Can anti-bothropstoxin-I antibodies discriminate between Bothrops jararaca and Bothrops jararacussu venoms?

 

Ricardo Teixeira Araujo^1 2, Carlos Corrêa-Netto^1 2, Leonora Brazil-Más², Caio Raony Farina Silveira³, Irene Fernandes³, Russolina Benedeta Zingali¹

 

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 Instituto Vital Brazil, Niterói, RJ, Brasil.

3 Instituto Butantan, São Paulo, SP, Brasil.

 

ABSTRACT

Background

Snakes of the genus Bothrops, popularly known as pit vipers, are responsible for most cases of snakebite in Brazil. Within this genus, Bothrops jararacussu and B. jararaca deserve special attention due to the severity of their bites and for inhabiting densely populated areas. Regarding the treatment of snakebites by Bothrops jararacussu, questions have been raised about the effectiveness of the specific bothropic antivenom in neutralizing myotoxic effects; however, there are no accurate data for humans. Thus, the development of a differential diagnostic kit for this species would be of great interest because it provides, for healthcare professionals, a tool that would allow us to determine whether the accident was caused by B. jararacussu or other species of the genus. It would also make it possible to evaluate the specificity of the treatment and to provide data for epidemiological studies.

 

Methods

First, we produced a species-specific polyclonal antibody – a potential biomarker of Bothrops jararacussu venom – against bothropstoxin-I (BthTx-I), which is also found in smaller quantities in the venoms of B. jararaca from southern Brazil.

 

Results

Polyclonal antibodies against bothropstoxin-I could be separated into several species-specific immunoglobulins. Then, aiming to develop a system of safe and standardized immunoassay, we produced monoclonal antibodies. Seven hybridomas were obtained. Five of them were specific to the venom of B. jararacussu and two recognized the venom of B. jararaca from the southeastern population. The use of monoclonal antibodies also made it possible to differentiate B. jararacussu from B. jararaca venom obtained from the southern population. Analyzing the reactivity of monoclonal antibodies against other bothropic venoms, we found mAb Bt-3 to be more specific than others for B. jararacussu venom.

 

Conclusions

These results show the potential of BthTx-I for producing monoclonal antibodies that differentiate between B. jararacussu and other Bothrops species venoms.

 

Key words: Bothrops jararacussu; Bothrops jararaca; Bothropstoxin-I; Monoclonal antibodies; Species specificity.

 

Funding

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

Received: September 22, 2016.

Revised: February 24, 2017.

Accepted: March 11, 2017.

 

Correspondence: lzingali@bioqmed.ufrj.br; lzingali@gmail.com

 

Authors’ contributions

RTA wrote the article draft, performed the data collection, analysis and interpretetion. CCN participate in experimental design edited and revised the article. LBM and CRFS participated in data collection and critical revision of the article. IF supervised the experiments on monoclonal antibodies and performed a critical revision of the article. RBZ supervised the experiments, contributed to development of concept and experimental design, writing, reviewing and editing 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

All animal procedures were approved by the Ethics Committee for Animal Research of Butantan Institute (process no. 697/10).