Bothrops alternatus venom pretrated with chelate agent for antiserum production

Autores
López, Gisela Lumila; Van de Velde, Andrea Carolina; Leiva, Laura Cristina Ana; Fusco, Luciano Sebastian
Año de publicación
2018
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Snakebite envenoming is a major public health problem in tropical countries. The only specific therapies currently available for the treatment are antivenoms, which consist of polyclonal immunoglobulins purified from sera/plasma of horses or sheep immunized with doses sublethal of snake venom(s). Bothrops alternatus (BaV) venom induces a prominent tissue local damage, especially, hemorrhage, muscle damage and inflammation. These complex pathological phenomena are due to the concomitant action of metalloproteinases (SVMPs) in venoms and others edema-inducing components. In order to obtain a high titer of antibodies and reduced local damage in the animal, an alternative immunization protocol was proposed where the SVMPs action was blocked by using a chelate agent. For this proposal, the B. alternatus venom (10 mg/mL) was blocked by EDTA-Na2 (10 mM, BaV/ EDTA-Na2) and used as antigen. Previously to the inoculation, the excess of chelate was removed by passing the mixture on Sephadex G-25 column (venom without inhibitor was subjected to the same process) and the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 Balb/c mice were immunized subcutaneously on days 0, 15 and 30 with BaV(15-30-45μg) or BaV/ EDTA-Na2 (45-90-135 μg) emulsified with Freund?s Adyuvant (complete first and incomplete-booster). Blood samples were collected by the animals tail tip on days 14, 29 and 41 of protocol immunization and it was destined to ELISA´s test. The results showed that the immunized animal with BaV/ EDTA-Na2had a higher titer (5.1x104) than those treated to BaV (1.3x104). Macroscopic analysis at the inoculation site of mice injected with Freund?s adjuvant showed local damage (with non-infectious abscesses) and hypertrophy of inguinal lymph nodes. Our results show that BaV/ EDTA-Na2 formulation, where the SVMPs are blocked, produced a higher humoral response compared with the produced by BaV. These preliminary results demonstrated the potential use of blocking the toxins with chelate to produce antivenom with less damage in the animals.
Fil: López, Gisela Lumila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
Fil: Van de Velde, Andrea Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Fusco, Luciano Sebastian. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
XX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of Biosciences
Buenos Aires
Argentina
Sociedad Argentina de Biología
Sociedad Uruguaya de Biociencias
Materia
ANTIVENOM
EDTA
METALOPROTEASE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/181336
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Bothrops alternatus venom pretrated with chelate agent for antiserum productionLópez, Gisela LumilaVan de Velde, Andrea CarolinaLeiva, Laura Cristina AnaFusco, Luciano SebastianANTIVENOMEDTAMETALOPROTEASEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Snakebite envenoming is a major public health problem in tropical countries. The only specific therapies currently available for the treatment are antivenoms, which consist of polyclonal immunoglobulins purified from sera/plasma of horses or sheep immunized with doses sublethal of snake venom(s). Bothrops alternatus (BaV) venom induces a prominent tissue local damage, especially, hemorrhage, muscle damage and inflammation. These complex pathological phenomena are due to the concomitant action of metalloproteinases (SVMPs) in venoms and others edema-inducing components. In order to obtain a high titer of antibodies and reduced local damage in the animal, an alternative immunization protocol was proposed where the SVMPs action was blocked by using a chelate agent. For this proposal, the B. alternatus venom (10 mg/mL) was blocked by EDTA-Na2 (10 mM, BaV/ EDTA-Na2) and used as antigen. Previously to the inoculation, the excess of chelate was removed by passing the mixture on Sephadex G-25 column (venom without inhibitor was subjected to the same process) and the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 Balb/c mice were immunized subcutaneously on days 0, 15 and 30 with BaV(15-30-45μg) or BaV/ EDTA-Na2 (45-90-135 μg) emulsified with Freund?s Adyuvant (complete first and incomplete-booster). Blood samples were collected by the animals tail tip on days 14, 29 and 41 of protocol immunization and it was destined to ELISA´s test. The results showed that the immunized animal with BaV/ EDTA-Na2had a higher titer (5.1x104) than those treated to BaV (1.3x104). Macroscopic analysis at the inoculation site of mice injected with Freund?s adjuvant showed local damage (with non-infectious abscesses) and hypertrophy of inguinal lymph nodes. Our results show that BaV/ EDTA-Na2 formulation, where the SVMPs are blocked, produced a higher humoral response compared with the produced by BaV. These preliminary results demonstrated the potential use of blocking the toxins with chelate to produce antivenom with less damage in the animals.Fil: López, Gisela Lumila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; ArgentinaFil: Van de Velde, Andrea Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaFil: Fusco, Luciano Sebastian. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; ArgentinaXX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of BiosciencesBuenos AiresArgentinaSociedad Argentina de BiologíaSociedad Uruguaya de BiocienciasTech Science Press2018info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/181336Bothrops alternatus venom pretrated with chelate agent for antiserum production; XX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of Biosciences; Buenos Aires; Argentina; 2018; 1-20327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/biocell/v43nSuppl.3/33866Internacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:09:31Zoai:ri.conicet.gov.ar:11336/181336instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:09:31.331CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Bothrops alternatus venom pretrated with chelate agent for antiserum production
title Bothrops alternatus venom pretrated with chelate agent for antiserum production
spellingShingle Bothrops alternatus venom pretrated with chelate agent for antiserum production
López, Gisela Lumila
ANTIVENOM
EDTA
METALOPROTEASE
title_short Bothrops alternatus venom pretrated with chelate agent for antiserum production
title_full Bothrops alternatus venom pretrated with chelate agent for antiserum production
title_fullStr Bothrops alternatus venom pretrated with chelate agent for antiserum production
title_full_unstemmed Bothrops alternatus venom pretrated with chelate agent for antiserum production
title_sort Bothrops alternatus venom pretrated with chelate agent for antiserum production
dc.creator.none.fl_str_mv López, Gisela Lumila
Van de Velde, Andrea Carolina
Leiva, Laura Cristina Ana
Fusco, Luciano Sebastian
author López, Gisela Lumila
author_facet López, Gisela Lumila
Van de Velde, Andrea Carolina
Leiva, Laura Cristina Ana
Fusco, Luciano Sebastian
author_role author
author2 Van de Velde, Andrea Carolina
Leiva, Laura Cristina Ana
Fusco, Luciano Sebastian
author2_role author
author
author
dc.subject.none.fl_str_mv ANTIVENOM
EDTA
METALOPROTEASE
topic ANTIVENOM
EDTA
METALOPROTEASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Snakebite envenoming is a major public health problem in tropical countries. The only specific therapies currently available for the treatment are antivenoms, which consist of polyclonal immunoglobulins purified from sera/plasma of horses or sheep immunized with doses sublethal of snake venom(s). Bothrops alternatus (BaV) venom induces a prominent tissue local damage, especially, hemorrhage, muscle damage and inflammation. These complex pathological phenomena are due to the concomitant action of metalloproteinases (SVMPs) in venoms and others edema-inducing components. In order to obtain a high titer of antibodies and reduced local damage in the animal, an alternative immunization protocol was proposed where the SVMPs action was blocked by using a chelate agent. For this proposal, the B. alternatus venom (10 mg/mL) was blocked by EDTA-Na2 (10 mM, BaV/ EDTA-Na2) and used as antigen. Previously to the inoculation, the excess of chelate was removed by passing the mixture on Sephadex G-25 column (venom without inhibitor was subjected to the same process) and the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 Balb/c mice were immunized subcutaneously on days 0, 15 and 30 with BaV(15-30-45μg) or BaV/ EDTA-Na2 (45-90-135 μg) emulsified with Freund?s Adyuvant (complete first and incomplete-booster). Blood samples were collected by the animals tail tip on days 14, 29 and 41 of protocol immunization and it was destined to ELISA´s test. The results showed that the immunized animal with BaV/ EDTA-Na2had a higher titer (5.1x104) than those treated to BaV (1.3x104). Macroscopic analysis at the inoculation site of mice injected with Freund?s adjuvant showed local damage (with non-infectious abscesses) and hypertrophy of inguinal lymph nodes. Our results show that BaV/ EDTA-Na2 formulation, where the SVMPs are blocked, produced a higher humoral response compared with the produced by BaV. These preliminary results demonstrated the potential use of blocking the toxins with chelate to produce antivenom with less damage in the animals.
Fil: López, Gisela Lumila. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina
Fil: Van de Velde, Andrea Carolina. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Leiva, Laura Cristina Ana. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
Fil: Fusco, Luciano Sebastian. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Departamento de Bioquímica. Laboratorio de Investigación en Proteínas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Química Básica y Aplicada del Nordeste Argentino. Universidad Nacional del Nordeste. Facultad de Ciencias Exactas Naturales y Agrimensura. Instituto de Química Básica y Aplicada del Nordeste Argentino; Argentina
XX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of Biosciences
Buenos Aires
Argentina
Sociedad Argentina de Biología
Sociedad Uruguaya de Biociencias
description Snakebite envenoming is a major public health problem in tropical countries. The only specific therapies currently available for the treatment are antivenoms, which consist of polyclonal immunoglobulins purified from sera/plasma of horses or sheep immunized with doses sublethal of snake venom(s). Bothrops alternatus (BaV) venom induces a prominent tissue local damage, especially, hemorrhage, muscle damage and inflammation. These complex pathological phenomena are due to the concomitant action of metalloproteinases (SVMPs) in venoms and others edema-inducing components. In order to obtain a high titer of antibodies and reduced local damage in the animal, an alternative immunization protocol was proposed where the SVMPs action was blocked by using a chelate agent. For this proposal, the B. alternatus venom (10 mg/mL) was blocked by EDTA-Na2 (10 mM, BaV/ EDTA-Na2) and used as antigen. Previously to the inoculation, the excess of chelate was removed by passing the mixture on Sephadex G-25 column (venom without inhibitor was subjected to the same process) and the effective neutralization of SVMPs using azocasein as substrate was determined. Group of 5 Balb/c mice were immunized subcutaneously on days 0, 15 and 30 with BaV(15-30-45μg) or BaV/ EDTA-Na2 (45-90-135 μg) emulsified with Freund?s Adyuvant (complete first and incomplete-booster). Blood samples were collected by the animals tail tip on days 14, 29 and 41 of protocol immunization and it was destined to ELISA´s test. The results showed that the immunized animal with BaV/ EDTA-Na2had a higher titer (5.1x104) than those treated to BaV (1.3x104). Macroscopic analysis at the inoculation site of mice injected with Freund?s adjuvant showed local damage (with non-infectious abscesses) and hypertrophy of inguinal lymph nodes. Our results show that BaV/ EDTA-Na2 formulation, where the SVMPs are blocked, produced a higher humoral response compared with the produced by BaV. These preliminary results demonstrated the potential use of blocking the toxins with chelate to produce antivenom with less damage in the animals.
publishDate 2018
dc.date.none.fl_str_mv 2018
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Journal
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/181336
Bothrops alternatus venom pretrated with chelate agent for antiserum production; XX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of Biosciences; Buenos Aires; Argentina; 2018; 1-2
0327-9545
1667-5746
CONICET Digital
CONICET
url http://hdl.handle.net/11336/181336
identifier_str_mv Bothrops alternatus venom pretrated with chelate agent for antiserum production; XX Annual Meeting of the Argentinean Biology Society and VII Meeting of the Uruguayan Society of Biosciences; Buenos Aires; Argentina; 2018; 1-2
0327-9545
1667-5746
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.techscience.com/biocell/v43nSuppl.3/33866
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
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application/pdf
dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Tech Science Press
publisher.none.fl_str_mv Tech Science Press
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
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instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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