Production and Functional Evaluation of Anti- Loxosceles Sera Raised by Immunizations of Rabbits with Mutated Recombinant Phospholipases-D

• Published in: Biomedicines Vol. 11; no. 1; p. 79
• Main Authors: Antunes, Bruno Cesar, Polli, Nayanne Louise Costacurta, Schluga, Pedro Henrique de Caires, Silva, Thais Pereira da, Wille, Ana Carolina Martins, Locatelli-Dittrich, Rosangela, Souza, Giovana Scuissiatto de, Matsubara, Fernando Hitomi, Minozzo, João Carlos, Senff-Ribeiro, Andrea, Gremski, Luiza Helena, Veiga, Silvio Sanches
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 28.12.2022; MDPI
• Subjects: Accidents; Amino acids; Animals; Antibiotics; Antigens; brown spider; Edema; Enzymes; Immune system; Immunization; Isoforms; Loxosceles; loxoscelism; Mutation; phospholipases D; serum therapy; Spiders; Toxins; Venom; Brazil; United States > US; loxoscelism; serum therapy; venom; brown spider; phospholipases D
• ISSN: 2227-9059; 2227-9059
• DOI: 10.3390/biomedicines11010079

Loxoscelism is the clinical condition triggered after the bite of spiders of the genus . The main species involved in accidents in South America are , and . The only specific treatment is the anti- 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 ( ), W230A ( ) and H12A-H47A ( ). Sera were produced using crude venoms of three species of 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 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 venoms in animal immunizations to produce anti- sera for treatments of Loxoscelism.