Circulating growth-regulator oncogene alpha contributes to neutrophil priming and interleukin-8-directed mucosal recruitment into chronic lesions of patients with Crohn's disease.
Crohn's disease (CD) lesions are characterized by a marked neutrophilic infiltrate associated with enhanced mucosal IL-8, contrasting with low serum IL-8 levels. The aim of this study was to investigate the effects of circulating GROalpha and IL-8 on neutrophil priming and migration. The expression of surface molecules involved in rolling (CD62L, CD15) and firm adhesion (Mac-1 and LFA-1) to endothelial cells was assessed by flow cytometry, while the chemotactic response of PMN to IL-8 and to fMLP was investigated in a Boyden chamber assay. In addition, IL-8 and GROalpha levels were determined by ELISA in plasma samples and in culture supernatants of purified polymorphonuclear neutrophils (PMN) and peripheral blood mononuclear cells (PBMC) from patients with CD and healthy blood donors. This study revealed an upregulation of CD11b (Mac-1) membrane expression on circulating PMN from patients with CD, as assessed by the mean fluorescence intensity which reflects antigen density. Furthermore, an enhanced chemotactic response towards both fMLP and IL-8 of PMN from CD patients was observed. Despite often undetectable levels of circulating IL-8, all plasma samples were positive for GROalpha, with a significant increase in CD patients when compared to donors. In vitro, equivalent concentrations of GROalpha were able to increase the IL-8 driven chemotaxis of PMN. In conclusion, blood PMN from patients with CD showed an enhanced capacity to be recruited into inflammed intestinal mucosa, which could be due to an increased expression of CD11b (Mac-1) as well as an increased chemotactic response toward fMLP or IL-8. This priming effect of PMN in CD may partly occur through elevated circulating GROalpha levels.
Brandt
E
E
Unité INSERM U. 167, Institut Pasteur de Lille, BP 245, 59019 Lille Cedex, France.
Müller-Alouf
H
H
Desreumaux
P
P
Woerly
G
G
Colombel
J F
JF
Capron
M
M
D016022:Case-Control Studies; D002448:Cell Adhesion; D002465:Cell Movement; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000097:blood* / Q000502:physiology; D002634:Chemotaxis, Leukocyte / Q000276:immunology / Q000502:physiology; D003424:Crohn Disease / Q000276:immunology* / Q000473:pathology / Q000503:physiopathology; D006133:Growth Substances / Q000097:blood* / Q000502:physiology; D006801:Humans; D018928:Immunity, Mucosal; D066298:In Vitro Techniques; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8 / Q000494:pharmacology* / Q000502:physiology; D019041:L-Selectin / Q000378:metabolism; D016256:Lewis X Antigen / Q000378:metabolism; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism; D016177:Macrophage-1 Antigen / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology* / Q000502:physiology
C475829:CXCL1 protein, human; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D006133:Growth Substances; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8; D016256:Lewis X Antigen; D016169:Lymphocyte Function-Associated Antigen-1; D016177:Macrophage-1 Antigen; D019041:L-Selectin; D009240:N-Formylmethionine Leucyl-Phenylalanine