Lymphotoxin induces secretion of granule proteins from adherent neutrophils: possible role of intracellular free calcium.
Lymphotoxin (LT) can activate human neutrophils. Using a hemolytic plaque assay to detect secretion of lactoferrin and myeloperoxidase (MPO) from single adherent neutrophils, we showed that LT induced secretion from both primary and secondary granules. Incubation of cells with cytochalasin B was required for MPO secretion, and it enhanced lactoferrin secretion. Pertussis toxin, which blocks a G-protein in the plasma membrane, inhibited LT-induced exocytosis of MPO, but not of lactoferrin. Incubation with LT did not induce any detectable changes of the cytoplasmic free [Ca2+] in neutrophils. On the other hand, secretion of granule proteins from adherent neutrophils in response to LT was blocked by loading neutrophils with quin-2 in order to increase the intracellular calcium buffering capacity. This was achieved at a concentration of quin-2, at which the secretion induced by the phorbol ester PMA and the chemotactic peptide FMLP was unaffected. Trifluoroperazine (TFP), a dual protein kinase C and calmodulin inhibitor, significantly inhibited the LT-mediated secretion of lactoferrin from adherent granulocytes. The PMA effect was unaltered by TFP under these conditions, suggesting that the inhibitory effect was on a calcium-calmodulin dependent step. The secretion induced by TNF and GM-CSF was also blocked by buffering changes in the intracellular [Ca2+] and inhibited to a similar extent by TFP. Our results suggest that calmodulin and minute changes in the cytoplasmic free [Ca2+] may be involved in a common signal transduction pathway engaged in activation of adherent neutrophils by several cytokines.
Richter
J
J
Department of Medicine, University Hospital, Lund, Sweden.
D002118:Calcium / Q000032:analysis / Q000502:physiology; D002448:Cell Adhesion / Q000502:physiology; D015550:Cell Degranulation / Q000187:drug effects / Q000502:physiology; D003594:Cytoplasmic Granules / Q000032:analysis / Q000378:metabolism*; D003600:Cytosol / Q000032:analysis; D006801:Humans; D007781:Lactoferrin / Q000032:analysis / Q000378:metabolism*; D007782:Lactoglobulins / Q000378:metabolism*; D008233:Lymphotoxin-alpha / Q000494:pharmacology*; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D009195:Peroxidase / Q000032:analysis / Q000378:metabolism*; D037342:Pertussis Toxin; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014268:Trifluoperazine / Q000494:pharmacology; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D007782:Lactoglobulins; D008233:Lymphotoxin-alpha; D010566:Virulence Factors, Bordetella; D014268:Trifluoperazine; D009195:Peroxidase; D037342:Pertussis Toxin; D007781:Lactoferrin; D002118:Calcium