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An in vitro α-neurotoxin-nAChR binding assay correlates with lethality and in vivo neutralization of a large number of elapid neurotoxic snake venoms from four continents.
- Source :
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PLoS neglected tropical diseases [PLoS Negl Trop Dis] 2020 Aug 28; Vol. 14 (8), pp. e0008581. Date of Electronic Publication: 2020 Aug 28 (Print Publication: 2020). - Publication Year :
- 2020
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Abstract
- The aim of this study was to develop an in vitro assay for use in place of in vivo assays of snake venom lethality and antivenom neutralizing potency. A novel in vitro assay has been developed based on the binding of post-synaptically acting α-neurotoxins to nicotinic acetylcholine receptor (nAChR), and the ability of antivenoms to prevent this binding. The assay gave high correlation in previous studies with the in vivo murine lethality tests (Median Lethal Dose, LD50), and the neutralization of lethality assays (Median Effective Dose, ED50) by antisera against Naja kaouthia, Naja naja and Bungarus candidus venoms. Here we show that, for the neurotoxic venoms of 20 elapid snake species from eight genera and four continents, the in vitro median inhibitory concentrations (IC50s) for α-neurotoxin binding to purified nAChR correlated well with the in vivo LD50s of the venoms (R2 = 0.8526, p < 0.001). Furthermore, using this assay, the in vitro ED50s of a horse pan-specific antiserum against these venoms correlated significantly with the corresponding in vivo murine ED50s, with R2 = 0.6896 (p < 0.01). In the case of four elapid venoms devoid or having a very low concentration of α-neurotoxins, no inhibition of nAChR binding was observed. Within the philosophy of 3Rs (Replacement, Reduction and Refinement) in animal testing, the in vitro α-neurotoxin-nAChR binding assay can effectively substitute the mouse lethality test for toxicity and antivenom potency evaluation for neurotoxic venoms in which α-neurotoxins predominate. This will greatly reduce the number of mice used in toxicological research and antivenom production laboratories. The simpler, faster, cheaper and less variable in vitro assay should also expedite the development of pan-specific antivenoms against various medically important snakes in many parts of the world.<br />Competing Interests: The authors have declared that no competing interests exist.
- Subjects :
- Africa
Americas
Animals
Asia
Australia
Elapid Venoms immunology
Elapid Venoms toxicity
Elapidae immunology
Horses
Humans
Immune Sera immunology
Mice
Neurotoxins immunology
Neurotoxins toxicity
Neutralization Tests
Snake Bites immunology
Snake Bites mortality
Biological Assay methods
Elapid Venoms chemistry
Neurotoxins chemistry
Receptors, Nicotinic chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1935-2735
- Volume :
- 14
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- PLoS neglected tropical diseases
- Publication Type :
- Academic Journal
- Accession number :
- 32857757
- Full Text :
- https://doi.org/10.1371/journal.pntd.0008581