Intestinal heat shock protein 110 regulates expression of CD1d on intestinal epithelial cells
Sean P. Colgan, … , John R. Subjeck, Richard S. Blumberg
Sean P. Colgan, … , John R. Subjeck, Richard S. Blumberg
Published September 1, 2003
Citation Information: J Clin Invest. 2003;112(5):745-754. https://doi.org/10.1172/JCI17241.
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Article Immunology

Intestinal heat shock protein 110 regulates expression of CD1d on intestinal epithelial cells

Abstract

CD1d is expressed on the surface of professional and nonprofessional APCs, including intestinal epithelial cells (IECs), for a role in the presentation of glycolipid-based antigens to subsets of T cells. The mechanisms that regulate CD1d expression in any cell type are unknown. To investigate the possibility that expression of CD1d is influenced by exogenous factors present within the intestinal lumen, CD1d expression was analyzed in several IEC lines after culturing in the presence of lumenal contents (LC) of the normal human intestine. Exposure of the colon-derived cell lines T84, HT-29, and Caco-2 to soluble LC resulted in a marked induction of CD1d expression as determined by RT-PCR, confocal microscopy, cell surface ELISA, and Western blot analysis. Similarly, exposure of human IECs to LC isolated from mice bred in both specific pathogen–free and germfree conditions also resulted in the induction of CD1d expression, with the maximum CD1d-inducing activity observed in the small intestine. Biochemical and biophysical characterization of the human CD1d-inducing activity identified heat shock protein 110 (Hsp110) as a major functional component of the LC that contributes to CD1d surface regulation, and immunolocalization studies revealed Hsp110 expression in subsets of human IECs in vivo. These data support the presence of a novel autocrine pathway of CD1d regulation by Hsp110.

Authors

Sean P. Colgan, Richard S. Pitman, Takashi Nagaishi, Atsushi Mizoguchi, Emiko Mizoguchi, Lloyd F. Mayer, Ling Shao, R. Balfour Sartor, John R. Subjeck, Richard S. Blumberg

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Figure 3

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Soluble LC material elevates CD1d expression in the colonic epithelium-d...
Soluble LC material elevates CD1d expression in the colonic epithelium-derived cell lines HT-29 and Caco-2. (a) CD1d-specific cell surface ELISA of confluent monolayers of HT-29 cells following incubation with LC samples. As a positive control, cells were exposed to IFN-γ at a final concentration of 1,000 IU/ml for a period of 48 hours. (b) Western blot of whole-cell lysates isolated from HT-29 and Caco-2 cells following treatment with soluble LC material at a concentration of 6 μg/ml. Proteins contained within the lysates were resolved by 12% SDS-PAGE and transferred to nitrocellulose. The presence of CD1d was subsequently assessed by probing with the anti-human CD1d-specific antibody D5. CD1d was observed as dominant bands of 48-kDa (filled arrowhead) and 37-kDa (open arrowhead) forms. No bands were detected when nitrocellulose membranes were probed with normal mouse serum (data not shown). The blots were reprobed for the presence of β-actin (bottom panels).