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Journal of proteomics2017; 158; 62-72; doi: 10.1016/j.jprot.2017.02.015

Proteomic, toxicological and immunogenic characterization of Mexican west-coast rattlesnake (Crotalus basiliscus) venom and its immunological relatedness with the venom of Central American rattlesnake (Crotalus simus).

Abstract: The venom of the Mexican west-coast rattlesnake (Crotalus basiliscus) was characterized for its protein composition, toxicological profile and immunogenic properties. This venom is composed of 68% Zn2+-dependent metalloproteinases (SVMPs), 14% phospholipases A2 (PLA2s), 11% serine proteinases, 4% SVMPs-inhibitor tripeptides (SVMP-ITs), 2% bradykinin-potentiating peptides (BPPs), 0.6% cysteine-rich secretory proteins (CRISPs), and 0.2% l-amino acid oxidases (LAAOs). SVMPs present in the venom are responsible for azocasein hydrolysis and hemorrhagic activity, but their contribution to the lethal activity of the venom in mice is masked by the neurotoxic activity of PLA2s, which in addition are also responsible for myotoxic activity. Despite its relatively high content of serine proteinases, the venom of C. basiliscus did not exert in vitro coagulant or in vivo defibrinogenating activities. The ability of antivenoms raised against the venoms of C. basiliscus and C. simus (from Costa Rica) to neutralize homologous and heterologous venoms revealed antigenic similarities between toxins of both venoms. Preclinical evaluation of an antivenom produced by using the venom of C. basiliscus as immunogen demonstrated that it is able to neutralize not only the most relevant toxic activities of C. basiliscus venom, but also those exerted by Costa Rican C. simus venom, including coagulant and defibrinogenating activities. The Central American rattlesnake (Crotalus simus) is widely distributed from Mexico to west central Costa Rica, and induces an important number of envenomations in this region. On the other hand, the immunogenic mixture used by Laboratorios de Biológicos y Reactivos de Mexico S.A. (Birmex) to produce the snake antivenom more frequently used in Mexico does not include the venom of C. simus. This immunogenic mixture is composed by the venoms of the Fer-de-lance (Bothrops asper) and the Mexican west-coast rattlesnake (Crotalus basiliscus). We studied the protein composition, toxicological profile and immunogenic properties of the venom of C. basiliscus, and evaluated the ability of the Birmex antivenom to neutralize the venom of C. basiliscus and whether it cross-neutralizes the venom of C. simus from Costa Rica. Using proteomics analysis, in combination with in vitro and mouse tests, we determined that the venom of C. basiliscus is mainly composed by SVMPs, which confer proteolytic and hemorrhagic activities to the venom. Other major components of the venom of C. basiliscus are PLA2s, which are responsible for the myotoxic activity and are the main contributors to the lethal activity. Non-clotting SVSPs correspond to 11% of the venom. Minor components include SVMP-ITs, BPPs, CRISPs and LAAOs, which have not been associated with toxicity. The antibodies induced in horses by the venom of C. basiliscus are able to neutralize not only the most relevant toxic activities of the homologous venom, but also those exerted by Costa Rican C. simus venom, including coagulant and defibrinogenating activities. Our preclinical evaluation suggests that Birmex antivenom can be used to treat envenomations by Costa Rican adult C. simus snakebites, despite this venom not being included in the immunizing mixture.
Copyright © 2017 Elsevier B.V. All rights reserved.
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Publication Date: 2017-02-24 PubMed ID: 28238904DOI: 10.1016/j.jprot.2017.02.015Google Scholar: Lookup
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Summary

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The research article focuses on studying the venom of the Mexican west-coast rattlesnake (Crotalus basiliscus) in terms of its protein composition, toxicity and immune characteristics, and its similarity with the venom of the Central American rattlesnake (Crotalus simus). It investigates the capacity of anti-venoms to neutralize toxic activities of both venoms, suggesting the possibility of using Birmex antivenom to treat envenomations from Costa Rican adult C. simus snakebites.

Protein Composition and Toxicological Profile

  • Identifying the compound makeup of the Mexican west-coast rattlesnake venom, the researchers found that it contains 68% SVMPs, 14% PLAs, 11% serine proteinases, 4% SVMP-Is, 2% BPPs, 0.6% CRISPs, and 0.2% LAAOs.
  • SVMPs present are responsible for azocasein hydrolysis and hemorrhagic activity, contributing to the venom’s lethal activity in mice. However, this is concealed by the neurotoxic action of PLAs, which also cause myotoxic activity.
  • The venom did not exhibit in vitro coagulant or in vivo defibrinogenating activities even though it has a high content of serine proteinases.

Immunogenic Properties and Antivenom Testing

  • Antivenoms created against both C. basiliscus and C. simus venoms were able to neutralize both homologous and heterologous venoms due to antigenic similarities between the toxins in both species’ venom.
  • A preclinical evaluation of an antivenom developed using the C. basiliscus venom as an immunogen displayed its ability to counteract the most toxic activities from both the C.basiliscus and Costa Rican C. simus venom, inclusive of coagulant and defibrinogenating activities.

Comparison Between Venoms and Potential Use of Birmex Antivenom

  • The venom of the Central American rattlesnake C. simus, ranging from Mexico to west central Costa Rica, causes a significant number of envenomations in this region.
  • However, the venom of C. simus is not part of the immunogenic mixture used by the Laboratorios de Biológicos y Reactivos de Mexico S.A. (Birmex) to produce the most frequently used snake antivenom in Mexico.
  • The research concluded that Birmex antivenom, despite not having C. simus venom included in its mixture, has the potential to treat envenomations by Costa Rica’s adult C. simus snakebites.

Cite This Article

APA
Segura Á, Herrera M, Reta Mares F, Jaime C, Sánchez A, Vargas M, Villalta M, Gómez A, Gutiérrez JM, León G. (2017). Proteomic, toxicological and immunogenic characterization of Mexican west-coast rattlesnake (Crotalus basiliscus) venom and its immunological relatedness with the venom of Central American rattlesnake (Crotalus simus). J Proteomics, 158, 62-72. https://doi.org/10.1016/j.jprot.2017.02.015

Publication

Journal of proteomics
ISSN: 1876-7737
NlmUniqueID: 101475056
Country: Netherlands
Language: English
Volume: 158
Pages: 62-72

Researcher Affiliations

Segura, Álvaro
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Herrera, María
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica; Sección de Química Analítica, Escuela de Química, Universidad de Costa Rica, San José, Costa Rica.
Reta Mares, Francisco
  • Laboratorios de Biológicos y Reactivos de México S. A. de C. V., Ciudad de México, Mexico.
Jaime, Claudia
  • Laboratorios de Biológicos y Reactivos de México S. A. de C. V., Ciudad de México, Mexico.
Sánchez, Andrés
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Vargas, Mariángela
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Villalta, Mauren
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Gómez, Aarón
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
Gutiérrez, José María
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica.
León, Guillermo
  • Instituto Clodomiro Picado, Facultad de Microbiología, Universidad de Costa Rica, San José, Costa Rica. Electronic address: guillermo.leon@ucr.ac.cr.

MeSH Terms

  • Animals
  • Crotalus / immunology
  • Crotalus / metabolism
  • Crotoxin / immunology
  • Crotoxin / metabolism
  • Mexico
  • Proteomics
  • Species Specificity

Citations

This article has been cited 21 times.
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