Comparative immunochemical characteristics of botulinum neurotoxin type A and its associated proteins.
Comparative immunochemical characteristics of botulinum neurotoxin type A and its associated proteins.
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DOI：
10.1016/j.toxicon.2013.06.011
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发表时间：
2013-09
期刊：
影响因子：
--
通讯作者：
Singh BR
中科院分区：
文献类型：
--
作者：
Bryant AM;Cai S;Singh BR
肉毒杆菌菌株分泌神经毒素（BoNT）和一组神经毒素相关蛋白（nap），增强口服毒性，并为神经毒素提供保护，使其免受胃肠道酸性、温度和蛋白酶的影响。nap的一个主要成分是Hn-33，这是一种33kDa蛋白，它也具有蛋白酶抗性并强烈保护BoNT。BoNT/A的复杂形式被用作对抗许多神经肌肉疾病和美容目的的商业治疗配方。对这种制剂的免疫反应可能会阻碍其长期使用；因此，表征相关蛋白的免疫学特性是很重要的。本研究旨在通过一系列竞争性酶联免疫吸附试验（ELISA）了解BoNT/A复合物、BoNT、nap和Hn-33的免疫反应性。结果表明，BoNT/A复合物与复杂抗体的竞争是神经毒素的6倍，这证实了BoNT/A复合物较高的免疫原性确实是与神经毒素复合物相关蛋白的结果。而BoNT/A复合物和Hn-33与Hn-33抗体几乎相同的免疫反应性表明，反应性是由于较高的免疫原性，而不是由于复合物中Hn-33的丰度。ELISA和免疫印迹结果表明，Hn-33主要负责在BoNT/A复合体中引发抗体反应，因此有可能将Hn-33用作肉毒杆菌疫苗开发的佐剂。
Clostridium botulinum strains secrete their neurotoxins (BoNT) along with a group of neurotoxin-associated proteins (NAPs) that enhance the oral toxicity and provide protection to the neurotoxin against acidity, temperature and proteases in the G.I. tract. A major component of NAPs is Hn-33, a 33kDa protein, which is also protease resistant and strongly protects BoNT. The complex form of BoNT/A is used as a commercial therapeutic formulation against many neuromuscular disorders and for cosmetic purposes. Immune response against this formulation could hinder its long-term use; therefore, it is important to characterize the immunological properties of the associated proteins. This study aims to understand the immunological reactivity of BoNT/A complex, BoNT, NAPs, and Hn-33 through a series of competitive enzyme-linked immunosorbent assays (ELISA). The results indicated that BoNT/A complex competed 6 times more with complex antibodies compared to the neurotoxin confirming that the higher immunogenicity of BoNT/A complex was indeed a result of the associated proteins with the neurotoxin complex. While the nearly identical immuno-reactivity of BoNT/A complex and Hn-33 with Hn-33 antibodies indicated that the reactivity was due to the higher immunogenicity not the abundance of Hn-33 in the complex. Both the ELISA and immuno-blot results implied that Hn-33 is primarily responsible for eliciting the antibody response in BoNT/A complex, therefore it may be possible to employ Hn-33 as an adjuvant for development of vaccines against botulism.
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