Skip to main content
SpringerLink
Log in

iPLA2 Activation Mediates Granular Exocytosis and Corrects Microbicidal Defects in ROS-Deficient and CGD Human Neutrophils

  • Original Article
  • Published:
Journal of Clinical Immunology Aims and scope Submit manuscript

Abstract

Purpose

The ubiquitous calcium–independent phospholipase A2 enzyme (iPLA2) is inhibited by calmodulin binding and known to be responsible for phospholipid remodeling housekeeping functions including granule exocytosis–associated membrane fusion in normal human neutrophils. We evaluate in human neutrophils the iPLA2 secretagogue effects using normal neutrophils, where reactive oxygen species (ROS) generation has been blocked by diphenyleneiodonium, as well as in neutrophils from chronic granulomatous disease (CGD) patients.

Methods

Neutrophils were pretreated with W7, a calmodulin inhibitor known to activate iPLA2 and exocytosis of granules, and vesicles as well as intra- and extra-microbicidal activity against Staphylococcus aureus and Aspergillus fumigatus were evaluated.

Results

W7 increases exocytosis of primary, secondary, and tertiary granules and vesicles and improves neutrophil microbicidal activity against S. aureus and A. fumigatus.

Conclusions

In neutrophils, calmodulin-mediated iPLA2 inhibition controls granule and vesicle exocytosis in the phagosome and in the extracellular microenvironment. Relieving iPLA2 inhibition results in increased exocytosis of primary, secondary, and tertiary granules and secretory vesicles with correction of defective intracellular and extracellular microbicidal activity. In CGD patients presenting ROS defective production, this increase in the non-oxidative killing pathway partially corrects their microbicidal defects.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

BEL:

bromoenol lactone

Cyt B:

cytochalasin B

CGD:

chronic granulomatose disease

DCFH:

dichloro-dihydro-fluorescein

DPI:

diphenyleneiodonium

iPLA2:

calcium-independent phospholipase A2

MMP-9:

matrix metallo-proteinase 9

MPO:

myeloperoxidase

PAF:

platelet-activating factor

PMN:

polymorphonuclear leukocyte

ROS:

reactive oxygen species

XTT:

sodium 3,3′-[1(phenylamino)carbonyl]-3,4-tetrazolium]-3is(4-methoxy-6-nitro) benzene sulfonic acid hydrate

References

  1. Williams R. Killing controversy. J Exp Med. 2006;203(11):2404.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Nauseef WM. How human neutrophils kill and degrade microbes: an integrated view. Immunol Rev. 2007;219:88–102.

    Article  CAS  PubMed  Google Scholar 

  3. Standish AJ, Weiser JN. Human neutrophils kill Streptococcus pneumoniae via serine proteases. J Immunol. 2009;183(4):2602–9.

    Article  CAS  PubMed  Google Scholar 

  4. Ramanadham S, Ali T, Ashley JW, Bone RN, Hancock WD, Lei X. Calcium-independent phospholipases A2 and their roles in biological processes and diseases. J Lipid Res. 2015;56(9):1643–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Larsson Forsell PK, Runarsson G, Ibrahim M, Björkholm M, Claesson HE. On the expression of cytosolic calcium-independent phospholipase A2 (88kDa) in immature and mature myeloid cells and its role in leukotriene synthesis in human granulocytes. FEBS Lett. 1998;434(3):295–9.

    Article  CAS  PubMed  Google Scholar 

  6. Zhang D, Shooshtarizadeh P, Laventie BJ, Colin DA, Chich JF, Vidic J, et al. Two chromogranin a-derived peptides induce calcium entry in human neutrophils by calmodulin-regulated calcium independent phospholipase A2. PLoS One. 2009;4(2):e4501.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Ellis JA, Mayer SJ, Jones OT. The effect of the NADPH oxidase inhibitor diphenyleneiodonium on aerobic and anaerobic microbicidal activities of human neutrophils. Biochem J. 1988;251(3):887–91.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Simard JC, Girard D, Tessier PA. Induction of neutrophil degranulation by S100A9 via a MAPK-dependent mechanism. J Leukoc Biol. 2010;87(5):905–14.

    Article  CAS  PubMed  Google Scholar 

  9. Stemerding AM, Köhl J, Pandey MK, Kuipers A, Leusen JH, Boross P, et al. Staphylococcus aureus formyl peptide receptor-like 1 inhibitor (FLIPr) and its homologue FLIPr-like are potent FcγR antagonists that inhibit IgG-mediated effector functions. J Immunol. 2013;191(1):353–62.

    Article  CAS  PubMed  Google Scholar 

  10. Flamand L, Tremblay MJ, Borgeat P. Leukotriene B4 triggers the in vitro and in vivo release of potent antimicrobial agents. J Immunol. 2007;178(12):8036–45.

    Article  CAS  PubMed  Google Scholar 

  11. Lee MJ, Liu H, Barker BM, Snarr BD, Gravelat FN, Al Abdallah Q, et al. The fungal exopolysaccharide galactosaminogalactan mediates virulence by enhancing resistance to neutrophil extracellular traps. PLoS Pathog. 2015;11(10):e1005187.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Jog NR, Rane MJ, Lominadze G, Luerman GC, Ward RA, McLeish KR. The actin cytoskeleton regulates exocytosis of all neutrophil granule subsets. Am J Physiol Cell Physiol. 2007;292(5):C1690–700.

    Article  CAS  PubMed  Google Scholar 

  13. Borregaard N, Sørensen OE, Theilgaard-Mönch K. Neutrophil granules: a library of innate immunity proteins. Trends Immunol. 2007;28(8):340–5.

    Article  CAS  PubMed  Google Scholar 

  14. Jian Lian JP, Crossley L, Zhan Q, Huang R, Coffer P, Toker A, et al. Antagonists of calcium fluxes and calmodulin block activation of the p21-activated protein kinases in neutrophils. J Immunol. 2001;166(4):2643–50.

    Article  Google Scholar 

  15. Verploegen S, van Leeuwen CM, van Deutekom HW, Lammers JW, Koenderman L, Coffer PJ. Role of Ca2+/calmodulin regulated signaling pathways in chemoattractant induced neutrophil effector functions. Comparison with the role of phosphotidylinositol-3 kinase. Eur J Biochem. 2002;269(18):4625–34.

    Article  CAS  PubMed  Google Scholar 

  16. Jenkins CM, Han X, Mancuso DJ, Gross RW. Identification of calcium-independent phospholipase A2 (iPLA2) beta, and not iPLA2gamma, as the mediator of arginine vasopressin-induced arachidonic acid release in A-10 smooth muscle cells. Enantioselective mechanism-based discrimination of mammalian iPLA2s. J Biol Chem. 2002;277:32807–14.

    Article  CAS  PubMed  Google Scholar 

  17. Odell EW, Segal AW. Killing of pathogens associated with chronic granulomatous disease by the non-oxidative microbicidal mechanisms of human neutrophils. J Med Microbiol. 1991;34(3):129–35.

    Article  CAS  PubMed  Google Scholar 

  18. Leliefeld PH, Koenderman L, Pillay J. How neutrophils shape adaptive immune responses. Front Immunol. 2015;6:471.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Bakema JE, Ganzevles SH, Fluitsma DM, Schilham MW, Beelen RH, Valerius T, et al. Targeting FcαRI on polymorphonuclear cells induces tumor cell killing through autophagy. J Immunol. 2011;187(2):726–32.

    Article  CAS  PubMed  Google Scholar 

  20. Driessen HE, Bourgonje VJ, van Veen TA, Vos MA. New antiarrhythmic targets to control intracellular calcium handling. Neth Heart J. 2014;22(5):198–213.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Ashpole NM, Song W, Brustovetsky T, Engleman EA, Brustovetsky N, Cummins TR, et al. Calcium/calmodulin-dependent protein kinase II (CaMKII) inhibition induces neurotoxicity via dysregulation of glutamate/calcium signaling and hyperexcitability. J Biol Chem. 2012;287:8495–506.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Nikolaou A, Kokotou MG, Vasilakaki S, Kokotos G. Small-molecule inhibitors as potential therapeutics and as tools to understand the role of phospholipases A2. Biochim Biophys Acta Mol Cell Biol Lipids. 2019 Jun;1864(6):941–56.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

The authors thank Drs. Christophe Chantrain, Alice Ferster, Benoit Florkin, Pierre Philippet, and Jutte Van Der Werff Ten Bosch for the CGD blood samples and Drs. Brigitte Cantiniaux and Francis Corazza for the discussion.

Funding

This work was supported by the Kriibskrank Kanner Foundatioun of Luxembourg.

Author information

Authors and Affiliations

  1. Pediatric Oncology Laboratory, Hôpital Universitaire des Enfants, 1020, Brussels, Belgium

    Issam Harfi, Stéphanie D’Hondt & Eric Sariban

  2. Cancer Unit, Hôpital Universitaire des Enfants, 15, Avenue Jean-Joseph Crocq, 1020, Brussels, Belgium

    Issam Harfi & Eric Sariban

Authors
  1. Issam Harfi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Stéphanie D’Hondt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eric Sariban
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Issam Harfi.

Ethics declarations

PMN were obtained from 9 CGD patients after written authorization in accordance with Declaration of Helsinki, following protocol approved by the Hôpital des Enfants ethical committee

Conflict of Interest

The authors declare that they have no competing interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic Supplementary Material

ESM 1

(DOCX 223 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Harfi, I., D’Hondt, S. & Sariban, E. iPLA2 Activation Mediates Granular Exocytosis and Corrects Microbicidal Defects in ROS-Deficient and CGD Human Neutrophils. J Clin Immunol 39, 486–493 (2019). https://doi.org/10.1007/s10875-019-00630-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10875-019-00630-7

Keywords

Search

Navigation