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针对莫桑比克主要蛇类群体的马源IgG抗蛇毒血清的研制。

Development of Equine IgG Antivenoms against Major Snake Groups in Mozambique.

作者信息

Guidolin Felipe Raimondi, Caricati Celso Pereira, Marcelino José Roberto, da Silva Wilmar Dias

机构信息

Laboratório Especial Piloto de Pesquisa e Desenvolvimento de Imunobiológicos Veterinários (LEPDIV), Instituto Butantan, São Paulo, Brazil.

Divisão de Desenvolvimento Tecnológico e Produção, Soros Hiperimunes, Instituto Butantan, São Paulo, Brazil.

出版信息

PLoS Negl Trop Dis. 2016 Jan 5;10(1):e0004325. doi: 10.1371/journal.pntd.0004325. eCollection 2016 Jan.

Abstract

BACKGROUND

Snake envenoming is a significant public health problem in underdeveloped and developing countries. In sub-Saharan Africa, it is estimated that 90,000-400,000 envenomations occur each year, resulting in 3,500-32,000 deaths. Envenomings are caused by snakes from the Viperidae (Bitis spp. and Echis spp.) and Elapidae (Naja spp. and Dendroaspis spp.) families. The African continent has been suffering from a severe antivenom crisis and current antivenom production is only sufficient to treat 25% of snakebite cases. Our aim is to develop high-quality antivenoms against the main snake species found in Mozambique.

METHODS

Adult horses primed with the indicated venoms were divided into 5 groups (B. arietans; B. nasicornis + B. rhinoceros; N. melanoleuca; N. mossambica; N. annulifera + D. polylepis + D. angusticeps) and reimmunized two times for antivenom production. Blood was collected, and plasma was separated and subjected to antibody purification using caprylic acid. Plasmas and antivenoms were subject to titration, affinity determination, cross-recognition assays and in vivo venom lethality neutralization. A commercial anti-Crotalic antivenom was used for comparison.

RESULTS

The purified antivenoms exhibited high titers against B. arietans, B. nasicornis and B. rhinoceros (5.18 x 106, 3.60 x 106 and 3.50 x 106 U-E/mL, respectively) and N. melanoleuca, N. mossambica and N. annulifera (7.41 x 106, 3.07 x 106 and 2.60 x 106 U-E/mL, respectively), but lower titers against the D. angusticeps and D. polylepis (1.87 x 106 and 1.67 x 106 U-E/mL). All the groups, except anti-N. melanoleuca, showed significant differences from the anti-Crotalic antivenom (7.55 x 106 U-E/mL). The affinity index of all the groups was high, ranging from 31% to 45%. Cross-recognition assays showed the recognition of proteins with similar molecular weight in the venoms and may indicate the possibility of paraspecific neutralization. The three monospecific antivenoms were able to provide in vivo protection.

CONCLUSION

Our results indicate that the anti-Bitis and anti-Naja antivenoms developed would be useful for treating snakebite envenomations in Mozambique, although their effectiveness should to be increased. We propose instead the development of monospecific antivenoms, which would serve as the basis for two polyvalent antivenoms, the anti-Bitis and anti-Elapidae. Polyvalent antivenoms represent an increase in treatment quality, as they have a wider range of application and are easier to distribute and administer to snake envenoming victims.

摘要

背景

蛇咬伤在欠发达国家和发展中国家是一个重大的公共卫生问题。在撒哈拉以南非洲地区,据估计每年发生90,000 - 400,000次蛇咬伤事件,导致3,500 - 32,000人死亡。蛇咬伤是由蝰蛇科(锯鳞蝰属和夜蝰属)和眼镜蛇科(眼镜蛇属和树眼镜蛇属)的蛇类造成的。非洲大陆一直遭受严重的抗蛇毒血清危机,目前的抗蛇毒血清产量仅足以治疗25%的蛇咬伤病例。我们的目标是开发针对莫桑比克主要蛇种的高质量抗蛇毒血清。

方法

用指定毒液对成年马进行初免,分为5组(多带带矛头蝮;角蝰 + 犀咝蝰;黑曼巴;莫桑比克射毒眼镜蛇;环纹眼镜蛇 + 多鳞树眼镜蛇 + 非洲树眼镜蛇),并再次免疫两次以生产抗蛇毒血清。采集血液,分离血浆,并用辛酸进行抗体纯化。对血浆和抗蛇毒血清进行滴定、亲和力测定、交叉识别试验以及体内毒液致死性中和试验。使用一种商业抗响尾蛇毒血清作为对照。

结果

纯化后的抗蛇毒血清对多带带矛头蝮、角蝰和犀咝蝰表现出高滴度(分别为5.18×10⁶、3.60×10⁶和3.50×10⁶ U-E/mL),对黑曼巴、莫桑比克射毒眼镜蛇和环纹眼镜蛇也有高滴度(分别为7.41×10⁶、3.07×10⁶和2.60×10⁶ U-E/mL),但对非洲树眼镜蛇和多鳞树眼镜蛇的滴度较低(分别为1.87×10⁶和1.67×10⁶ U-E/mL)。除抗黑曼巴血清组外,所有组与抗响尾蛇毒血清(7.55×10⁶ U-E/mL)相比均有显著差异。所有组的亲和指数都很高,范围从31%到45%。交叉识别试验表明能识别毒液中分子量相似的蛋白质,这可能表明存在异特异性中和的可能性。三种单特异性抗蛇毒血清能够提供体内保护。

结论

我们的结果表明,所开发的抗锯鳞蝰和抗眼镜蛇抗蛇毒血清虽有效性有待提高,但对治疗莫桑比克的蛇咬伤中毒将是有用的。我们建议改为开发单特异性抗蛇毒血清,这将作为两种多价抗蛇毒血清——抗锯鳞蝰和抗眼镜蛇科抗蛇毒血清的基础。多价抗蛇毒血清代表了治疗质量的提高,因为它们具有更广泛的应用范围,并且更易于分发给蛇咬伤中毒受害者并进行给药。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b6c9/4701360/c02715361f93/pntd.0004325.g001.jpg

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