A cell-based assay for screening of antidotes to, and antivenom against Chironex fleckeri (box jellyfish) venom

N KONSTANTAKOPOULOS, Geoffrey Isbister, J SEYMOUR, W HODGSON

    Research output: Contribution to journalArticleResearchpeer-review

    Abstract

    Introduction: Chironex fleckeri is a large box jellyfish that has been labelled the 'most venomous animal' in the world. We have recently shown that the primary effect of C. fleckeri venom in vivo is cardiovascular collapse. This study utilised a cell-based assay to examine the effects of C. fleckeri venom on the proliferation of a rat aortic smooth muscle cell line. In addition, the ability of CSL box jellyfish antivenom and/or various potential treatment strategies to neutralise the effects of the venom was examined. Methods: A7r5 cells were cultured in media containing venom. The effect of CSL box jellyfish antivenom (5�U/mL), CSL polyvalent snake antivenom (5�U/mL), lanthanum (5�?M), MgSO4 (50�mM), verapamil (5�?M) or felodipine (5�?M) was examined. Cell viability was determined using a Cell titer 96 AQueous One Solution cell proliferation assay. Results: Incubation of A7r5 cells with serially diluted venom (2-0.004�?g/mL) caused a concentration-dependent inhibition of cell proliferation with an IC50 value of 0.056�?g/mL. This response was not affected by the absence of calcium or the presence of lanthanum in the media. Box jellyfish antivenom (5�U/mL) prevented the inhibition of cell proliferation caused by the venom. Verapamil (5�?M) had no significant effect on the inhibition. In contrast, felodipine (5�?M) or MgSO4 (50�mM) potentiated the effects of the venom and partially negated the protective effect of the antivenom. Discussion: This study displayed the ability to utilise a cell-based assay to determine the effects of C. fleckeri venom on vascular cell viability. It showed that CSL box jellyfish can neutralise the effects of the venom but only if added prior to the venom. In addition, potential adjunct therapies verapamil, felodipine and MgSO4 were found to be ineffective, with felodipine and MgSO4 potentiating the detrimental effects of the venom. � 2009 Elsevier Inc. All rights reserved.
    Original languageEnglish
    Pages (from-to)166-170
    Number of pages5
    JournalJournal of Pharmacological and Toxicological Methods
    Volume59
    Issue number3
    Publication statusPublished - 2009

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    Cnidarian Venoms
    Cubozoa
    Antivenins
    Antidotes
    Venoms
    Assays
    Screening
    Felodipine
    Cell proliferation
    Verapamil
    Lanthanum
    Cells
    Cell Proliferation
    Cell Survival
    Snakes

    Cite this

    KONSTANTAKOPOULOS, N ; Isbister, Geoffrey ; SEYMOUR, J ; HODGSON, W. / A cell-based assay for screening of antidotes to, and antivenom against Chironex fleckeri (box jellyfish) venom. In: Journal of Pharmacological and Toxicological Methods. 2009 ; Vol. 59, No. 3. pp. 166-170.
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    title = "A cell-based assay for screening of antidotes to, and antivenom against Chironex fleckeri (box jellyfish) venom",
    abstract = "Introduction: Chironex fleckeri is a large box jellyfish that has been labelled the 'most venomous animal' in the world. We have recently shown that the primary effect of C. fleckeri venom in vivo is cardiovascular collapse. This study utilised a cell-based assay to examine the effects of C. fleckeri venom on the proliferation of a rat aortic smooth muscle cell line. In addition, the ability of CSL box jellyfish antivenom and/or various potential treatment strategies to neutralise the effects of the venom was examined. Methods: A7r5 cells were cultured in media containing venom. The effect of CSL box jellyfish antivenom (5�U/mL), CSL polyvalent snake antivenom (5�U/mL), lanthanum (5�?M), MgSO4 (50�mM), verapamil (5�?M) or felodipine (5�?M) was examined. Cell viability was determined using a Cell titer 96 AQueous One Solution cell proliferation assay. Results: Incubation of A7r5 cells with serially diluted venom (2-0.004�?g/mL) caused a concentration-dependent inhibition of cell proliferation with an IC50 value of 0.056�?g/mL. This response was not affected by the absence of calcium or the presence of lanthanum in the media. Box jellyfish antivenom (5�U/mL) prevented the inhibition of cell proliferation caused by the venom. Verapamil (5�?M) had no significant effect on the inhibition. In contrast, felodipine (5�?M) or MgSO4 (50�mM) potentiated the effects of the venom and partially negated the protective effect of the antivenom. Discussion: This study displayed the ability to utilise a cell-based assay to determine the effects of C. fleckeri venom on vascular cell viability. It showed that CSL box jellyfish can neutralise the effects of the venom but only if added prior to the venom. In addition, potential adjunct therapies verapamil, felodipine and MgSO4 were found to be ineffective, with felodipine and MgSO4 potentiating the detrimental effects of the venom. � 2009 Elsevier Inc. All rights reserved.",
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    author = "N KONSTANTAKOPOULOS and Geoffrey Isbister and J SEYMOUR and W HODGSON",
    year = "2009",
    language = "English",
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    A cell-based assay for screening of antidotes to, and antivenom against Chironex fleckeri (box jellyfish) venom. / KONSTANTAKOPOULOS, N; Isbister, Geoffrey; SEYMOUR, J; HODGSON, W.

    In: Journal of Pharmacological and Toxicological Methods, Vol. 59, No. 3, 2009, p. 166-170.

    Research output: Contribution to journalArticleResearchpeer-review

    TY - JOUR

    T1 - A cell-based assay for screening of antidotes to, and antivenom against Chironex fleckeri (box jellyfish) venom

    AU - KONSTANTAKOPOULOS, N

    AU - Isbister, Geoffrey

    AU - SEYMOUR, J

    AU - HODGSON, W

    PY - 2009

    Y1 - 2009

    N2 - Introduction: Chironex fleckeri is a large box jellyfish that has been labelled the 'most venomous animal' in the world. We have recently shown that the primary effect of C. fleckeri venom in vivo is cardiovascular collapse. This study utilised a cell-based assay to examine the effects of C. fleckeri venom on the proliferation of a rat aortic smooth muscle cell line. In addition, the ability of CSL box jellyfish antivenom and/or various potential treatment strategies to neutralise the effects of the venom was examined. Methods: A7r5 cells were cultured in media containing venom. The effect of CSL box jellyfish antivenom (5�U/mL), CSL polyvalent snake antivenom (5�U/mL), lanthanum (5�?M), MgSO4 (50�mM), verapamil (5�?M) or felodipine (5�?M) was examined. Cell viability was determined using a Cell titer 96 AQueous One Solution cell proliferation assay. Results: Incubation of A7r5 cells with serially diluted venom (2-0.004�?g/mL) caused a concentration-dependent inhibition of cell proliferation with an IC50 value of 0.056�?g/mL. This response was not affected by the absence of calcium or the presence of lanthanum in the media. Box jellyfish antivenom (5�U/mL) prevented the inhibition of cell proliferation caused by the venom. Verapamil (5�?M) had no significant effect on the inhibition. In contrast, felodipine (5�?M) or MgSO4 (50�mM) potentiated the effects of the venom and partially negated the protective effect of the antivenom. Discussion: This study displayed the ability to utilise a cell-based assay to determine the effects of C. fleckeri venom on vascular cell viability. It showed that CSL box jellyfish can neutralise the effects of the venom but only if added prior to the venom. In addition, potential adjunct therapies verapamil, felodipine and MgSO4 were found to be ineffective, with felodipine and MgSO4 potentiating the detrimental effects of the venom. � 2009 Elsevier Inc. All rights reserved.

    AB - Introduction: Chironex fleckeri is a large box jellyfish that has been labelled the 'most venomous animal' in the world. We have recently shown that the primary effect of C. fleckeri venom in vivo is cardiovascular collapse. This study utilised a cell-based assay to examine the effects of C. fleckeri venom on the proliferation of a rat aortic smooth muscle cell line. In addition, the ability of CSL box jellyfish antivenom and/or various potential treatment strategies to neutralise the effects of the venom was examined. Methods: A7r5 cells were cultured in media containing venom. The effect of CSL box jellyfish antivenom (5�U/mL), CSL polyvalent snake antivenom (5�U/mL), lanthanum (5�?M), MgSO4 (50�mM), verapamil (5�?M) or felodipine (5�?M) was examined. Cell viability was determined using a Cell titer 96 AQueous One Solution cell proliferation assay. Results: Incubation of A7r5 cells with serially diluted venom (2-0.004�?g/mL) caused a concentration-dependent inhibition of cell proliferation with an IC50 value of 0.056�?g/mL. This response was not affected by the absence of calcium or the presence of lanthanum in the media. Box jellyfish antivenom (5�U/mL) prevented the inhibition of cell proliferation caused by the venom. Verapamil (5�?M) had no significant effect on the inhibition. In contrast, felodipine (5�?M) or MgSO4 (50�mM) potentiated the effects of the venom and partially negated the protective effect of the antivenom. Discussion: This study displayed the ability to utilise a cell-based assay to determine the effects of C. fleckeri venom on vascular cell viability. It showed that CSL box jellyfish can neutralise the effects of the venom but only if added prior to the venom. In addition, potential adjunct therapies verapamil, felodipine and MgSO4 were found to be ineffective, with felodipine and MgSO4 potentiating the detrimental effects of the venom. � 2009 Elsevier Inc. All rights reserved.

    KW - antidote

    KW - calcium

    KW - felodipine

    KW - jellyfish antivenom

    KW - lanthanum

    KW - magnesium sulfate

    KW - unclassified drug

    KW - venom

    KW - verapamil

    KW - animal cell

    KW - animal experiment

    KW - animal model

    KW - animal tissue

    KW - article

    KW - cell based assay

    KW - cell proliferation

    KW - cell viability

    KW - concentration response

    KW - controlled study

    KW - IC 50

    KW - in vivo study

    KW - jellyfish

    KW - nonhuman

    KW - rat

    KW - toxin analysis

    KW - Animals

    KW - Antidotes

    KW - Antivenins

    KW - Aorta

    KW - Biological Assay

    KW - Cell Line

    KW - Cell Proliferation

    KW - Cell Survival

    KW - Cnidarian Venoms

    KW - Cubozoa

    KW - Felodipine

    KW - Magnesium Sulfate

    KW - Muscle, Smooth, Vascular

    KW - Myocytes, Smooth Muscle

    KW - Rats

    KW - Verapamil

    KW - Animalia

    KW - Chironex fleckeri

    KW - Rattus

    M3 - Article

    VL - 59

    SP - 166

    EP - 170

    JO - Journal of Pharmacological and Toxicological Methods

    JF - Journal of Pharmacological and Toxicological Methods

    SN - 1056-8719

    IS - 3

    ER -