title: Migration of a human keratinocyte cell line (HACAT) to interstitial collagen type I is mediated by the alpha 2 beta 1-integrin receptor.
abstract: The migratory response of the human keratinocyte cell line HaCaT to collagen type I and the molecular mechanism underlying collagen-mediated migration have been analyzed. The migratory response of HaCaT cells to collagen type I consisted of a dose-dependent migration to insoluble step gradients of substratum-bound collagen (haptotaxis) and to gradients of soluble collagen (chemotaxis). Checkerboard analysis demonstrated a minor chemokinetic component. Denatured collagen type I was less chemoattractive than the native triple-helical form. Pre-treatment of cells with 25-250 micrograms/ml of synthetic peptides containing the fibronectin cell-recognition sequence RGD (Arg-Gly-Asp) resulted in a concentration-dependent inhibition of fibronectin-mediated chemotaxis, whereas chemotaxis to collagen was not affected. We then investigated the role of VLA/collagen-receptors for collagen type I-induced chemotaxis. Monoclonal antibody (MoAb) 5E8, which selectively blocks function of the alpha 2 subunit of the VLA-2/collagen receptor, dose-dependently inhibited the chemotactic response of HaCaT cells to collagen. This effect was specific for collagen-mediated chemotaxis because the chemotactic response to fibronectin remained unaffected. In contrast, a function blocking MoAb directed to the alpha 3 subunit of the coexpressed VLA-3 receptor, which is also capable of binding collagen, had no effect. However, function blocking MoAb directed to the beta 1-chain of integrins completely inhibited chemotaxis to collagen type I. Based on our results, we propose that the chemotactic migration of the human keratinocyte cell line (HaCaT) to collagen type I is specifically mediated by the RGD independent VLA-2/collagen receptor (alpha 2 beta 1) of the integrin family.
authors.affiliation: Department of Dermatology, Universities of Düsseldorf, F.R.G.
authors.forename: K
authors.initials: K
authors.lastname: Scharffetter-Kochanek
authors.forename: C E
authors.initials: CE
authors.lastname: Klein
authors.forename: G
authors.initials: G
authors.lastname: Heinen
authors.forename: C
authors.initials: C
authors.lastname: Mauch
authors.forename: T
authors.initials: T
authors.lastname: Schaefer
authors.forename: B C
authors.initials: BC
authors.lastname: Adelmann-Grill
authors.forename: G
authors.initials: G
authors.lastname: Goerz
authors.forename: N E
authors.initials: NE
authors.lastname: Fusenig
authors.forename: T M
authors.initials: TM
authors.lastname: Krieg
authors.forename: G
authors.initials: G
authors.lastname: Plewig
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology; D002460:Cell Line; D002465:Cell Movement; D002633:Chemotaxis; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D015603:Keratinocytes / Q000502:physiology*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000494:pharmacology; D016029:Receptors, Very Late Antigen / Q000276:immunology / Q000502:physiology*
chemical_list: D000911:Antibodies, Monoclonal; D009842:Oligopeptides; D016029:Receptors, Very Late Antigen; C047981:arginyl-glycyl-aspartic acid; D003094:Collagen