Functional coupling of FcgammaRI to nicotinamide adenine dinucleotide phosphate (reduced form) oxidative burst and immune complex trafficking requires the activation of phospholipase D1.
Immunoglobulin G (IgG) receptors (FcgammaRs) on myeloid cells are responsible for the internalization of immune complexes. Activation of the oxidase burst is an important component of the integrated cellular response mediated by Fc receptors. Previous work has demonstrated that, in interferon-gamma-primed U937 cells, the high-affinity receptor for IgG, FcgammaRI, is coupled to a novel intracellular signaling pathway that involves the sequential activation of phospholipase D (PLD), sphingosine kinase, and calcium transients. Here, it is shown that both known PLD isozymes, PLD1 and PLD2, were present in these cells. With the use of antisense oligonucleotides to specifically reduce the expression of either isozyme, PLD1, but not PLD2, was found to be coupled to FcgammaRI activation and be required to mediate receptor activation of sphingosine kinase and calcium transients. In addition, coupling of FcgammaRI to activation of the nicotinamide adenine dinucleotide phosphate (reduced form) (NADPH) oxidase burst was inhibited by pretreating cells with 0.3% butan-1-ol, indicating an absolute requirement for PLD. Furthermore, use of antisense oligonucleotides to reduce expression of PLD1 or PLD2 demonstrated that PLD1 is required to couple FcgammaRI to the activation of NADPH oxidase and trafficking of internalized immune complexes for degradation. These studies demonstrate the critical role of PLD1 in the intracellular signaling cascades initiated by FcgammaRI and its functional role in coordinating the response to antigen-antibody complexes.
Melendez
A J
AJ
Department of Molecular and Cellular Biology, Pfizer Global Research and Development, Fresnes, France.
Bruetschy
L
L
Floto
R A
RA
Harnett
M M
MM
Allen
J M
JM
D000936:Antigen-Antibody Complex / Q000378:metabolism*; D003994:Bucladesine / Q000494:pharmacology; D000440:Butanols / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D004789:Enzyme Activation; D006801:Humans; D008264:Macrophages / Q000502:physiology; D009000:Monocytes / Q000166:cytology / Q000502:physiology; D009249:NADP / Q000378:metabolism*; D019255:NADPH Oxidases / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D010084:Oxidation-Reduction; D010739:Phospholipase D / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000378:metabolism; D017452:Receptors, IgG / Q000378:metabolism*; D016897:Respiratory Burst; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D000099157:Sphingosine Kinase
D000936:Antigen-Antibody Complex; D003994:Bucladesine; D000440:Butanols; D002118:Calcium; D009249:NADP; D019255:NADPH Oxidases; D016376:Oligonucleotides, Antisense; D010739:Phospholipase D; D017853:Phosphotransferases (Alcohol Group Acceptor); D017452:Receptors, IgG; D000099157:Sphingosine Kinase; C105606:phospholipase D2; C106801:phospholipase D1