scholarly journals Neutrophil activation by anti-proteinase 3 antibodies in Wegener's granulomatosis: role of exogenous arachidonic acid and leukotriene B4 generation.

1996 ◽  
Vol 184 (4) ◽  
pp. 1567-1572 ◽  
Author(s):  
F Grimminger ◽  
K Hattar ◽  
C Papavassilis ◽  
B Temmesfeld ◽  
E Csernok ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Wegener’S Granulomatosis ◽  
Wegener's Granulomatosis ◽  
Neutrophil Activation ◽  
Tnf Alpha ◽  
Lipid Mediator ◽  
Human Neutrophils ◽  
Proteinase 3 ◽  
Lipoxygenase Pathway

Among the anti-neutrophil cytoplasmic antibodies (ANCA), those targeting proteinase 3 (PR3) have a high specificity for Wegener's granulomatosis (WG). It is known that a preceding priming of neutrophils with cytokines is a prerequisite for membrane surface expression of PR3, which is then accessible to autoantibody binding. Employing a monoclonal antibody directed against human PR3 and ANCA-positive serum from WG patients with specificity for PR3, we now investigated the role of free arachidonic acid (AA) in autoantibody-related human neutrophil activation. Priming of neutrophils with tumor necrosis factor (TNF-alpha) for 15 min or exposure to anti-PR3 antibodies or incubation with free AA (10 microM) as sole events did not provoke superoxide generation, elastase secretion or generation of 5-lipoxygenase products of AA. Similarly, the combination of TNF-alpha-priming and AA incubation was ineffective. When TNF-alpha-primed neutrophils were stimulated by anti-PR3 antibodies, superoxide and elastase secretion was provoked in the absence of lipid mediator generation. However, when free AA was additionally provided, a strong activation of the 5-lipoxygenase pathway was demasked, with the appearance of excessive quantities of leukotriene (LT)B4, LTA4, and 5-hydroxyeicosatetraenoic acid. Moreover, superoxide and elastase secretion were markedly amplified, and studies with 5-lipoxygenase inhibitors and a LTB4-antagonist demonstrated this was due to an LTB4-related autocrine loop of cell activation. In contrast, the increased synthesis of platelet-activating factor in response to TNF-alpha-priming and anti-PR3 stimulation did not contribute to the amplification loop of neutrophil activation under the given conditions. We conclude that anti-PR3 antibodies are potent inductors of the 5-lipoxygenase pathway in primed human neutrophils, and extracellular free AA, as provided at an inflammatory focus, synergizes with the autoantibodies to evoke full-blown lipid mediator generation, granule secretion and respiratory burst. Such events may be enrolled in the pathogenesis of focal necrotizing vascular injury in Wegener's granulomatosis.

A substrate‐based approach to convert SerpinB1 into a specific inhibitor of proteinase 3, the Wegener's granulomatosis autoantigen

2011 ◽  
Vol 25 (9) ◽  
pp. 3019-3031 ◽  
Author(s):  
Gwenhael Jégot ◽  
Chrystelle Derache ◽  
Sandrine Castella ◽  
Hichem Lahouassa ◽  
Elodie Pitois ◽  
...  
Keyword(s):  
Wegener’S Granulomatosis ◽  
Specific Inhibitor ◽  
Wegener's Granulomatosis ◽  
Proteinase 3

scholarly journals Lipopolysaccharides prime whole human blood and isolated neutrophils for the increased synthesis of 5-lipoxygenase products by enhancing arachidonic acid availability: involvement of the CD14 antigen.

1993 ◽  
Vol 178 (4) ◽  
pp. 1347-1355 ◽  
Author(s):  
M E Surette ◽  
R Palmantier ◽  
J Gosselin ◽  
P Borgeat
Keyword(s):  
Arachidonic Acid ◽  
Mononuclear Cells ◽  
Leukotriene B4 ◽  
Eicosatetraenoic Acid ◽  
Tnf Alpha ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Factor Alpha ◽  
Priming Activity

Stimulation of heparinized blood with 1 microM formyl-methionyl-leucyl-phenylalanine (FMLP) resulted in the formation of < 30 pmol/ml plasma of 5-lipoxygenase (5-LO) products. The preincubation of blood with 1 microgram/ml of lipopolysaccharide (LPS) (Escherichia coli 0111-B4) for 30 min before stimulation with FMLP resulted in the accumulation of 250-300 pmol of 5-LO products per ml plasma. The major products detected were leukotriene B4 and (5S)-hydroxy-6,8,11,14-eicosatetraenoic acid which were produced in equivalent amounts. The priming activity was detectable with as little as 1-10 ng LPS per ml blood and was optimal using 1-10 micrograms LPS/ml blood. The priming for 5-LO product synthesis was optimal after 20-30 min of preincubation with LPS and declined at preincubation times > 30 min. The priming effect of LPS was also observed using the complement fragment C5a or interleukin 8 as agonists. Polymorphonuclear leukocytes (PMN) and peripheral blood mononuclear cells accounted for 80 and 20% of the synthesis of 5-LO products, respectively. The ability of LPS to prime isolated PMN was dependent on the presence of plasma and was inhibited by the anti-CD14 antibody IOM2, indicating a CD14-dependent priming mechanism. The priming of whole blood with tumor necrosis factor alpha (TNF-alpha) and LPS was additive and the presence of mononuclear cells did not enhance the ability of LPS to prime PMN, indicating that the priming activity of LPS is independent of LPS-induced TNF-alpha synthesis. The mechanism by which LPS enhance 5-LO product synthesis in PMN was investigated. Treatment of PMN with LPS strongly enhanced the release of arachidonic acid after stimulation with FMLP. The release of arachidonic acid was optimal 2-3 min after stimulation with FMLP, attaining levels 5-15-fold greater than those observed in unprimed cells stimulated with FMLP. These results demonstrate that LPS dramatically increases the ability of blood to generate 5-LO products, and support the putative role of leukotrienes in pathological states involving LPS.

scholarly journals Neutrophil activation in vitro and in vivo in Wegener's granulomatosis

1994 ◽  
Vol 45 (4) ◽  
pp. 1120-1131 ◽  
Author(s):  
Elisabeth Brouwer ◽  
Minke G. Huitema ◽  
A.H. Leontine Mulder ◽  
Peter Heeringa ◽  
Harry van Goor ◽  
...  
Keyword(s):  
Wegener’S Granulomatosis ◽  
Wegener's Granulomatosis ◽  
Neutrophil Activation

Wegener's Granulomatosis and Antineutrophil Cytoplasmic Autoantibodies

1993 ◽  
Vol 102 (11) ◽  
pp. 906-908 ◽  
Author(s):  
John G. Batsakis ◽  
Adel K. El-Naggar
Keyword(s):  
Laboratory Test ◽  
Wegener’S Granulomatosis ◽  
Wegener's Granulomatosis ◽  
Proteinase 3 ◽  
Inflammatory Disorders ◽  
Serologic Tests ◽  
Criteria For Diagnosis ◽  
Antineutrophil Cytoplasmic Autoantibodies

Serologic tests for antineutrophil cytoplasmic autoantibodies can serve as markers for a number of necrotizing vasculitides and other inflammatory disorders. In the case of Wegener's granulomatosis, an immunofluorescent cytoplasmic pattern and immunoassay for proteinase 3 are quite specific for the disease and are capable of serving as additional criteria for diagnosis. As with any laboratory test, however, results are to be interpreted in the whole clinical and pathologic framework of the disease.

scholarly journals Adenosine Triphosphate Neutralizes Pneumolysin-Induced Neutrophil Activation

2020 ◽  
Vol 222 (10) ◽  
pp. 1702-1712 ◽  
Author(s):  
Fabian Cuypers ◽  
Björn Klabunde ◽  
Manuela Gesell Salazar ◽  
Surabhi Surabhi ◽  
Sebastian B Skorka ◽  
...  
Keyword(s):  
Adenosine Triphosphate ◽  
Extracellular Space ◽  
Cytolytic Activity ◽  
Neutrophil Activation ◽  
Human Neutrophils ◽  
Pneumococcal Infections ◽  
Pneumococcal Strain ◽  
Neutrophil Degranulation ◽  
Neutrophil Response

Abstract Background In tissue infections, adenosine triphosphate (ATP) is released into extracellular space and contributes to purinergic chemotaxis. Neutrophils are important players in bacterial clearance and are recruited to the site of tissue infections. Pneumococcal infections can lead to uncontrolled hyperinflammation of the tissue along with substantial tissue damage through excessive neutrophil activation and uncontrolled granule release. We aimed to investigate the role of ATP in neutrophil response to pneumococcal infections. Methods Primary human neutrophils were exposed to the pneumococcal strain TIGR4 and its pneumolysin-deficient mutant or directly to different concentrations of recombinant pneumolysin. Neutrophil activation was assessed by measurement of secreted azurophilic granule protein resistin and profiling of the secretome, using mass spectrometry. Results Pneumococci are potent inducers of neutrophil degranulation. Pneumolysin was identified as a major trigger of neutrophil activation. This process is partially lysis independent and inhibited by ATP. Pneumolysin and ATP interact with each other in the extracellular space leading to reduced neutrophil activation. Proteome analyses of the neutrophil secretome confirmed that ATP inhibits pneumolysin-dependent neutrophil activation. Conclusions Our findings suggest that despite its cytolytic activity, pneumolysin serves as a potent neutrophil activating factor. Extracellular ATP mitigates pneumolysin-induced neutrophil activation.

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