Migration of eosinophils through basement membrane components in vitro: role of matrix metalloproteinase-9.
In general, inflammatory cells cross basement membranes by producing proteinases. To investigate the role of proteinases in eosinophil basement membrane migration, we studied peripheral blood eosinophils in Matrigel-coated chemotaxis chambers. Electron microscopy showed degradation of the Matrigel layer when eosinophils, added to the upper chamber, transmigrated the membrane in the presence of both platelet-activating factor (PAF) in the lower chamber and interleukin (IL)-5 in both chambers. In the absence of either or both PAF and IL-5, no changes occurred in the Matrigel layer. Matrigel transmigration of eosinophils induced by PAF and IL-5 was inhibited by 1,10-phenanthroline, batimastat, 3,4-dichloroisocoumarin, chymostatin, and a neutralizing antibody for the matrix metalloproteinase (MMP)-9, indicating that serine proteinase(s) and MMP, specifically MMP-9, were involved in the transmigration of eosinophils through Matrigel. In contrast, eosinophil migration through a bare membrane was not affected by batimastat. Using gelatin zymography and immunoblotting, MMP-9 was detected in the migration upper chamber supernatant of the eosinophil transmigration assay and in the conditioned medium of eosinophils. Release of MMP-9 by eosinophils was increased by IL-5, PAF, or both, but the substrate-degrading activity of MMP-9 was increased only in the presence of both IL-5 and PAF, indicating that the releasing and activating mechanisms of MMP-9 are involved in eosinophil basement membrane migration. This study implicates MMP-9 in basement membrane migration of eosinophils and suggests its involvement in inflammatory diseases where tissue eosinophilia plays a role.
Okada
S
S
Department of Immunology, Mayo Clinic and Mayo Foundation, Rochester, Minnesota 55905, USA.
Kita
H
H
George
T J
TJ
Gleich
G J
GJ
Leiferman
K M
KM
D001485:Basement Membrane / Q000166:cytology* / Q000378:metabolism; D002465:Cell Movement; D002478:Cells, Cultured; D017364:Collagenases / Q000378:metabolism*; D004804:Eosinophils / Q000166:cytology*; D006801:Humans; D015848:Interleukin-5 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9; D008855:Microscopy, Electron, Scanning; D010972:Platelet Activating Factor / Q000378:metabolism
D015848:Interleukin-5; D010972:Platelet Activating Factor; D017364:Collagenases; D020780:Matrix Metalloproteinase 9