Neutralization of dermonecrotic and lethal activities and differences among 32-35 kDa toxins of medically important Loxosceles spider venoms in Brazil revealed by monoclonal antibodies.
Neutralization of dermonecrotic and lethal activities and differences among the principal toxic proteins (32-35 kDa) of medically important Loxosceles spider venoms in Brazil (Loxosceles gaucho, Loxosceles laeta and Loxosceles intermedia) were studied using monoclonal antibodies (MAbs) produced against the dermonecrotic component (35 kDa) of L. gaucho venom. MAb titers were 512,000 to homologous venom, between 2000 and 64,000 for L. intermedia venom and between 1000 and 64,000 for L. laeta venom. By Western blotting, MAbs could recognize mainly the 35 kDa protein of L. gaucho venom and with less intensity the 35 kDa protein of L. intermedia venom. These MAbs also recognized weakly or did not recognize the 32 kDa component of L. laeta venom. Only MoALg1 showed high affinity for L. gaucho venom and neutralized in vivo 90-97% of the dermonecrotic activity, besides delaying the lethality induced by homologous venom. MoALg1 maintained its capacity to neutralize the dermonecrotic activity, even when administered (i.v.) 6h after envenoming (i.d.). All MAbs obtained failed to neutralize the toxic activities of the heterologous venoms.These results suggest that different epitopes are present in the protein responsible for the dermonecrotic activity of Loxosceles venoms, and confirm the participation of other venom components during the local reaction process. This study also confirms the importance of antibodies for neutralization of dermonecrotic activity, even when administered some hours after envenoming, and emphasizes the differences of composition and toxicity of medically important Loxosceles venoms. These findings must be considered in order to improve loxoscelism immunotherapy.
Guilherme
P
P
Laboratório de Imunopatologia, Instituto Butantan, Av. Vital Brazil 1500, 05503-900, SP, São Paulo, Brazil.
Fernandes
I
I
Barbaro
K C
KC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000494:pharmacology*; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry; D007120:Immunochemistry; D007928:Lethal Dose 50; D051379:Mice; D008807:Mice, Inbred BALB C; D009336:Necrosis; D012867:Skin / Q000473:pathology*; D013045:Species Specificity; D013111:Spider Venoms / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000633:toxicity*; D013112:Spiders / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D013111:Spider Venoms