Exocytosis of neutrophil granule subsets and activation of prolyl isomerase 1 are required for respiratory burst priming.
This study tested the hypothesis that priming the neutrophil respiratory burst requires both granule exocytosis and activation of the prolyl isomerase Pin1. Fusion proteins containing the TAT cell permeability sequence and either the SNARE domain of syntaxin-4 or the N-terminal SNARE domain of SNAP-23 were used to examine the role of granule subsets in TNF-mediated respiratory burst priming using human neutrophils. Concentration-inhibition curves for exocytosis of individual granule subsets and for priming of fMLF-stimulated superoxide release and phagocytosis-stimulated H2O2 production were generated. Maximal inhibition of priming ranged from 72 to 88%. Linear regression lines for inhibition of priming versus inhibition of exocytosis did not differ from the line of identity for secretory vesicles and gelatinase granules, while the slopes or the y-intercepts were different from the line of identity for specific and azurophilic granules. Inhibition of Pin1 reduced priming by 56%, while exocytosis of secretory vesicles and specific granules was not affected. These findings indicate that exocytosis of secretory vesicles and gelatinase granules and activation of Pin1 are independent events required for TNF-mediated priming of neutrophil respiratory burst.
Department of Medicine, University of Louisville, Louisville, KY 40202, USA. k.mcleish@louisville.edu
Kenneth R
KR
McLeish
Silvia M
SM
Uriarte
Shweta
S
Tandon
Timothy M
TM
Creed
Junyi
J
Le
Richard A
RA
Ward
D004789:Enzyme Activation / Q000187:drug effects / Q000276:immunology; D005089:Exocytosis / Q000187:drug effects / Q000276:immunology*; D005260:Female; D006801:Humans; D008297:Male; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology*; D019696:Peptidylprolyl Isomerase / Q000276:immunology* / Q000378:metabolism; D050765:Qa-SNARE Proteins / Q000276:immunology / Q000378:metabolism; D050766:Qb-SNARE Proteins / Q000276:immunology / Q000378:metabolism; D050767:Qc-SNARE Proteins / Q000276:immunology / Q000378:metabolism; D016897:Respiratory Burst / Q000187:drug effects / Q000276:immunology*; D022142:Secretory Vesicles / Q000201:enzymology / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D050765:Qa-SNARE Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; C100493:SNAP23 protein, human; D014409:Tumor Necrosis Factor-alpha; D009240:N-Formylmethionine Leucyl-Phenylalanine; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase