Allergen-IgE complexes trigger CD23-dependent CCL20 release from human intestinal epithelial cells.
OBJECTIVE
In food allergic individuals, exposure to food allergens by the oral route can trigger immediate (within minutes) local hypersensitivity reactions in the intestine followed by a late-phase inflammatory response. Previous work has shown that CD23 is constitutively expressed by human intestinal epithelial cells and mediates the uptake of allergen-IgE complexes. We hypothesized that allergen-IgE complexes could also signal via CD23 to trigger an inflammatory cascade in the local environment.


METHODS
Caco-2 monolayers were stimulated with human IgE-antigen (Ag) complexes. IL-8 and CCL20 mRNA and protein were determined by RT-PCR and ELISA, respectively. Signaling pathways were assessed by immunoblotting. Endogenous CD23 expression was knocked down by stable transfection with CD23 shRNA retroviral plasmid. Migration assays were performed using human monocyte-derived dendritic cells.


RESULTS
Stimulation of Caco-2 cells with IgE-Ag complexes triggered upregulation of IL-8 and CCL20 at the mRNA and protein level. Allergen complexes induced phosphorylation of ERK and JNK, but not p38 MAP kinase or NK-kappaB, and resulted in AP-1 activation. Cross-linking of CD23 replicated the findings with IgE-Ag complexes, and silencing of CD23 expression abrogated the response to allergen-IgE complexes. Supernatant from IgE-Ag-stimulated epithelial cells induced migration of dendritic cells in a CCL20-dependent manner. Finally, immunostaining of duodenal biopsies demonstrated that CCL20 was constitutively expressed by epithelial cells in vivo.


CONCLUSIONS
Signaling via epithelial CD23 may participate in the late-phase inflammatory response by the release of chemokines capable of recruiting antigen presenting cells and effector cells of allergic inflammation.
Immunology Institute, Mount Sinai School of Medicine, New York, New York, USA.
Hongxing
H
Li
Mirna
M
Chehade
Weicheng
W
Liu
Huabao
H
Xiong
Lloyd
L
Mayer
M Cecilia
MC
Berin
D000485:Allergens / Q000276:immunology*; D018938:Caco-2 Cells; D054418:Chemokine CCL20 / Q000276:immunology*; D004847:Epithelial Cells / Q000276:immunology; D005512:Food Hypersensitivity / Q000276:immunology; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology*; D007413:Intestinal Mucosa / Q000276:immunology*; D019402:Macrophage Inflammatory Proteins / Q000276:immunology*; D017455:Receptors, IgE / Q000276:immunology*; D015398:Signal Transduction
D000485:Allergens; C108355:CCL20 protein, human; D054418:Chemokine CCL20; D019402:Macrophage Inflammatory Proteins; D017455:Receptors, IgE; D007073:Immunoglobulin E