Enhanced Recruitment of CX3CR1+ T Cells by Mucosal Endothelial Cell–Derived Fractalkine in Inflammatory Bowel Disease

Background & Aims: Fractalkine (FKN/CX3CL1) is a unique chemokine combining adhesive and chemotactic properties. We investigated FKN production by the mucosal microvasculature in inflammatory bowel disease (IBD), its capacity for leukocyte recruitment into the gut, and the number of CX3CR1+ cells in the circulation and mucosa of IBD patients. Methods: The expression of FKN by human intestinal microvascular endothelial cells (HIMECs) and CX3CR1 by circulating cells was evaluated by flow cytometry, and mucosal CX3CR1+ cells were enumerated by immunohistochemistry. The capacity of FKN to mediate leukocyte binding to HIMECs was assessed by immunoblockade, and to induce HIMEC transmigration by a Transwell system. Results: The spontaneously low HIMEC FKN expression was enhanced markedly by tumor necrosis factor-α plus interferon-γ stimulation, or direct leukocyte contact. This effect was significantly stronger in IBD than control HIMECs. Up-regulation of HIMEC FKN expression was dependent on p38 and extracellular signal-regulated kinase phosphorylation, as was abrogated by selective mitogen-activated protein kinase inhibitors. Circulating T cells contained significantly higher numbers of CX3CR1+ cells in active IBD than inactive IBD or healthy subjects, and IBD mucosa contained significantly more CX3CR1+ cells than control mucosa. Antibody-blocking experiments showed that FKN was a major contributor to T- and monocytic-cell adhesion to HIMECs. Finally, FKN enhanced the expression of active β1 integrin on leukocytes and mediated leukocyte HIMEC transmigration. Conclusions: In view of the capacity of FKN to mediate leukocyte adhesion, chemoattraction, and transmigration, its increased production by mucosal microvascular cells and increased numbers of circulating and mucosal CX3CR1+ cells in IBD point to a significant role of FKN in disease pathogenesis.

Abbreviations used in this paper: Ab, antibody, ERK, extracellular signal-regulated kinase, FKN, fractalkine, HIMEC, human intestinal microvascular endothelial cell, ICAM-1, intercellular adhesion molecule 1, IFN, interferon, IL, interleukin, MAP, mitogen-activated protein, MCP-1, monocyte chemoattractant protein 1, PBMC, peripheral blood mononuclear cell, PE, phycoerythrin, TNF, tumor necrosis factor, VCAM-1, vascular cell adhesion molecule 1