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Updated: Aug 8, 2026
Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine
Published on: May 21, 2012
Immunolocalization of CD1d in human intestinal epithelial cells and identification of a
K Somnay-Wadgaonkar1, A Nusrat, H S Kim
1Department of Medicine, Brigham & Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
Insights
Researchers identified CD1d glycoproteins on human intestinal epithelial cells, localizing them to apical and lateral surfaces. This strategic positioning suggests a role in sampling luminal antigens for immune presentation.
Area of Science:
- Immunology
- Gastroenterology
- Cell Biology
Background:
- CD1d molecules present lipid antigens to T cells.
- Understanding CD1d's role in the intestine is crucial for mucosal immunity.
Purpose of the Study:
- To define the cellular localization and biochemical structure of CD1d in human intestinal epithelial cells (IEC).
Main Methods:
- Immunoprecipitation and Western blotting using CD1d-specific antibodies.
- N-glycanase digestion to analyze protein glycosylation.
- Confocal laser microscopy for immunolocalization.
- Selective cell surface biotinylation.
Main Results:
- Identified a 37 kDa non-glycosylated and a 48-50 kDa beta2-microglobulin-associated CD1d glycoprotein in IEC.
- Demonstrated CD1d localization to both apical and lateral surfaces of intestinal epithelial cells.
- Revealed a significant intracellular apical pool of CD1d.
- Confirmed glycosylated CD1d on both apical and basolateral surfaces, with the non-glycosylated form on the apical surface.
Conclusions:
- CD1d is strategically localized on intestinal epithelial cells for luminal antigen sampling.
- The identified CD1d forms and locations support its role in presenting antigens to intraepithelial lymphocytes.
Abstract:
In order to better understand the role of intestinal CD1d, we sought to define the cellular localization and further characterize the biochemical structure of CD1d in human intestinal epithelial cells (IEC). Using a CD1d-specific rabbit anti-gst-CD1d antibody, immunoprecipitation of radiolabeled cell surface proteins detected a previously identified 37 kDa protein as well as a 48-50 kDa protein which were confirmed by Western blotting with a CD1d-specific mAb, D5. Immunoprecipitation of protein lysates with the CD1d-specific mAb, D5 and 51.1.3, and the beta2-microglobulin (beta2m)-specific mAb, BBM.1, followed by N-glycanase digestion and Western blotting with the D5 mAb showed that the 48-50 kDa protein was a beta2m-associated, CD1d glycoprotein. CD1d was immunolocalized to the apical and lateral regions of native small and large intestinal IEC as defined by confocal laser microscopy using the D5 mAb and the rabbit anti-gst-CD1d antibody. In addition, a large apical intracellular pool of CD1d was identified. Identical observations were made with polarized T84 cells. Selective biotin labeling of apical and basolateral cell surfaces followed by immunoprecipitation with the D5 mAb, N-glycanase digestion and avidin blotting confirmed the presence of glycosylated CD1d on both cell surfaces and immunolocalization of the 37 kDa non-glycosylated form of CD1d to the apical cell surface. These studies show that CD1d is located in an ideal position for luminal antigen sampling and presentation to subjacent intraepithelial lymphocytes.
