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Updated: Aug 8, 2026
Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes
Published on: May 8, 2016
Flow cytometric analysis of canine colonic mucosal lymphocytes from endoscopically obtained biopsy specimens
I M Sonea1, K Harkins, M J Wannemuehler
1Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames 50011, USA.
Insights
Endoscopic colonic biopsy specimens provide a viable source of mucosal lymphocytes (ML) for flow cytometry. This minimally invasive method allows for the study of ML in client-owned animals.
Area of Science:
- Veterinary immunology
- Gastrointestinal immunology
- Canine research
Background:
- Flow cytometry is crucial for analyzing lymphocyte populations.
- Obtaining mucosal lymphocytes (ML) from canine colonic tissue can be challenging.
- Endoscopic biopsies offer a less invasive approach compared to surgical resection.
Purpose of the Study:
- To validate the use of canine colonic biopsy specimens obtained via endoscopy.
- To assess the yield and representativeness of mucosal lymphocytes (ML) for flow cytometric analysis.
- To compare ML subsets from endoscopic biopsies with those from excised colon.
Main Methods:
- Endoscopic colonic biopsy specimens were collected from adult dogs.
- Mucosal lymphocytes were isolated and processed for flow cytometry.
- Lymphocyte subsets (T cells, B cells) were identified using monoclonal antibodies.
Main Results:
- Endoscopic biopsies yielded a significant number of viable mucosal lymphocytes (ML).
- ML subsets from endoscopic biopsies were comparable to those from excised colon.
- The majority of ML obtained were intraepithelial lymphocytes.
Conclusions:
- Endoscopic colonic biopsy is a valid method for obtaining mucosal lymphocytes (ML).
- Flow cytometry using endoscopic biopsy samples is a minimally invasive technique for canine immunology studies.
- This technique facilitates research on ML in client-owned animals.
Objective:
To validate use of canine colonic biopsy specimens obtained via endoscopy as a source of mucosal lymphocytes (ML) for flow cytometric analysis.
Sample Population:
Mucosal biopsy specimens from 10 adult dogs.
Procedure:
Mucosal lymphocyte subsets obtained from excised colon were compared with ML subsets obtained from biopsy specimens obtained by use of an endoscopic forceps (6 dogs). Endoscopic colonic biopsy specimens from 4 other dogs were used to define whether obtained ML were predominantly of intraepithelial or lamina propria origin. Mucosal lymphocytes were isolated and labeled, using commercially available monoclonal antibodies directed against canine cell surface antigens. Lymphocyte subsets (cytotoxic or helper T cells; B cells) were determined by use of flow cytometric analysis.
Results:
A large number of viable ML was obtained after dissociation of the colonic epithelium from excised colon (45.5 + 21.5 X 10(6)) and endoscopic (7.2+/-3.4 X 10(6)) biopsy specimens. Lymphocyte subsets obtained with both methods were identical for each dog and consisted predominantly of intraepithelial lymphocytes, with some lymphocytes from the lamina propria. Collagenase digestion of excised colon also yielded a large number of viable lymphocytes from the lamina propria (56.7+/-20.4 X 10(6)), but collagenase digestion of endoscopic biopsy specimens was less rewarding.
Conclusion And Clinical Relevance:
A representative sample of viable intraepithelial ML is obtainable from endoscopic biopsy specimens. Flow cytometric analysis, a minimally invasive technique, can be used to study ML of client-owned animals.
