Potential low-impact immunogen for the production of anti-bothropic serum: Bothrops alternatus venom treated with Na 2 EDTA.

Autor: Lopez GL; Laboratorio de Investigación en Proteínas (LabInPro), IQUIBA-NEA (CONICET; UNNE), Corrientes, Argentina., Van de Velde A; Laboratorio de Investigación en Proteínas (LabInPro), IQUIBA-NEA (CONICET; UNNE), Corrientes, Argentina., Hernández D; Cátedra de Histología y Embriología, Facultad de Ciencias Veterinarias (FCV), Universidad Nacional del Nordeste (UNNE), Sargento Cabral N° 2139, Corrientes, Argentina., Bustillo S; Grupo de Investigaciones Biológicas y Moleculares (GIBYM), IQUIBA-NEA (CONICET; UNNE), Corrientes, Argentina; Facultad de Ciencias Exactas Naturales y Agrimensura (FaCENA-UNNE), Av. Liberta, 5470, Corrientes, Argentina., Leiva L; Laboratorio de Investigación en Proteínas (LabInPro), IQUIBA-NEA (CONICET; UNNE), Corrientes, Argentina; Facultad de Ciencias Exactas Naturales y Agrimensura (FaCENA-UNNE), Av. Liberta, 5470, Corrientes, Argentina., Fusco LS; Laboratorio de Investigación en Proteínas (LabInPro), IQUIBA-NEA (CONICET; UNNE), Corrientes, Argentina; Facultad de Ciencias Exactas Naturales y Agrimensura (FaCENA-UNNE), Av. Liberta, 5470, Corrientes, Argentina. Electronic address: luciano.fusco@comunidad.unne.edu.ar.
Jazyk: angličtina
Zdroj: Toxicon : official journal of the International Society on Toxinology [Toxicon] 2024 Jan; Vol. 237, pp. 107351. Date of Electronic Publication: 2023 Nov 19.
DOI: 10.1016/j.toxicon.2023.107351
Abstrakt: This study proposes an alternative method using Na 2 EDTA to neutralize B. alternatus venom and using it as an immunogen from the start of inoculation to minimize side effects and enhance antivenom production. To achieve this, 1.8 mg/mL of B. alternatus venom (B.aV) was treated with Na 2 EDTA, and any extra chelate was eliminated by filtering the resulting solution through a Sephadex G-25 column. Two groups of BALB/c mice were immunized subcutaneously on days 1, 15 and 30 with B.aV/Na 2 EDTA (45, 90, 135 μg/mouse) or B.aV (15, 30, 45 μg/mouse), respectively. Both formulations were emulsified with Freund's adjuvant (complete first and incomplete-booster). Blood samples were collected from each mouse on days 14, 29, 41, and 50 post-first immunization, and serum was separated for antibody detection. Animals were then sacrificed and lungs removed for histological analysis (hematoxylin-eosin). Immunoblotting analysis revealed that the sera from mice inoculated with B.aV/Na 2 EDTA (anti-B.aV/Na 2 EDTA) recognized the major venom proteins (20-66 kDa) similarly to the sera from mice inoculated with B.aV (anti-B.aV). The enzyme-linked immunosorbent assay results indicated that the anti-B.aV/Na 2 EDTA had a higher titer (5.76 × 10 4 ) than those the anti-B.aV (1.92 × 10 4 ). Additionally, sera from animals immunized with B.aV/Na 2 EDTA significantly neutralized proteolytic, indirect hemolytic and coagulant activity (p < 0.05). Finally, histological examination of the lungs of mice inoculated with B.aV/Na 2 EDTA showed normal appearance, while animals inoculated with B.aV showed interstitial lung injury (p < 0.05). In conclusion, the B.aV/Na 2 EDTA formulation, free of excess Na 2 EDTA, proved to be a promising candidate as an immunogen for antivenom production.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 Elsevier Ltd. All rights reserved.)
Databáze: MEDLINE
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