Extracellular matrix proteins attenuate activation and degranulation of stimulated eosinophils.
Cellular adhesion plays an important role in the recruitment and activation of eosinophils. Here, we investigated whether extracellular matrix (ECM) proteins modify effector functions of activated human eosinophils. We coated 96-well plates with laminin or fibronectin and blocked nonspecific protein-binding sites with human serum albumin (HSA). When eosinophils were stimulated with platelet-activating factor (PAF) and incubated in these ECM-coated wells, the eosinophils adhered using both beta 1- and beta 2-integrins. Degranulation of eosinophils adherent to laminin- and fibronectin-coated wells was reduced about 50% compared with cells adherent to uncoated, HSA-blocked wells. Furthermore, these inhibitory effects of laminin and fibronectin were concentration-dependent and secretagogue-specific, that is, degranulation induced by C5a and IL-5 was inhibited while degranulation induced by secretory IgA and PMA was not inhibited. Plasma fibronectin, type I collagen and type IV collagen also inhibited PAF- and C5a-induced eosinophil degranulation, whereas fibrinogen did not. By microscopy, PAF-stimulated eosinophils, adhering to uncoated HSA-blocked wells, appeared elongated with many pseudopods. In contrast, eosinophils adhering to laminin-coated wells appeared oval with few pseudopods. Furthermore, when cells were incubated in laminin-coated wells, PAF-stimulated production of a second messenger, inositol phosphate, was markedly reduced. These findings suggest that ECM protein, such as laminin and fibronectin, attenuate both activation and degranulation of eosinophils and also influence their morphology after stimulation by physiologic secretagogues. Thus, ECM proteins may regulate activation of eosinophils, as they traverse between the peripheral blood and their targets.
Kita
H
H
Department of Immunology, Mayo Clinic, Rochester, MN 55905, USA.
Horie
S
S
Gleich
G J
GJ
D002448:Cell Adhesion / Q000187:drug effects; D015550:Cell Degranulation / Q000187:drug effects*; D048429:Cell Size / Q000187:drug effects; D004804:Eosinophils / Q000187:drug effects* / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000097:blood / Q000494:pharmacology*; D005353:Fibronectins / Q000097:blood / Q000494:pharmacology; D006801:Humans; D007295:Inositol Phosphates / Q000096:biosynthesis / Q000097:blood; D007797:Laminin / Q000097:blood / Q000494:pharmacology; D010972:Platelet Activating Factor / Q000494:pharmacology
D016326:Extracellular Matrix Proteins; D005353:Fibronectins; D007295:Inositol Phosphates; D007797:Laminin; D010972:Platelet Activating Factor