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矛头蝮蛇免疫原组和毒液组学:一种研究蛇毒分子免疫学的蛋白质组学方法。

Immunome and venome of Bothrops jararacussu: a proteomic approach to study the molecular immunology of snake toxins.

机构信息

Instituto de Bioquímica Médica, Programa de Biologia Estrutural, Universidade Federal do Rio de Janeiro 21941-590, Brazil.

出版信息

Toxicon. 2010 Jun 15;55(7):1222-35. doi: 10.1016/j.toxicon.2009.12.018. Epub 2010 Jan 6.

Abstract

A combination of anti-bothropic and anti-crotalic sera has been reported to be more effective in neutralizing the effects of Bothrops jararacussu venom than anti-bothropic serum alone. The role of proteins from B. jararacussu venom in the horse immune response was evaluated via the analysis of cross-reactivity with homologous and heterologous sera. Many of the proteins in B. jararacussu venom were identified via 2D gel electrophoresis. Western blots revealed that anti-jararacussu showed higher reactivity to l-aminoxidase (LAOs) and snake venom metalloproteinase, (SVMPs) and weaker reactivity towards Snake venom serine proteases (SVSPs), PLA(2), C-type lectin and cysteine-rich proteins. Anti-jararaca preferentially recognized LAOs, SVMPs and SVSPs. Both of these sera failed to recognize low-molecular weight proteins. Anti-crotalic serum clearly recognized LAOs, C-type lectin, SVSP, cysteine-rich proteins, SVMP and Asp49-PLA(2). The cross-reactivity with anti-PLA(2) revealed the immunoreactivity of these antibodies to proteins with molecular masses in a range that is poorly recognized by other studied anti-sera. Our results suggest that the contribution of anti-crotalic serum to the neutralization of B. jararacussu by may be due to its cross-reactivity with proteins such as C-type lectins, SVSPs, Asp49-PLA(2). These results also reinforce the importance of neutralizing the highly toxic proteins inclusive those with low immunogenicity in commercial antivenom production to obtain a highly protective serum against snake venoms.

摘要

已报道称,抗矛头蝮蛇血清和抗响尾蛇血清的混合物在中和矛头蝮蛇(Bothrops jararacussu)毒液的作用方面比单独使用抗矛头蝮蛇血清更有效。通过分析同源和异源血清的交叉反应,评估了矛头蝮蛇毒液中的蛋白质在马的免疫反应中的作用。通过 2D 凝胶电泳鉴定了许多矛头蝮蛇毒液中的蛋白质。Western blot 显示抗矛头蝮蛇血清对 l-氨基氧化酶(LAOs)和蛇毒金属蛋白酶(SVMPs)表现出更高的反应性,对蛇毒丝氨酸蛋白酶(SVSPs)、PLA(2)、C 型凝集素和富含半胱氨酸的蛋白质表现出较弱的反应性。抗矛头蝮蛇血清优先识别 LAOs、SVMPs 和 SVSPs。这两种血清都无法识别低分子量蛋白质。抗响尾蛇血清清楚地识别 LAOs、C 型凝集素、SVSP、富含半胱氨酸的蛋白质、SVMP 和 Asp49-PLA(2)。与抗 PLA(2)的交叉反应显示这些抗体对分子量在其他研究抗血清难以识别范围内的蛋白质具有免疫反应性。我们的结果表明,抗响尾蛇血清对 B. jararacussu 的中和作用可能是由于其与 C 型凝集素、SVSPs、Asp49-PLA(2)等蛋白质的交叉反应所致。这些结果还强调了在商业抗蛇毒血清生产中中和包括那些免疫原性低的高度毒性蛋白质的重要性,以获得对蛇毒具有高度保护作用的血清。

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