Human macrophage adhesion on fibronectin: the role of substratum and intracellular signalling kinases.
Fibronectin and Arg-Gly-Asp (RGD)- and/or Pro-His-Ser-Arg-Asn (PHSRN)-containing oligopeptides were immobilized onto physicochemically distinct substrata: polyethyleneglycol-based networks or tissue culture polystyrene (TCPS). The role of selected signalling kinases in the adhesion of human primary blood-derived macrophages on these modified substrata was investigated. We demonstrated that the protein tyrosine kinase (PTK) or protein serine/threonine kinase (PSK) dependency and the PTK-PSK cross-talk compensation for macrophage adhesion varied dynamically with the substratum modification and the culture time. The inhibition of MAPK kinase (MAPKK) decreased macrophage adhesion on TCPS, whereas the inhibition of phosphoinositide-3 kinase (PI3 kinase) decreased macrophage adhesion on networks at 24 h. The PI3 kinase-protein kinase C (PKC)-MAPK cascade was involved in macrophage adhesion on fibronectin-preadsorbed TCPS or networks but not on fibronectin-grafted networks. This fibronectin-mediated adhesion signalling involved both RGD and PHSRN sequences in a form of G(3)RGDG(6)PHSRNG on TCPS but not on networks. Furthermore, G(3)RGDG(6)PHSRNG grafted onto networks evoked unique signalling in macrophage adhesion from that preadsorbed onto networks. Thus, macrophage adhesion and the role of selected signalling kinases were modulated by the substratum and the ligand conjugation method.
Liu
Yiping
Y
School of Pharmacy, University of Wisconsin-Madison, Madison, WI 53705, USA.
Kao
Weiyuan John
WJ
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000494:pharmacology; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005353:Fibronectins / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D008264:Macrophages / Q000502:physiology*; D009842:Oligopeptides / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D011137:Polystyrenes / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D011494:Protein Kinases / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000502:physiology; D015398:Signal Transduction
D004791:Enzyme Inhibitors; D005353:Fibronectins; D009842:Oligopeptides; D011137:Polystyrenes; D011092:Polyethylene Glycols; C047981:arginyl-glycyl-aspartic acid; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011494:Protein Kinases; D011505:Protein-Tyrosine Kinases; D011493:Protein Kinase C