Schneider F S, de Almeida Lima S, Reis de Ávila G, Castro K L, Guerra-Duarte C, Sanchez E F, Nguyen C, Granier C, Molina F, Chávez-Olortegui C
Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil; Sys2Diag, FRE 3690, CNRS Alcediag, Languedoc-Roussillon, Montpellier, France.
Departamento de Bioquímica e Imunologia, Instituto Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Vaccine. 2016 Mar 29;34(14):1680-7. doi: 10.1016/j.vaccine.2016.02.035. Epub 2016 Feb 23.
Atroxlysin-I (Atr-I) is a hemorrhagic snake venom metalloproteinase (SVMP) from Bothrops atrox venom, the snake responsible for the majority of bites in the north region of South America. SVMPs like Atr-I produce toxic effects in victims including hemorrhage, inflammation, necrosis and blood coagulation deficiency. Mapping of B-cell epitopes in SVMPs might result in the identification of non-toxic molecules capable of inducing neutralizing antibodies and improving the anti-venom therapy. Here, using the SPOT-synthesis technique we identified two epitopes located in the N-ter region of Atr-I (AtrEp1-(22)YNGNSDKIRRRIHQM(36); and AtrEp2-(55)GVEIWSNKDLINVQ(68)). Based on the sequence of AtrEp1 and AtrEp2 a third peptide named Atr-I biepitope (AtrBiEp) was designed and synthesized ((23)NGNSDKIRRRIH(34)GG(55)GVEIWSNKDLINVQ(68)). AtrBiEp was used to immunize BALB/c mice. Anti-AtrBiEp serum cross-reacted against Atr-I in western blot and was able to fully neutralize the hemorrhagic activity of Atr-I. Our results provide a rational basis for the identification of neutralizing epitopes on Atr-I snake venom toxin and show that the use of synthetic peptides could improve the generation of immuno-therapeutics.
阿卓溶素-I（Atr-I）是一种来自矛头蝮蛇毒液的出血性蛇毒金属蛋白酶（SVMP），矛头蝮蛇是南美洲北部地区大多数咬伤事件的元凶。像Atr-I这样的蛇毒金属蛋白酶会对受害者产生毒性作用，包括出血、炎症、坏死和凝血功能缺陷。绘制蛇毒金属蛋白酶中的B细胞表位图谱可能会鉴定出能够诱导中和抗体并改善抗蛇毒血清疗法的无毒分子。在此，我们使用SPOT合成技术鉴定了位于Atr-I N端区域的两个表位（AtrEp1-(22)YNGNSDKIRRRIHQM(36)；以及AtrEp2-(55)GVEIWSNKDLINVQ(68)）。基于AtrEp1和AtrEp2的序列，设计并合成了第三个名为Atr-I双表位（AtrBiEp）的肽((23)NGNSDKIRRRIH(34)GG(55)GVEIWSNKDLINVQ(68))。AtrBiEp用于免疫BALB/c小鼠。抗AtrBiEp血清在蛋白质印迹中与Atr-I发生交叉反应，并且能够完全中和Atr-I的出血活性。我们的结果为鉴定Atr-I蛇毒毒素上的中和表位提供了合理依据，并表明使用合成肽可以改善免疫治疗药物的产生。
