Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs

Autores
Garcia Denegri, María Emilia; Maruñak, Silvana L.; Todaro, Juan S.; Ponce Soto, Luis A.; Acosta, Ofelia Cristina; Leiva, Laura C.
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Basic phospholipases A2 (PLA2) are toxic and induce a wide spectrum of pharmacological effects, although the acidic enzyme types are not lethal or cause low lethality. Therefore, it is challenging to elucidate the mechanism of action of acidic phospholipases. This study used the acidic non-toxic Ba SpII RP4 PLA2 from Bothrops alternatus as an antigen to develop anti-PLA2 IgG antibodies in rabbits and used in vivo assays to examine the changes in crude venom when pre-incubated with these antibodies. Using Ouchterlony and western blot analyses on B. alternatus venom, we examined the specificity and sensitivity of phospholipase A2 recognition by the specific antibodies (anti-PLA2 IgG). Neutralisation assays using a non-toxic PLA2 antigen revealed unexpected results. The (indirect) haemolytic activity of whole venom was completely inhibited, and all catalytically active phospholipases A2 were blocked. Myotoxicity and lethality were reduced when the crude venom was pre-incubated with anti-PLA2 immunoglobulins. CK levels in the skeletal muscle were significantly reduced at 6 h, and the muscular damage was more significant at this time-point compared to 3 and 12 h. When four times the LD50 was used (224 µg), half the animals treated with the venom-anti PLA2 IgG mixture survived after 48 h. All assays performed with the specific antibodies revealed that Ba SpII RP4 PLA2 had a synergistic effect on whole-venom toxicity. IgG antibodies against the venom of the Argentinean species B. alternatus represent a valuable tool for elucidation of the roles of acidic PLA2 that appear to have purely digestive roles and for further studies on immunotherapy and snake envenoming in affected areas in Argentina and Brazil.
Fil: Garcia Denegri, María Emilia. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias. Departamento de Clinica. Catedra de Farmacologia; Argentina. Universidad Nacional del Nordeste. Facultad de Cs.exactas Naturales y Agrimensura. Departamento de Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Maruñak, Silvana L.. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; Argentina
Fil: Todaro, Juan S.. Universidad Nacional del Nordeste. Facultad de Medicina; Argentina
Fil: Ponce Soto, Luis A.. Universidade Estadual Do Campinas. Instituto de Biologia; Brasil
Fil: Acosta, Ofelia Cristina. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Leiva, Laura C.. Universidad Nacional del Nordeste. Facultad de Cs.exactas Naturales y Agrimensura. Departamento de Bioquimica. Laboratorio de Investigacion En Proteinas; Argentina
Materia
Envenomation
Anti-Pla2 Immunoglobulins
Bothrops Alternatus
Non-Myotoxic Pla2
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/10576
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/10576
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGsGarcia Denegri, María EmiliaMaruñak, Silvana L.Todaro, Juan S.Ponce Soto, Luis A.Acosta, Ofelia CristinaLeiva, Laura C.EnvenomationAnti-Pla2 ImmunoglobulinsBothrops AlternatusNon-Myotoxic Pla2https://purl.org/becyt/ford/4.3https://purl.org/becyt/ford/4Basic phospholipases A2 (PLA2) are toxic and induce a wide spectrum of pharmacological effects, although the acidic enzyme types are not lethal or cause low lethality. Therefore, it is challenging to elucidate the mechanism of action of acidic phospholipases. This study used the acidic non-toxic Ba SpII RP4 PLA2 from Bothrops alternatus as an antigen to develop anti-PLA2 IgG antibodies in rabbits and used in vivo assays to examine the changes in crude venom when pre-incubated with these antibodies. Using Ouchterlony and western blot analyses on B. alternatus venom, we examined the specificity and sensitivity of phospholipase A2 recognition by the specific antibodies (anti-PLA2 IgG). Neutralisation assays using a non-toxic PLA2 antigen revealed unexpected results. The (indirect) haemolytic activity of whole venom was completely inhibited, and all catalytically active phospholipases A2 were blocked. Myotoxicity and lethality were reduced when the crude venom was pre-incubated with anti-PLA2 immunoglobulins. CK levels in the skeletal muscle were significantly reduced at 6 h, and the muscular damage was more significant at this time-point compared to 3 and 12 h. When four times the LD50 was used (224 µg), half the animals treated with the venom-anti PLA2 IgG mixture survived after 48 h. All assays performed with the specific antibodies revealed that Ba SpII RP4 PLA2 had a synergistic effect on whole-venom toxicity. IgG antibodies against the venom of the Argentinean species B. alternatus represent a valuable tool for elucidation of the roles of acidic PLA2 that appear to have purely digestive roles and for further studies on immunotherapy and snake envenoming in affected areas in Argentina and Brazil.Fil: Garcia Denegri, María Emilia. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias. Departamento de Clinica. Catedra de Farmacologia; Argentina. Universidad Nacional del Nordeste. Facultad de Cs.exactas Naturales y Agrimensura. Departamento de Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Maruñak, Silvana L.. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; ArgentinaFil: Todaro, Juan S.. Universidad Nacional del Nordeste. Facultad de Medicina; ArgentinaFil: Ponce Soto, Luis A.. Universidade Estadual Do Campinas. Instituto de Biologia; BrasilFil: Acosta, Ofelia Cristina. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Leiva, Laura C.. Universidad Nacional del Nordeste. Facultad de Cs.exactas Naturales y Agrimensura. Departamento de Bioquimica. Laboratorio de Investigacion En Proteinas; ArgentinaElsevier2014-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/10576Garcia Denegri, María Emilia; Maruñak, Silvana L.; Todaro, Juan S.; Ponce Soto, Luis A.; Acosta, Ofelia Cristina; et al.; Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs; Elsevier; Toxicon; 86; 5-2014; 89-950041-0101enginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0041010114001500info:eu-repo/semantics/altIdentifier/doi/10.1016/j.toxicon.2014.05.016info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:13:29Zoai:ri.conicet.gov.ar:11336/10576instacron: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:34982025-10-15 15:13:29.36CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
title Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
spellingShingle Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
Garcia Denegri, María Emilia
Envenomation
Anti-Pla2 Immunoglobulins
Bothrops Alternatus
Non-Myotoxic Pla2
title_short Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
title_full Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
title_fullStr Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
title_full_unstemmed Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
title_sort Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs
dc.creator.none.fl_str_mv Garcia Denegri, María Emilia
Maruñak, Silvana L.
Todaro, Juan S.
Ponce Soto, Luis A.
Acosta, Ofelia Cristina
Leiva, Laura C.
author Garcia Denegri, María Emilia
author_facet Garcia Denegri, María Emilia
Maruñak, Silvana L.
Todaro, Juan S.
Ponce Soto, Luis A.
Acosta, Ofelia Cristina
Leiva, Laura C.
author_role author
author2 Maruñak, Silvana L.
Todaro, Juan S.
Ponce Soto, Luis A.
Acosta, Ofelia Cristina
Leiva, Laura C.
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Envenomation
Anti-Pla2 Immunoglobulins
Bothrops Alternatus
Non-Myotoxic Pla2
topic Envenomation
Anti-Pla2 Immunoglobulins
Bothrops Alternatus
Non-Myotoxic Pla2
purl_subject.fl_str_mv https://purl.org/becyt/ford/4.3
https://purl.org/becyt/ford/4
dc.description.none.fl_txt_mv Basic phospholipases A2 (PLA2) are toxic and induce a wide spectrum of pharmacological effects, although the acidic enzyme types are not lethal or cause low lethality. Therefore, it is challenging to elucidate the mechanism of action of acidic phospholipases. This study used the acidic non-toxic Ba SpII RP4 PLA2 from Bothrops alternatus as an antigen to develop anti-PLA2 IgG antibodies in rabbits and used in vivo assays to examine the changes in crude venom when pre-incubated with these antibodies. Using Ouchterlony and western blot analyses on B. alternatus venom, we examined the specificity and sensitivity of phospholipase A2 recognition by the specific antibodies (anti-PLA2 IgG). Neutralisation assays using a non-toxic PLA2 antigen revealed unexpected results. The (indirect) haemolytic activity of whole venom was completely inhibited, and all catalytically active phospholipases A2 were blocked. Myotoxicity and lethality were reduced when the crude venom was pre-incubated with anti-PLA2 immunoglobulins. CK levels in the skeletal muscle were significantly reduced at 6 h, and the muscular damage was more significant at this time-point compared to 3 and 12 h. When four times the LD50 was used (224 µg), half the animals treated with the venom-anti PLA2 IgG mixture survived after 48 h. All assays performed with the specific antibodies revealed that Ba SpII RP4 PLA2 had a synergistic effect on whole-venom toxicity. IgG antibodies against the venom of the Argentinean species B. alternatus represent a valuable tool for elucidation of the roles of acidic PLA2 that appear to have purely digestive roles and for further studies on immunotherapy and snake envenoming in affected areas in Argentina and Brazil.
Fil: Garcia Denegri, María Emilia. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias. Departamento de Clinica. Catedra de Farmacologia; Argentina. Universidad Nacional del Nordeste. Facultad de Cs.exactas Naturales y Agrimensura. Departamento de Agrimensura; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Maruñak, Silvana L.. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; Argentina
Fil: Todaro, Juan S.. Universidad Nacional del Nordeste. Facultad de Medicina; Argentina
Fil: Ponce Soto, Luis A.. Universidade Estadual Do Campinas. Instituto de Biologia; Brasil
Fil: Acosta, Ofelia Cristina. Universidad Nacional del Nordeste. Facultad de Cs.veterinarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Leiva, Laura C.. Universidad Nacional del Nordeste. Facultad de Cs.exactas Naturales y Agrimensura. Departamento de Bioquimica. Laboratorio de Investigacion En Proteinas; Argentina
description Basic phospholipases A2 (PLA2) are toxic and induce a wide spectrum of pharmacological effects, although the acidic enzyme types are not lethal or cause low lethality. Therefore, it is challenging to elucidate the mechanism of action of acidic phospholipases. This study used the acidic non-toxic Ba SpII RP4 PLA2 from Bothrops alternatus as an antigen to develop anti-PLA2 IgG antibodies in rabbits and used in vivo assays to examine the changes in crude venom when pre-incubated with these antibodies. Using Ouchterlony and western blot analyses on B. alternatus venom, we examined the specificity and sensitivity of phospholipase A2 recognition by the specific antibodies (anti-PLA2 IgG). Neutralisation assays using a non-toxic PLA2 antigen revealed unexpected results. The (indirect) haemolytic activity of whole venom was completely inhibited, and all catalytically active phospholipases A2 were blocked. Myotoxicity and lethality were reduced when the crude venom was pre-incubated with anti-PLA2 immunoglobulins. CK levels in the skeletal muscle were significantly reduced at 6 h, and the muscular damage was more significant at this time-point compared to 3 and 12 h. When four times the LD50 was used (224 µg), half the animals treated with the venom-anti PLA2 IgG mixture survived after 48 h. All assays performed with the specific antibodies revealed that Ba SpII RP4 PLA2 had a synergistic effect on whole-venom toxicity. IgG antibodies against the venom of the Argentinean species B. alternatus represent a valuable tool for elucidation of the roles of acidic PLA2 that appear to have purely digestive roles and for further studies on immunotherapy and snake envenoming in affected areas in Argentina and Brazil.
publishDate 2014
dc.date.none.fl_str_mv 2014-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/10576
Garcia Denegri, María Emilia; Maruñak, Silvana L.; Todaro, Juan S.; Ponce Soto, Luis A.; Acosta, Ofelia Cristina; et al.; Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs; Elsevier; Toxicon; 86; 5-2014; 89-95
0041-0101
url http://hdl.handle.net/11336/10576
identifier_str_mv Garcia Denegri, María Emilia; Maruñak, Silvana L.; Todaro, Juan S.; Ponce Soto, Luis A.; Acosta, Ofelia Cristina; et al.; Neutralisation of the pharmacological activities of Bothrops alternatus venom by anti-PLA2 IgGs; Elsevier; Toxicon; 86; 5-2014; 89-95
0041-0101
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S0041010114001500
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.toxicon.2014.05.016
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
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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score 13.22299

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