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Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels

Received: 1 June 2020     Accepted: 15 June 2020     Published: 29 June 2020
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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.

Published in International Journal of Microbiology and Biotechnology (Volume 5, Issue 3)
DOI 10.11648/j.ijmb.20200503.16
Page(s) 110-119
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2020. Published by Science Publishing Group

Keywords

Pore-forming Toxin, S. aureus, Leukotoxin, Cl - channels, γ-hemolysin, Panton-Valentin Leukocidin, Neutrophils, Spectrofluorometry

References
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[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.
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[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.
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    Leïla Staali, Didier André Colin. (2020). Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels. International Journal of Microbiology and Biotechnology, 5(3), 110-119. https://doi.org/10.11648/j.ijmb.20200503.16

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    Leïla Staali; Didier André Colin. Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels. Int. J. Microbiol. Biotechnol. 2020, 5(3), 110-119. doi: 10.11648/j.ijmb.20200503.16

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    AMA Style

    Leïla Staali, Didier André Colin. Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels. Int J Microbiol Biotechnol. 2020;5(3):110-119. doi: 10.11648/j.ijmb.20200503.16

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  • @article{10.11648/j.ijmb.20200503.16,
      author = {Leïla Staali and Didier André Colin},
      title = {Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels},
      journal = {International Journal of Microbiology and Biotechnology},
      volume = {5},
      number = {3},
      pages = {110-119},
      doi = {10.11648/j.ijmb.20200503.16},
      url = {https://doi.org/10.11648/j.ijmb.20200503.16},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20200503.16},
      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.},
     year = {2020}
    }
    

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  • TY  - JOUR
    T1  - Bi-component Staphylococcal Leukotoxins Induce Chloride Ions Fluxes in Human Neutrophils: Opening of Ca2+-activated Cl- Channels
    AU  - Leïla Staali
    AU  - Didier André Colin
    Y1  - 2020/06/29
    PY  - 2020
    N1  - https://doi.org/10.11648/j.ijmb.20200503.16
    DO  - 10.11648/j.ijmb.20200503.16
    T2  - International Journal of Microbiology and Biotechnology
    JF  - International Journal of Microbiology and Biotechnology
    JO  - International Journal of Microbiology and Biotechnology
    SP  - 110
    EP  - 119
    PB  - Science Publishing Group
    SN  - 2578-9686
    UR  - https://doi.org/10.11648/j.ijmb.20200503.16
    AB  - 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.
    VL  - 5
    IS  - 3
    ER  - 

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Author Information
  • Bacteriology Institute of Medical Faculty, Louis Pasteur University, Strasbourg, France

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