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Isolation and functional characteristics of adherent phagocytic cells from mouse Peyer's patches
Abstract:
Attempts were made to isolate adherent phagocytic cells (macrophages) from mouse Peyer's patch cell suspensions. Cell suspensions prepared by teasing apart the Peyer's patches contained no adherent phagocytic cells. However, if Peyer's patch fragments were treated with collagenase to disrupt the tissue matrix, cells prepared in this way contained a subpopulation of adherent phagocytic cells. These cells comprised only 0.1-0.2% of the total nucleated cell population of the Peyer's patch. Similar cells could also be isolated from the Peyer's patches of germ-free mice, but as judged by their ability to ingest opsonized erythrocytes, these cells were less activated than cells from the Peyer's patches of normal mice. Adherent cells from the Peyer's patches of normal mice could present antigen (ovalbumin) to T cells, and Peyer's patches cell suspensions containing adherent cells could be stimulated in vitro to produce an anti-sheep red blood cell plaque-forming cell response in the absence of 2-mercaptoethanol. These studies show that although the frequency of phagocytic adherent cells is extremely low in Peyer's patches, these cells have functions consistent with that of adherent cells in other lymphoid tissues.
Insights
Macrophages are rare in mouse Peyer's patches but can be isolated using collagenase. These phagocytic cells, though infrequent, demonstrate antigen presentation and immune response functions, similar to those in other lymphoid tissues.
Area of Science:
- Immunology
- Gastrointestinal Biology
Background:
- Peyer's patches are crucial immune sites in the gut.
- Adherent phagocytic cells, such as macrophages, play key roles in immune responses.
Purpose of the Study:
- To investigate the presence and function of adherent phagocytic cells in mouse Peyer's patches.
- To characterize these cells and their immune capabilities.
Main Methods:
- Isolation of cells from Peyer's patch fragments treated with collagenase.
- Assessment of phagocytic activity using opsonized erythrocytes.
- Evaluation of antigen presentation to T cells.
- In vitro stimulation for plaque-forming cell response.
Main Results:
- Adherent phagocytic cells were successfully isolated after collagenase treatment of Peyer's patch fragments.
- These cells represented a very small fraction (0.1-0.2%) of the total nucleated cells.
- Cells from germ-free mice showed lower activation levels compared to normal mice.
- Isolated cells demonstrated antigen presentation and supported in vitro immune responses.
Conclusions:
- Despite their low frequency, Peyer's patch macrophages possess functional characteristics similar to macrophages in other lymphoid organs.
- Collagenase digestion is essential for isolating these adherent cells from the Peyer's patch tissue matrix.