Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D
Loxoscelism is the clinical condition triggered after the bite of spiders of the genus <i>Loxosceles</i>. The main species involved in accidents in South America are <i>L. intermedia</i>, <i>L. laeta,</i> and <i>L. gaucho</i>. The only specific treatme...
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| Format: | Article |
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MDPI AG
2022-12-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/11/1/79 |
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| author | Bruno Cesar Antunes Nayanne Louise Costacurta Polli Pedro Henrique de Caires Schluga Thais Pereira da Silva Ana Carolina Martins Wille Rosangela Locatelli-Dittrich Giovana Scuissiatto de Souza Fernando Hitomi Matsubara João Carlos Minozzo Andrea Senff-Ribeiro Luiza Helena Gremski Silvio Sanches Veiga |
| author_facet | Bruno Cesar Antunes Nayanne Louise Costacurta Polli Pedro Henrique de Caires Schluga Thais Pereira da Silva Ana Carolina Martins Wille Rosangela Locatelli-Dittrich Giovana Scuissiatto de Souza Fernando Hitomi Matsubara João Carlos Minozzo Andrea Senff-Ribeiro Luiza Helena Gremski Silvio Sanches Veiga |
| author_sort | Bruno Cesar Antunes |
| collection | DOAJ |
| description | Loxoscelism is the clinical condition triggered after the bite of spiders of the genus <i>Loxosceles</i>. The main species involved in accidents in South America are <i>L. intermedia</i>, <i>L. laeta,</i> and <i>L. gaucho</i>. The only specific treatment is the anti-<i>Loxosceles</i> serum produced with crude venoms. As phospholipases D (PLDs) trigger most of the effects observed in accidents, we developed and evaluated second-generation sera using mutated PLDs as antigens. Three isoforms of PLDs with site-directed mutations without biological activities were used for rabbit immunizations: D32A-E34A (<i>L. gaucho</i>), W230A (<i>L. intermedia</i>)<i>,</i> and H12A-H47A (<i>L. laeta</i>). Sera were produced using crude venoms of three species of <i>Loxosceles</i> enriched with mutated recombinant PLDs (MIX) or using only mutated PLDs (REC). Immunizations stimulated the immune system from the second immunization with higher antibody production in the REC group. In vivo neutralization assays demonstrated that both sera reduced edema and dermonecrosis caused by <i>Loxosceles intermedia</i> crude venom. Follow-up of animals during the immunization protocols and in the neutralization assays demonstrated that the mutated proteins and the sera are safe. Results demonstrate the potential of using mutated recombinant PLDs in total or partial replacement of <i>Loxosceles</i> venoms in animal immunizations to produce anti-<i>Loxosceles</i> sera for treatments of Loxoscelism. |
| first_indexed | 2024-03-09T13:29:52Z |
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| id | doaj.art-1536bf20ce2c4f13967d084a6ae98602 |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-09T13:29:52Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
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| series | Biomedicines |
| spelling | doaj.art-1536bf20ce2c4f13967d084a6ae986022023-11-30T21:19:08ZengMDPI AGBiomedicines2227-90592022-12-011117910.3390/biomedicines11010079Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-DBruno Cesar Antunes0Nayanne Louise Costacurta Polli1Pedro Henrique de Caires Schluga2Thais Pereira da Silva3Ana Carolina Martins Wille4Rosangela Locatelli-Dittrich5Giovana Scuissiatto de Souza6Fernando Hitomi Matsubara7João Carlos Minozzo8Andrea Senff-Ribeiro9Luiza Helena Gremski10Silvio Sanches Veiga11Department of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilDepartment of Structural, Molecular Biology and Genetics, State University of Ponta Grossa (UEPG), Ponta Grossa 84030-900, BrazilVeterinary Hospital, Federal University of Paraná (UFPR), Curitiba 80035-050, BrazilVeterinary Hospital, Federal University of Paraná (UFPR), Curitiba 80035-050, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilProduction and Research Center of Immunobiological Products (CPPI), State Department of Health, Piraquara 83302-200, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilDepartment of Cell Biology, Federal University of Paraná (UFPR), Curitiba 81530-900, BrazilLoxoscelism is the clinical condition triggered after the bite of spiders of the genus <i>Loxosceles</i>. The main species involved in accidents in South America are <i>L. intermedia</i>, <i>L. laeta,</i> and <i>L. gaucho</i>. The only specific treatment is the anti-<i>Loxosceles</i> serum produced with crude venoms. As phospholipases D (PLDs) trigger most of the effects observed in accidents, we developed and evaluated second-generation sera using mutated PLDs as antigens. Three isoforms of PLDs with site-directed mutations without biological activities were used for rabbit immunizations: D32A-E34A (<i>L. gaucho</i>), W230A (<i>L. intermedia</i>)<i>,</i> and H12A-H47A (<i>L. laeta</i>). Sera were produced using crude venoms of three species of <i>Loxosceles</i> enriched with mutated recombinant PLDs (MIX) or using only mutated PLDs (REC). Immunizations stimulated the immune system from the second immunization with higher antibody production in the REC group. In vivo neutralization assays demonstrated that both sera reduced edema and dermonecrosis caused by <i>Loxosceles intermedia</i> crude venom. Follow-up of animals during the immunization protocols and in the neutralization assays demonstrated that the mutated proteins and the sera are safe. Results demonstrate the potential of using mutated recombinant PLDs in total or partial replacement of <i>Loxosceles</i> venoms in animal immunizations to produce anti-<i>Loxosceles</i> sera for treatments of Loxoscelism.https://www.mdpi.com/2227-9059/11/1/79brown spidervenomloxoscelismserum therapyphospholipases D |
| spellingShingle | Bruno Cesar Antunes Nayanne Louise Costacurta Polli Pedro Henrique de Caires Schluga Thais Pereira da Silva Ana Carolina Martins Wille Rosangela Locatelli-Dittrich Giovana Scuissiatto de Souza Fernando Hitomi Matsubara João Carlos Minozzo Andrea Senff-Ribeiro Luiza Helena Gremski Silvio Sanches Veiga Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D Biomedicines brown spider venom loxoscelism serum therapy phospholipases D |
| title | Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D |
| title_full | Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D |
| title_fullStr | Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D |
| title_full_unstemmed | Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D |
| title_short | Production and Functional Evaluation of Anti-<i>Loxosceles</i> Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D |
| title_sort | production and functional evaluation of anti i loxosceles i sera raised by immunizations of rabbits with mutated recombinant phospholipases d |
| topic | brown spider venom loxoscelism serum therapy phospholipases D |
| url | https://www.mdpi.com/2227-9059/11/1/79 |
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