Centro de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, 30510-010, Belo Horizonte, MG, Brazil.
Mestrado Profissional em Biotecnologia e Gestão da Inovação, Centro Universitário de Sete Lagoas, Sete Lagoas, 35701-242, Minas Gerais, Brazil.
Toxicon. 2019 Sep;167:134-143. doi: 10.1016/j.toxicon.2019.06.019. Epub 2019 Jun 14.
Hyaluronidases (HYALs) are enzymes ubiquitously found in venoms from diverse animals and seem to be related to venom spreading. HYAL activity might be important to Tityus spp. envenoming, since anti-Tityus serrulatus HYAL (TsHYAL) rabbit antibodies neutralize T. serrulatus venom (TsV) lethality. The present work aimed to verify and compare HYAL activity of venoms from other Brazilian Tityus spp. (Tityus bahiensis, Tityus stigmurus and Tityus obscurus) and to test whether anti-TsHYAL antibodies and Brazilian horse therapeutic scorpion antivenom (produced by Fundação Ezequiel Dias (FUNED), Butantan and Vital Brazil Institutes) can recognize and inhibit HYAL activity from these venoms. In ELISA assays, anti-TsHYAL and scorpion antivenoms recognized T. serrulatus, T. bahiensis and T. stigmurus venoms, however, they demonstrated weaker reaction with T. obscurus, which was also observed in Western blotting assay. Epitope mapping by SPOT assay revealed different binding patterns for each antivenom. The assay showed a weaker binding of scorpion antivenom produced by FUNED to peptides recognized by anti-TsHYAL antibodies. Anti-TsHYAL antibodies and antivenoms produced by Butantan and Vital Brazil institutes inhibited HYAL activity of all tested venoms in vitro, whereas FUNED antivenom did not show the same property. These results call attention to the importance of hyaluronidase inhibition, that can aid the improvement of antivenom production.
透明质酸酶（HYALs）广泛存在于各种动物的毒液中，似乎与毒液扩散有关。HYAL 活性可能对 Tityus spp. 的中毒很重要，因为抗 Tityus serrulatus HYAL（TsHYAL）兔抗体可中和 T. serrulatus 毒液（TsV）的致死性。本工作旨在验证和比较来自其他巴西 Tityus spp.（Tityus bahiensis、Tityus stigmurus 和 Tityus obscurus）的毒液中的 HYAL 活性，并测试抗 TsHYAL 抗体和巴西马治疗性蝎抗毒液（由 Fundação Ezequiel Dias（FUNED）、Butantan 和 Vital Brazil 研究所生产）是否能识别和抑制这些毒液中的 HYAL 活性。在 ELISA 测定中，抗 TsHYAL 和蝎抗毒液识别 T. serrulatus、T. bahiensis 和 T. stigmurus 毒液，但与 T. obscurus 的反应较弱，Western blot 测定也观察到了同样的情况。斑点试验的表位作图显示了每种抗毒液的不同结合模式。该试验显示 FUNED 生产的蝎抗毒液与抗 TsHYAL 抗体识别的肽的结合较弱。抗 TsHYAL 抗体和由 Butantan 和 Vital Brazil 研究所生产的抗毒液在体外抑制了所有测试毒液的 HYAL 活性，而 FUNED 抗毒液则没有表现出相同的特性。这些结果表明 HYAL 抑制的重要性，这有助于提高抗毒液的生产。
