Autor(es)

Kuniyoshi, Alexandre Kazuo ^[1] ; Kodama, Roberto Tadashi ^[1] ; Cajado-Carvalho, Daniela ^[1] ; Iwai, Leo Kei ^[2] ; Kitano, Eduardo ^[2] ; Fernandes da Silva, Cristiane Castilho ^[1] ; Duzzi, Bruno ^[1] ; da Silva, Wilmar Dias ^[1] ; Portaro, Fernanda Calheta ^[1]

Afiliação do(s) autor(es)

^[1] Butantan Inst, Immunochem Lab, Av Prof Vital Brazil 1500, BR-05503900 Sao Paulo, SP - Brazil

^[2] Butantan Inst, Ctr Toxins Immune Response & Cell Signaling CeTIS, Special Lab Appl Toxinol, Sao Paulo, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Toxicon; v. 169, p. 59-67, NOV 2019.
• Citações Web of Science: 0

Resumo

In Brazil, snakes from the Bothrops genus are responsible for thousands of accidents, and their venoms are mainly made up of proteolytic enzymes. Although the antibothropic serum produced by the Butantan Institute is remarkable in saving lives, studies show that some symptoms observed in cases of envenoming are not efficiently neutralized. Moreover, our group has shown that the commercial antivenom does not fully neutralize in vitro some serine proteases present in the Bothrops jararaca venom. Therefore, this study focuses on a new method in the production of specific immunoglobulins capable of neutralizing the activities of these enzymes in vitro. For this, a pool of serine proteases that was not inhibited by the commercial antivenom, made up of four enzymes (KN-BJ2, BjSP, HS112 and BPA) from the B. jararaca venom was obtained through two chromatographic steps (DEAE-HPLC and C8-RP-HPLC). The identities of these proteases were confirmed by SDS-PAGE, followed by tryptic digestion and mass spectrometry analysis. This pool was inoculated into BALB/c and C57BL/6 mice, using SBA-15 as adjuvant, and the produced IgGs were purified by affinity chromatography. The sera were characterized by ELISA, avidity and proteolytic neutralization assays. Both animal models responded to the immunization, producing higher IgGs titers when compared to the commercial antivenom. The experimental serum from BALB/c mice presented a better hydrolysis inhibition of the selective fluorescent substrate for serine proteases (similar to 80%) when compared to C57BL/6 (similar to 25%) and the commercial antivenom ( < 1%) at the dose of 500:1 (weight of antivenom:weight of venom). These results show that a different immunization method using isolated serine proteases improves the toxins neutralizing efficacy and could lead to a better end product to be used as a supplemental medicine to the currently used immunotherapy. (AU)

• Processo FAPESP: 15/13124-5 - Purificação e caracterização de peptídeos presentes nos venenos de serpentes africanas: à procura de inibidores de peptidases de importância médica
• Beneficiário: Roberto Tadashi Kodama
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 13/07467-1 - CeTICS - Centro de Toxinas, Imuno-Resposta e Sinalização Celular
• Beneficiário: Hugo Aguirre Armelin
• Modalidade de apoio: Auxílio à Pesquisa - Centros de Pesquisa, Inovação e Difusão - CEPIDs
• Processo FAPESP: 13/15344-7 - Eficácia do soro antibotrópico produzido no Instituto Butantan: obtenção, caracterização e neutralização de serinopeptidases de interesse do veneno de Bothrops jararaca
• Beneficiário: Alexandre Kazuo Kuniyoshi
• Modalidade de apoio: Bolsas no Brasil - Doutorado
• Processo FAPESP: 12/06677-0 - Estudo da atividade peptidásica dos venenos botrópicos e do potencial neutralizante do soro produzido no Instituto Butantan sobre esta atividade: novos aspectos sobre o envenenamento botrópico
• Beneficiário: Fernanda Calheta Vieira Portaro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/15364-3 - Análise do potencial tóxico de proteases e peptídeos presentes no veneno do escorpião Tityus serrulatus e do poder neutralizante dos antivenenos comerciais: Aprimorando o conhecimento do veneno e seu mecanismo de ação.
• Beneficiário: Fernanda Calheta Vieira Portaro
• Modalidade de apoio: Auxílio à Pesquisa - Regular