Mechanisms of beta 1 integrin-dependent adherence of granulocytic HL60 to fibronectin.
We investigated the mechanism of beta 1 integrin-mediated adherence of stimulated granulocytic HL60 cells to fibronectin using a monoclonal antibody (15/7) that recognizes beta 1 integrins only when the receptors are active for ligand binding. Phorbol myristate acetate (PMA) stimulated expression of the 15/7 epitope on granulocytic HL60 by nearly fivefold but had an insignificant effect on the expression of the epitope on undifferentiated HL60 cells. These results paralleled the effect of PMA on HL60 and granulocytic HL60 adhesion to fibronectin, indicating that activation of beta 1 integrins is important for beta 1-mediated adherence of granulocytic HL60 cells to fibronectin. Agonists that stimulate alpha 5 beta 1-dependent human polymorphonuclear leukocyte (PMN) adhesion to fibronectin (C5a and PMA) also upregulated the 15/7 epitope on purified human PMNs. Although PMA rapidly induces increased levels of filamentous actin (F-actin) in granulocytic HL60 cells and a decrease in F-actin levels in undifferentiated HL60 cells, depolymerization of the actin cytoskeleton with cytochalasin B did not affect increased expression of the 15/7 epitope on granulocytic HL60 cells. Cytochalasin B did, however, inhibit granulocytic HL60 adherence to fibronectin by 50%, demonstrating that actin polymerization is important for optimal beta 1-dependent granulocytic adherence.
Bohnsack
J F
JF
Department of Pediatrics, University of Utah Health Sciences Center, Salt Lake City 84132, USA.
Chang
J
J
Zhou
X
X
Yednock
T A
TA
D000199:Actins / Q000378:metabolism; D000328:Adult; D002448:Cell Adhesion / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D015936:Complement C5a / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D000939:Epitopes; D005353:Fibronectins / Q000502:physiology*; D006098:Granulocytes / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D019012:Integrin beta1; D016023:Integrins / Q000502:physiology*; D007951:Leukemia, Myeloid / Q000473:pathology*; D013268:Stimulation, Chemical; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D015854:Up-Regulation / Q000187:drug effects
D000199:Actins; D000939:Epitopes; D005353:Fibronectins; D019012:Integrin beta1; D016023:Integrins; D003571:Cytochalasin B; D015936:Complement C5a; D013755:Tetradecanoylphorbol Acetate