their distribution at mucosal sites.
Insights
A new MATLAB tool, MNP mapping application (MNPmApp), quantifies mononuclear phagocyte (MNP) distribution in mucosal tissues. It reveals MNPs are non-randomly distributed, unaffected by H. pylori infection.
Area of Science:
- Immunology
- Cell Biology
- Computational Biology
Background:
- Mononuclear phagocytes (MNPs), including dendritic cells and macrophages, are crucial for mucosal immunity.
- Their strategic positioning at the epithelial interface influences immune responses to pathogens.
- Understanding MNP distribution and spatial relationships is key to inferring cellular interactions in health and disease.
Purpose of the Study:
- To develop and validate a MATLAB-based platform, MNP mapping application (MNPmApp), for high-throughput analysis of MNP density and distribution in gastrointestinal mucosa.
- To assess the spatial distribution patterns of MNPs using Monte Carlo modeling.
- To investigate the impact of H. pylori infection on MNP distribution in the gastric mucosa.
Main Methods:
- Development of a MATLAB-based tissue cytometry platform (MNPmApp) for analyzing digital multicolor fluorescence microscopy images.
- High-throughput quantification of MNP density and distribution in human gastric mucosa.
- Integration of Monte Carlo modeling to assess the randomness of MNP distribution and spatial relationships.
Main Results:
- MNPmApp demonstrated high specificity (98%) and sensitivity (76-85%) for identifying HLA-DR+ and CD11c+ MNPs.
- Monte Carlo modeling indicated significantly non-random MNP-MNP spacing, while MNP-epithelial distances were random.
- H. pylori infection did not significantly alter MNP numbers or distribution in the gastric lamina propria.
Conclusions:
- MNPmApp is a reliable and user-friendly tool for unbiased quantitation of MNPs and their distribution in mucosal tissues.
- MNP distribution within the gastric mucosa is non-random, suggesting specific cellular interactions.
- H. pylori infection does not appear to disrupt the fundamental distribution of key MNPs in the gastric lamina propria.
