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Detection of small intestinal IgA+TNFα+iNOS+ CELLS by Immunofluorescence Microscopy (IF)

External protocol Created on 30 Apr 2014

Authors

Jennifer Gommerman and Jorg Fritz

Summary

The largest mucosal surface in the body is in the gastrointestinal (GI) tract, a location that is heavily colonized by normally harmless microbes. A key mechanism required for maintaining a homeostatic balance between this microbial burden and the lymphocytes that densely populate the GI tract is the production and trans-epithelial transport of poly-reactive IgA1. Within the mucosal tissues, B cells respond to cytokines, sometimes in the absence of T cell help, undergo class switch recombination (CSR) of their Immunoglobulin (Ig) receptors to IgA, and differentiate to become plasma cells (PC)2. However, IgA-secreting PC likely have additional attributes that are needed for coping with the tremendous bacterial load in the GI tract. We report that IgA+ PC also can produce the anti-microbial mediators TNFα and iNOS, which appear to arise in the unique environment of the gut, and may be critical to mount effective responses to microbial assault. Here we described a detailed method to characterize and quantify IgA+iNOS+TNFα+ cells that we called TNFα-iNOS-producing (Tip)-PC in the lamina propria of mice by immunohistochemistry (IHC).

Further details

The protocol was published on Protocol Exchange on 16 December 2011. To see the entire protocol, click on the source link.

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