Monocyte adherence to the subendothelial basement membrane increases interleukin-8 gene expression and antigen release.
The emigration of peripheral blood monocytes into the interstitium allows for contact with a variety of surfaces which may provide signals important for monocyte function in both normal and inflammatory states. In the present study, we examined the effect of adherence to an endothelial cell-derived basement membrane and to collagen I, the major collagen of the interstitium, on monocyte release and gene expression of the potent chemotactic cytokine Interleukin-8 (IL-8). We further evaluated neutrophil chemotactic activity of the conditioned media containing antigenic IL-8 from monocytes adherent to these same surfaces. Elutriation-purified monocytes were adhered for 1 hour to plastic tissue culture wells either uncoated (PL) or coated with bovine serum albumin (BSA), collagen type I (C-I), or endothelial cell-derived basement membrane (BM). Following removal of nonadherent cells, monocytes were further incubated in a serum-free media for 18 hours in the presence or absence of lipopolysaccharide (IPS). Following 18 hrs of incubation there were significantly less monocytes remaining adherent to BM when compared to other surfaces tested. In the absence of LPS, adherent monocytes released significant amounts of IL-8 that was not surface specific. In the presence of LPS, monocytes adherent to BM released significantly more IL-8, when corrected for adherent cell number, than monocytes adherent to PL, BSA, or C-I. Conditioned media from adherent monocytes expressed IL-8 dependent neutrophil chemotactic activity that was not influenced by the surfaces tested. Northern blot analysis indicated greater induction for IL-8 mRNA by monocytes adhered to BM after 18 hrs in the presence of LPS. These results suggest that monocyte adherence to the subendothelial basement membrane provides a priming signal for the induction and secretion of the chemotactic cytokine IL-8 in response to inflammatory stimuli.
Department of Physiology and Cell Biology, Albany Medical College, New York 12208, USA.
L A
LA
Heinel
D
D
Singleton
M
M
Miller
M B
MB
Frewin
P W
PW
Gudewicz
D000941:Antigens / Q000378:metabolism; D001483:Base Sequence; D001485:Basement Membrane / Q000502:physiology*; D002448:Cell Adhesion; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D015870:Gene Expression; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics* / Q000276:immunology / Q000378:metabolism*; D015335:Molecular Probes / Q000235:genetics; D008969:Molecular Sequence Data; D009000:Monocytes / Q000502:physiology*
D000941:Antigens; D016209:Interleukin-8; D015335:Molecular Probes