Gastroenterology

Gastroenterology

Volume 126, Issue 1, January 2004, Pages 63-80
Gastroenterology

Clinical-alimentary tract
CD40-mediated immune-nonimmune cell interactions induce mucosal fibroblast chemokines leading to T-Cell transmigration

https://doi.org/10.1053/j.gastro.2003.10.046Get rights and content

Abstract

Background & Aims: The CD40 pathway is a key mediator of inflammation and autoimmunity. We investigated cell adhesion molecule (CAM) up-regulation and chemokine production by CD40-positive human intestinal fibroblasts (HIF) and microvascular endothelial cells (HIMEC) induced by CD40 ligand (CD40L)-positive T cells and soluble CD40L and their effect on T-cell adhesion and transmigration. Methods: Expression of CD40, CD40L, and CAM was assessed by immunohistochemistry, confocal microscopy and flow cytometric analysis, and chemokine production using enzyme-linked immunosorbent assay. Calcein-labeled T cells were used to assay HIF adhesion and Transwell HIMEC transmigration. Results: Ligation of CD40-positive HIF and HIMEC by CD40L-positive T cells or soluble CD40L induced up-regulation of CAM expression as well as interleukin-8 and RANTES production. The specificity of these responses was shown by inhibition with a CD40L blocking antibody and by CD40 signaling-dependent p38 mitogen-activated protein kinase phosphorylation. On CD40 ligation, HIF increased their T-cell binding capacity and generated chemoattractants able to induce T-cell migration through HIMEC monolayers. Conclusions: Activation of the CD40/CD40L system in the gut mucosa may trigger a self-sustaining loop of immune-nonimmune cell interactions leading to an antigen-independent influx of T cells that contributes to chronic inflammation.

Section snippets

Isolation and cultures of HIF and HIMEC

Surgical specimens used for isolation of HIF and HIMEC were all of colonic origin. Mucosal strips were obtained from histologically normal tissue of patients admitted for bowel resection due to malignant and nonmalignant conditions, including colon cancer, benign polyps, and diverticulosis, or tissue involved by CD or UC. All diagnoses were confirmed by clinical, radiologic, endoscopic, and histologic criteria. Isolation of HIF and HIMEC was performed as previously reported.31, 32 Briefly, HIF

Expression of CD40 and CD40L in normal and IBD-involved colonic mucosa

Immunohistochemistry for CD40 of histologically normal colonic tissue from control subjects showed staining of some mononuclear cells in the lamina propria and weak staining of microvascular endothelial cells in the submucosa (Figure 1A). Muscularis mucosae and mesenchymal cells were negative for CD40. In actively inflamed CD tissue, there was strong and diffuse CD40 staining of mononuclear, endothelial, and mesenchymal cells in the mucosa and submucosa (Figure 1B), and the same cells were

Discussion

The results of this study show that the cognate interaction of mucosal CD40-positive nonimmune cells with CD40L-positive T cells results in the up-regulation of CAM by HIF, which increase their T-cell binding capacity and secretion of biologically active chemoattractants able to induce migration of T cells through a HIMEC monolayer. This series of events is likely to recapitulate what occurs in vivo given the enhanced expression of CD40 and CD40L by nonimmune and immune cells, respectively, in

Acknowledgements

The authors thank the Human Tissue Procurement Facility of University Hospitals of Cleveland (Cleveland, OH) for supplying tissue samples and Immunex Corp. (Seattle, WA) for providing essential reagents.

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    Supported by grants from the National Institutes of Health (NRSA DK10041 to J.D.V., DK30399 and DK50984 to C.F., and DK58867 to S.A.S).

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