Antivenomics as a tool to improve the neutralizing capacity of the crotalic antivenom: a study with crotamine

 

Ricardo Teixeira-Araújo^1 2, Patrícia Castanheira¹, Leonora Brazil-Más², Francisco Pontes^1 2, Moema Leitão de Araújo³, Maria Lucia Machado Alves³, Russolina Benedeta Zingali¹, Carlos Correa-Netto^1 2

 

1 Laboratório de Hemostase e Venenos, Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ, Brasil

2 Departamento de Antígenos e Cultivo Celular, Instituto Vital Brazil, Niterói, RJ, Brasil

 

ABSTRACT

Background

Snakebite treatment requires administration of an appropriate antivenom that should contain antibodies capable of neutralizing the venom. To achieve this goal, antivenom production must start from a suitable immunization protocol and proper venom mixtures. In Brazil, antivenom against South American rattlesnake (Crotalus durissus terrificus) bites is produced by public institutions based on the guidelines defined by the regulatory agency of the Brazilian Ministry of Health, ANVISA. However, each institution uses its own mixture of rattlesnake venom antigens. Previous works have shown that crotamine, a toxin found in Crolatus durissus venom, shows marked individual and populational variation. In addition, serum produced from crotamine-negative venoms fails to recognize this molecule.

Methods

In this work, we used an antivenomics approach to assess the cross-reactivity of crotalic antivenom manufactured by IVB towards crotamine-negative venom and a mixture of crotamine-negative/crotamine-positive venoms.

Results

We show that the venom mixture containing 20% crotamine and 57% crotoxin produced a strong immunogenic response in horses. Antivenom raised against this venom mixture reacted with most venom components including crotamine and crotoxin, in contrast to the antivenom raised against crotamine-negative venom.

Conclusions

These results indicate that venomic databases and antivenomics analysis provide a useful approach for choosing the better venom mixture for antibody production and for the subsequent screening of antivenom cross-reactivity with relevant snake venom components.

 

Key words: Antivenom production; Antivenomics; Crotalus durissus; Crotamine; Crotoxin; Geographic venom variation

 

Funding

The present work was supported by the National Council for Scientific and Technological Development (CNPq), Rio de Janeiro Research Foundation (FAPERJ) and by the Edital Toxinologia CAPES no. 063/2010, Process no. 2308.006277/2011-85, AUXPE 1517/2011.

 

Received: December 22, 2016.

Accepted: May 5, 2017.

Revised: May 12, 2017.

 

Correspondence: netto@bioqmed.ufrj.br

 

Authors’ contributions

RTA participated in data collection and experimental design. PC, LBM and FP participated in data collection. MLA and MLMA supervised the collection of Crotalus venom. RBZ controlled the experiments, interpreted results and reviewed the article. CCN performed data collection, experimental design, interpreted results, wrote the article draft, reviewed and edited the article. All authors read and approved the final manuscript.

 

Competing interests

The authors declare that they have no competing interests.

 

Consent for publication

Not applicable.

 

Ethics approval and consent to participate

Not applicable.

 

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