title: Relationship of phospholipase C- and phospholipase D-mediated phospholipid remodeling pathways to respiratory burst activation in human neutrophils stimulated by Candida albicans hyphae.
abstract: Neutrophil (PMN) oxidant release, a key component of defenses against disseminated candidiasis, was preceded by oxidant generation after stimulation by Candida albicans hyphae. Opsonized or unopsonized hyphae triggered phospholipase D (PLD) activation within 5 or 30 s, respectively, forming 1-O-alkyl-phosphatidic acid (alkyl-PA) or 1-O-alkyl-phosphatidyl-ethanol in the presence of ethanol. Ethanol, which competitively lowers phosphatidic acid (PA) production, caused dose-dependent inhibition of superoxide (O2-) generation after hyphal stimulation but altered neither baseline-unstimulated O2- production nor responses to phorbol myristate acetate. PA rises evoked by unopsonized hyphae began 2 min before significant O2- release, also preceding both phospholipase C activation and cytosolic Ca2+ rises. Diacylglycerol (DAG) rose in two distinct phases after stimulation by opsonized or unopsonized hyphae, peaking briefly after 60 or 120 s, respectively, followed by prolonged secondary rises. Initial DAG rises preceded inositol triphosphate elevations evoked by unopsonized hyphae. Though PA rose before DAG, no dephosphorylation of PA to form 1-O-alkyl-DAG was noted. Propranalol, which increases PA accumulation by inhibiting PA phosphohydrolase, lowered PMN O2- responses to hyphae. Early DAG rises temporally overlapped respiratory burst initiation but PMN responses to hyphae were unchanged by a DAG kinase inhibitor, R59022, which blocks phosphorylation of DAG to PA and enhances DAG accumulation. Thus, neither PA nor DAG accumulation individually accounted for triggering PMN O2- responses to hyphae. PLD activation and PA production may facilitate PMN fungicidal responses to hyphae but play an indirect role in initiating the respiratory burst.
authors.affiliation: Section of Infectious Diseases, Department of Medicine and Evans Department of Clinical Research, Boston University Medical Center Hospital, Massachusetts.
authors.forename: T
authors.initials: T
authors.lastname: Meshulam
authors.forename: M M
authors.initials: MM
authors.lastname: Billah
authors.forename: S
authors.initials: S
authors.lastname: Eckel
authors.forename: K K
authors.initials: KK
authors.lastname: Griendling
authors.forename: R D
authors.initials: RD
authors.lastname: Diamond
mesh_terms: D002176:Candida albicans / Q000502:physiology*; D019852:Diacylglycerol Kinase; D004075:Diglycerides / Q000378:metabolism*; D000431:Ethanol / Q000494:pharmacology; D006801:Humans; D018375:Neutrophil Activation; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism*; D010712:Phosphatidic Acids / Q000378:metabolism*; D010739:Phospholipase D / Q000502:physiology*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000502:physiology; D011433:Propranolol / Q000494:pharmacology; D016897:Respiratory Burst / Q000187:drug effects; D010738:Type C Phospholipases / Q000502:physiology*
chemical_list: D004075:Diglycerides; D010712:Phosphatidic Acids; D000431:Ethanol; D011433:Propranolol; D017853:Phosphotransferases (Alcohol Group Acceptor); D019852:Diacylglycerol Kinase; D010738:Type C Phospholipases; D010739:Phospholipase D