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Isolation and purification of CD14-negative mucosal macrophages from normal human small intestine
P D Smith1, E N Janoff, M Mosteller-Barnum
1Department of Medicine, University of Alabama School of Medicine, Birmingham 35294, USA.
Abstract:
Mucosal macrophages play a fundamental role in the regulation of immunological events and inflammation in the small intestine. Because no information is available on normal small intestinal macrophages, we developed a technique for the isolation and purification of jejunal lamina propria macrophages in order to study their phenotype and activity. From sections of normal human jejunum, lamina propria mononuclear cells were isolated by neutral protease digestion and then subjected to counterflow centrifugal elutriation to purify the macrophages. The cells isolated by this procedure contained < 1% CD3+ lymphocytes and displayed the size distribution, morphological features, ultrastructure and phagocytic activity of mononuclear phagocytes. In contrast to blood monocytes, however, mucosal macrophages from the jejunum did not exhibit adherence properties or express CD14, a receptor for the lipopolysaccharide-binding protein. The purification of large numbers of lamina propria macrophages by this procedure offers the opportunity to define the role of this cell in the physiological inflammation characteristic of normal intestinal mucosa and the pathological inflammation associated with small intestinal diseases.
Insights
Researchers isolated jejunal lamina propria macrophages to study their function in small intestine immunity. These mucosal macrophages differ from blood monocytes, lacking CD14 expression and adherence, offering new insights into intestinal inflammation.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- Mucosal macrophages are crucial for regulating small intestinal immunity and inflammation.
- Limited information exists on the characteristics of normal small intestinal macrophages.
Purpose of the Study:
- To develop a method for isolating and purifying jejunal lamina propria macrophages.
- To investigate the phenotype and activity of these purified mucosal macrophages.
Main Methods:
- Isolation of lamina propria mononuclear cells from human jejunum via neutral protease digestion.
- Purification of macrophages using counterflow centrifugal elutriation.
- Characterization of isolated cells for purity, morphology, and phagocytic activity.
Main Results:
- A technique was established to purify jejunal lamina propria macrophages with high yield.
- Isolated cells exhibited characteristics of mononuclear phagocytes but lacked CD14 expression.
- Mucosal macrophages demonstrated different adherence properties compared to blood monocytes.
Conclusions:
- The developed method enables the study of normal small intestinal macrophages.
- Jejunal macrophages possess distinct properties compared to peripheral blood monocytes.
- This research provides a foundation for understanding intestinal inflammation in health and disease.