Volume 5, Issue 3, September 2020, Page: 110-119
Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels
Leïla Staali, Department of Biotechnology, Natural and Life Sciences Faculty, Ahmed Ben Bella Oran1-University, Oran, Algeria; Bacteriology Institute of Medical Faculty, Louis Pasteur University, Strasbourg, France
Didier André Colin, Bacteriology Institute of Medical Faculty, Louis Pasteur University, Strasbourg, France
Received: Jun. 1, 2020;       Accepted: Jun. 15, 2020;       Published: Jun. 29, 2020
DOI: 10.11648/j.ijmb.20200503.16      View  81      Downloads  31
Abstract
The bi-component leukotoxins; γ-hemolysin and Panton and Valentin Leukocidin (PVL) from Staphylococcus aureus induce two independent cellular events 1) the formation of trans-membrane pores not permeable to chloride (Cl-) ions and 2) the activation of at least, two modes of chloride fluxes (efflux/influx), including pre-existing Ca2+-activated Cl- channels (CaCC) in human polymorphonuclear neutrophils (PMNs). This was investigated by using spectrofluorometry techniques and the chloride-sensitive quencher fluorescent indicator, MQAE (N-(6-methoxyquinolyl) acetoacethyl ester). The ethidium bromide was used as an indicator for the trans-membrane pores formation by staphylococcal leukotoxins. In the absence of extracellular Ca2+, HlgA/HlgB, HlgC/HlgB and LukS-PV/LukF-PV leukotoxins from S. aureus induced a massive efflux of chloride (Cl-) ions. Interestingly, in the presence of extracellular Ca2+, the HlgA/HlgB γ-hemolysin provoked a biphasic response of Cl- movements (efflux/influx). Conversely to HlgA/HlgB and LukS-PV/LukF-PV, HlgC/HlgB leukotoxins did not induce any Cl- movement under this condition (e.g. in the presence of extracellular Ca2+). The potent Cl- channel inhibitor, DIDS, did inhibit significantly the Cl- fluxes caused by all pairs of staphylococcal leukotoxins tested in both conditions. In the present study, we found that the inhibitory effect of flufenamic acid, known as a Cl- channel inhibitor, was restricted only to the Ca2+-dependent Cl- influx triggered only by HlgA/HlgB and LukS-PV/LukF-PV leukotoxins. These findings might suggest that, Cl- fluxes in human neutrophils did involve at least, two different types of Cl- pathways, depending on the absence or presence of extracellular Ca2+. Both Cl- channels blockers, DIDS and flufenamic acid did not alter the pores formation by staphylococcal leukotoxins. Furthermore, under conditions when the membrane pores formation was blocked by divalent ions (Ca2+ and/or Zn2+), Cl- ions movements were still observed. Taken together, our results strongly provide an evidence that: i) trans-membrane pores formed by staphylococcal leukotoxins: HlgA/HlgB, HlgC/HlgB (γ-hemolysin) and LukS-PV/LukF-PV (PVL) do not drive Cl- ions fluxes ii) at least, two different types of Cl- ions pathways are activated, depending on the absence or presence of extracellular Ca2+, including Ca2+-activated Cl- channels (CaCC) and, iii) Ca2+-activated Cl- channels are mediated only by HlgA/HlgB and LukS-PV/LukF-PV leukotoxins.
Keywords
Pore-forming Toxin, S. aureus, Leukotoxin, Cl - channels, γ-hemolysin, Panton-Valentin Leukocidin, Neutrophils, Spectrofluorometry
To cite this article
Leïla Staali, Didier André Colin, Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels, International Journal of Microbiology and Biotechnology. Vol. 5, No. 3, 2020, pp. 110-119. doi: 10.11648/j.ijmb.20200503.16
Copyright
Copyright © 2020 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Prévost G, Cribier B, Couppié P, Petiau P, Supersac G, Finck-Barbancon V, Monteil H, Piémont Y (1995) Panton-Valentine leucocidin and gamma-hemolysin from Staphylococcus aureus ATCC49775 are encoded by distinct loci and have different biological activities. Infect. Immun. 63: 4121-4129.
[2]
Finck-Barbancon V, Prévost G, Piémont Y (1991) Improved purification of leukocidin from Staphylococcus aureus and toxin distribution among hospital strains. Res. Microbiol. 142: 75-85.
[3]
Cribier B, Prévost G, Couppié P, Finck-Barbancon V, Grosshans E, Piémont Y (1992) Staphylococcus aureus leukocidin: a new virulence factor in cutaneous infections? An epidemiological and experimental study. Dermatology 185: 175-180.
[4]
Couppié P, Cribier B, Prévost G, Grosshans E, Piemont Y (1994) Leukocidin from Staphylococcus aureus and cutaneous infections: an epidemiological study. Arch. Dermatol. (130): 1208-1209.
[5]
Colin DA, Mazurier I, Sire S, Finck-Barbançon V (1994) Interaction of the two components of leukocicin from Staphylococcus aureus with human polymorphonuclear leukocyte membranes: sequenctiel binding and subsequent activation. Infect. Immun. 62: 3184-3188.
[6]
Guaduchon V, Werner S, Prévost G, Monteil H, Colin DA (2001) Flow cytometry determination of Panton-Valentin leukocidin S component binding. Infect. Immun. 69: 2390-2395.
[7]
Staali L, Monteil H, Colin DA (1998) The pore-forming leukotoxins from Staphylococcus aureus open Ca2+ channels in human polymorphonuclear neutrophils. J. Membr. Biol. 162: 209-216.
[8]
Finck-Barbançon V, Duportail G, Meunier O, Colin DA (1993) Pore formation by a two-component leukocidin from Staphylococcus aureus within the membrane of human polymorphonuclear leukocytes. Biochim. Biophys. Acta. 1128: 275-282.
[9]
Ferreras M, Hoper F, Dalla Serra M, Colin DA, Prévost G, Menestrina G (1998) The interaction of Staphylococcus aureus bi-component γ-hemolysin and leucocidins with cells and lipid membranes. Biochim. Biophys. Acta 1414: 108-126.
[10]
Pedelacq JD, Maveyraud L, Prevost G, Baba-Moussa L, Gonzalez A, Courcelle E, Shepard W, Monteil H, Samama JP, Mourey L (1999) The structure of Staphylococcus aureus leucocidin component (LukF-PV) reveals the fold of the water-soluble species of a family of transmembrane pore-forming toxins. Structure 7: 277-287.
[11]
Staali L, Monteil H, Colin DA (1998) Action mechanism of staphylococcal bi-component leucotoxins. Toxicon 36 (12): 1746.
[12]
Staali L, Colin DA (2020) The pore-forming leukotoxins from S. aureus involve Ca2+-activated Ca2+ channels and other types of Ca2+ channels in Ca2+ entry into neutrophils. International Journal of Microbiology and Biotechnology. 5 (2): 55-68.
[13]
Colin DA, Monteil H (2003) Control of the oxidative burst of human neutrophils by staphylococcal leukotoxins. Infection and Immunity. 71 (7): 3724-3729.
[14]
Krause KH, Welsh MJ (1990) Voltage-dependent and Ca2+-activated ion channels in human neutrophils. J. Clin. Invest. 85: 491-498.
[15]
Schumann L, Gardner P, Raffin TA (1993) Recombinant human tumor necrosis factor α induces calcium oscillation and calcium-activated chloride current inn human neutrophils. J. Biol. Chem. 268: 2134-2140.
[16]
Schumann MA, Raffin TA (1994) Activation of voltage-dependent chloride current in human neutrophils by phorbol 12-myristate 13-acetate and formyl-methionyl-leucyl-phenylalanine. The role of protein kinase. J. Biol. Chem. 269: 2389-2398.
[17]
Simchowitz L, Textor J, Cragoe EJ (1993) Cell volume regulation in human neutrophils: 2-(aminoethyl) phenols as Cl- channel inhibitors. Am. J. Physiol. 265: C143-155.
[18]
Gallin EK (1991) Ion channels in leukocytes. Physiol. Rev. 3: 775-811.
[19]
Koncz C, Daugirdas J (1994) Use of MQAE for measurement of intracellular [Cl-] in cultured aortic smouth muscle cells. Am. J. Physiol. 267: H2114-2123.
[20]
Ikeuchi Y, Kogiso H, Hosogi S, Tanaka S, Shimmamoto C, Inui T, Nakahari T, Marunaka Y (2018) Measurement of [Cl-] unaffected by the cell volume change using MQAE-based two-photon microscopy in airway ciliary cells of mice. The Journal of Physiological Sciences 68: 191-199.
[21]
Shimizu Y, Daniels RH, Elmore MA, Finnen MJ, Hill ME, Lackie JM (1993) Agonist-stimuated Cl- efflux from human neutrophils: a common phenomenon during neutrophil activation. Biochem. Pharmacol. 45: 1743-1751.
[22]
Menegazzi R, Busetto S, Dri P, Cramer R, Patriarca P (1996) Chloride ion efflux regulates adherence, spreading, and respiratory burst of neutrophils stimulated by tumor necrosis factor-α (TNF) on biologic surfaces J. Cell Biol. 135: 511-522.
[23]
Geddes CD, Apperson K, Karolin J, Birch DJ (2001) Chloride-sensitive fluorescent indicators. Anal. Biochem. 293 (1): 60-66.
[24]
West MR, Molloy CR (1996) A microplate assay measuring chloride ion channel activity. Anal. Biochem. 241: 51-58.
[25]
Phipps DJ, Branch DR, Schlichter LC (1996) Chloride channel blockers inhibits T lymphocytes activation and signaling. Cell Signal 8: 141-149.
[26]
Greenwood I, Large W (1995) Comparison of the effects of fenamates on Ca2+-activated chloride and potassium currents in rabbit portal vein smouth muscle cells. Br. J. Pharmacol. 116: 2939-2948.
[27]
Busetto S, Trevisan E, Decleva E, Dri P, Meneqazzi R (2007) Chloride movements in human neutrophils during phagocytosis: characterization and relationship to granule release. The journal of Immunology 179: 4110-4124.
[28]
Menegazzi R, Busetto S, Decleva E, Cramer R, Dri P, Patriarca P (1999) Triggering of chloride ion efflux from human neutrophils as a novel function of leukocyte β2 intergrins: relationship with spreading and activation of the respiratory burst. J. immunol. 162: 423-434.
[29]
Engblom AC, Akerman KE (1993) Determination of the intracellular free chloride concentration in rat brain synaptoneurosomes using a chloride-sensitive fluorescent indicator. Biochem. Biophys. Acta 1153 (2): 262-266.
[30]
Kanno T, Takishima T (1990) Chloride and potassium channel in U937 human monocytes. J. Membr. Biol. 116: 149-161.
[31]
Simchowitz L (1986) Chloride movements in human neutrophils: diffusion, exchange and active transport. J. Gen. Phys. 88: 167.
[32]
Simchowitz L (1990) Special topic: ion movements in leukocytes. Annu. Rev. Physiol. 52: 363-364.
[33]
Simchowitz L, Bibb JA (1990) Functional analysis of the modes of anion transport in neutrophils and HL-60 cells. Annu. Rev. Physiol. 52: 381-397.
[34]
Di Fulvio M, Aguilar-Bryan L (2019) Chloride transporters and channels in β-cell physiology: revisiting a 40-year-old model. Biochemical Society Transactions 47: 1843-1855.
[35]
Chao AC, Dix JA, Sellers MC, Verkman AS (1999) Fluorescence measurement of chloride transport in monolayer cultured cells. Biophys. J. 56: 1071-1081.
[36]
Kankaanranta H, Moilanen E (1995) Flufenamic and tolfenamic acids inhibit calcium influx in human polymorphonuclear leukocytes. Mol. Pharmacol. 47: 1006-1013.
[37]
Eberhardson M, Patterson S, Grapengiesser E (2000) Microfluorometric analysis of Cl- permeability ad its relation to oxcillatory Ca2+ signaling in glucose-stimulated pancreatic β-cells. Cellular Signalling 12 (11-12): 781-786.
[38]
Andersson C, Roomans GM (2002) Determination of chloride efflux by X-ray microanalysis versus MQAE-fluorescence. Microsc. Res. Tech. 59 (6): 531-532.
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