Oxidative metabolism of polymorphonuclear leukocytes: modulation by adhesive stimuli.
Different agents such as phorbol myristate acetate (PMA), N-formyl-methionyl-leucylphenylalanine (fMet-Leu-Phe), or opsonized zymosan induced an oxidative burst in rat peritoneal polymorphonuclear leukocytes (PMNs) elicited by casein. Plastic adhesion of PMNs down-regulated superoxide (O2) release stimulated by PMA or fMet-Leu-Phe but had no effect on zymosan-induced O2 generation, indicating that the O2 forming enzyme, the NADPH oxidase, was not affected by modulation and that a common step of the transductional events induced by PMA or fMet-Leu-Phe might be involved in this regulation. We demonstrated that a differential translocation of protein kinase C (PKC) was not responsible for that modulation. PMA-induced secretion of granule content (vitamin B12-binding protein) was not susceptible to modulation, suggesting that the transductional pathways leading to O2 generation and granule secretion are partly separated. The adhesion of PMNs to different substrates (glass, plastic, albumin-, laminin-, fibronectin-, poly-lysine-, or concanavalin A-coated plastic) down-regulated to different extent superoxide release. Whether the nature of the biochemical signal induced by the diverse adhesive stimuli or a physical parameter such as binding strength was involved in this differential behavior remains to be elucidated. Since adhesiveness was dependent on the state of the cytoskeleton and O2 inducers were reported to stimulate actin polymerization, we studied the F-actin content and distribution of PMNs by using the specific fluorescent probe NBD-phallacidin and an original methodology allowing a quantitative analysis of fluorescence on both adherent and suspended cells. PMA induced a polarization of F-actin on suspended PMNs but had no effect on the intracellular distribution of F-actin in adherent PMNs. Thus, we suggest that the adhesion of PMNs induced an immobilization of F-actin, possibly correlated to the down-regulation of one of the transductional pathways involved in the NADPH oxidase activation.
Laurent
F
F
Laboratoire d'Immunologie, Hôpital de Sainte-Marguerite, Marseille, France.
Benoliel
A M
AM
Capo
C
C
Bongrand
P
P
D000199:Actins / Q000378:metabolism; D000818:Animals; D002448:Cell Adhesion; D002451:Cell Compartmentation; D015550:Cell Degranulation; D066298:In Vitro Techniques; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate; D014155:Transcobalamins / Q000378:metabolism
D000199:Actins; D014155:Transcobalamins; D013481:Superoxides; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate