Regulation of interleukin-8 receptor expression in human polymorphonuclear neutrophils.
Interleukin-8, a neutrophil chemotactic agent, is known to have an active role in the induction of inflammatory response in a number of diseases. Although the activity of IL-8 is known to be through a receptor (IL-8R) on the surface of neutrophils, no information is available regarding the regulation of the IL-8R expression. The present study demonstrates that serum activated LPS at a concentration of 10 ng/ml induces expression of functionally active IL-8R by 120% within 30 min through de novo protein synthesis. The upregulated receptors could be detected by anti-IL-8R antibody and could also be demonstrated by autoradiography with crosslinking 125I IL-8. The serum-activated LPS-stimulated neutrophils migrated faster and showed higher Ca2+ flux over the unstimulated cells. The LPS-induced receptors were downregulated rapidly, about 85% of the receptor activity being lost within 90 min of incubation at 37 degrees C. The downregulation could be partially prevented by treatment with a cocktail of protease inhibitors, suggesting the possible involvement of protease(s) in this process. Both EDTA (100 microM) and bestatin (40 microM) afforded almost complete protection of the receptor from proteolytic cleavage indicating that the enzyme involved is a metalloprotease, possibly an aminopeptidase. The study shows that stimulation of PMNs with LPS leads to induction of IL-8R expression enhancing the IL-8-mediated biological responses and also provides evidence for post-stimulatory restoration of receptor level on the neutrophil surface by proteolytic cleavage of the amino-terminal end of the receptor.
Manna
S K
SK
Division of Immunobiology, Indian Institute of Chemical Biology, Calcutta.
Bhattacharya
C
C
Gupta
S K
SK
Samanta
A K
AK
D015703:Antigens, CD / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002634:Chemotaxis, Leukocyte; D015536:Down-Regulation; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007249:Inflammation / Q000209:etiology; D016209:Interleukin-8 / Q000378:metabolism*; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology* / Q000502:physiology*; D011480:Protease Inhibitors / Q000494:pharmacology; D018123:Receptors, Interleukin / Q000187:drug effects / Q000378:metabolism*; D023062:Receptors, Interleukin-8A; D015854:Up-Regulation
D015703:Antigens, CD; D016209:Interleukin-8; D008070:Lipopolysaccharides; D011480:Protease Inhibitors; D018123:Receptors, Interleukin; D023062:Receptors, Interleukin-8A; D010450:Endopeptidases; D002118:Calcium