| Full text | |
| Author(s): |
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]
Total Authors: 9
|
| Affiliation: | [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
Total Affiliations: 2
|
| Document type: | Journal article |
| Source: | Toxicon; v. 169, p. 59-67, NOV 2019. |
| Web of Science Citations: | 0 |
| Abstract | |
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) | |
| FAPESP's process: | 15/13124-5 - Purification and characterization of peptides present in the venom of African snakes: searching for peptidase inhibitor of medical importance |
| Grantee: | Roberto Tadashi Kodama |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 13/07467-1 - CeTICS - Center of Toxins, Immune-Response and Cell Signaling |
| Grantee: | Hugo Aguirre Armelin |
| Support Opportunities: | Research Grants - Research, Innovation and Dissemination Centers - RIDC |
| FAPESP's process: | 13/15344-7 - Efficacy of the bothropic antivenom from Butantan Institute: obtention, characterization and neutralization of serinepeptidases from the venom of Bothrops jararaca |
| Grantee: | Alexandre Kazuo Kuniyoshi |
| Support Opportunities: | Scholarships in Brazil - Doctorate |
| FAPESP's process: | 12/06677-0 - Study of peptidase activity of the B. jararaca venom and potential neutralizing serum produced at Instituto Butantan upon this activity: new aspects of Bothrops poisoning |
| Grantee: | Fernanda Calheta Vieira Portaro |
| Support Opportunities: | Regular Research Grants |
| FAPESP's process: | 15/15364-3 - Toxic potential analysis of proteases and peptides present in scorpion Tityus serrulatus venom and the blockage capacity of commercial antivenoms: Enhancing the knowledge of venom and its mechanism of action. |
| Grantee: | Fernanda Calheta Vieira Portaro |
| Support Opportunities: | Regular Research Grants |