Immunological characterization of the subunits of type A botulinum neurotoxin and different components of its associated proteins
Immunological characterization of the subunits of type A botulinum neurotoxin and different components of its associated proteins
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DOI：
10.1016/j.toxicon.2009.01.017
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发表时间：
2009-05-01
期刊：
影响因子：
2.8
通讯作者：
Singh, Bal Ram
中科院分区：
文献类型：
--
作者：
Kukreja, Roshan;Chang, Tzuu-Wang;Singh, Bal Ram
肉毒杆菌神经毒素(BoNTs)是由7种结构相似但抗基因差异的蛋白质组成的家族，这些蛋白质由不同的肉毒杆菌菌株产生。A型肉毒杆菌神经毒素(BONT/A)与包括血凝素(HN-33)在内的6种神经毒素相关蛋白(NAP)通过一组基因的多顺反子表达而形成复合体(BONT/AC)。NAPs的存在显著增强了神经毒素的口服毒性。HN-33构成了BONT/AC中NAPs的最大部分，并强烈保护BONT/A免受胃肠道蛋白酶的影响，其复杂形式的BONT还被用于治疗和美容应用，以治疗几种神经肌肉疾病。本研究用酶联免疫吸附试验(ELISA)检测了纯化的和复杂形式的BoNT/A、神经毒素相关蛋白和HN-33的免疫反应性。与纯化的神经毒素相比，针对整个复合体的抗体与复合体的反应性提高了60倍，与HN-33和NAPS的反应性提高了35倍，表明NAPS的免疫原性强于纯化的神经毒素，BoNT/AC及其相关蛋白具有更高的诱导宿主免疫应答的潜力。这一观察还表明，HN-33和其他NAP可能被用作开发肉毒杆菌中毒疫苗的佐剂，并可能成为肉毒杆菌诊断的良好替代品。BoNT/AC和BoNT/A与抗BoNT/A抗体的ELISA结合曲线表明，纯化和复杂形式的BoNT/A具有相同的免疫原性，其免疫原性是BoNT/A轻链和重链的2.5倍。我们还发现了一种新的蛋白质，一种内膜类似物，存在于复杂的BONT/A制剂中，显示出极高的免疫反应性。(C)2009爱思唯尔有限公司。保留所有权利。
Botulinum neurotoxins (BoNTs) constitute a family of seven structurally similar but anti-genically distinct proteins produced by different strains of Clostridium botulinum. Type A botulinum neurotoxin (BoNT/A) is produced along with 6 neurotoxin associated proteins (NAPs) including hemagglutinin (Hn-33) through polycistronic expression of a clustered group of genes to form a complex (BoNT/AC). The presence of NAPs enhances the oral toxicity of the neurotoxin significantly. Hn-33 makes up the largest fraction of NAPs in BoNT/AC and strongly protects BoNT/A against proteases of the GI tract BoNT in its complex form is also used in therapeutic and cosmetic applications to treat several neuromuscular disorders. In this study immunological reactivity of BoNT/A in its purified and complex forms, neurotoxin associated proteins, and Hn-33 have been examined using enzyme-linked immunosorbent assay (ELISA). Antibodies raised against the whole complex reacted 60 times better with the complex and 35 times better with Hn-33 and NAPs compared to the purified neurotoxin suggesting stronger immunogenicity of NAPs over that of purified neurotoxin and a higher potential of BoNT/AC and its associated proteins to induce host immune response. This observation also suggests that Hn-33 and other NAPs could potentially be employed as adjuvants for development of vaccines against botulism and could be a good surrogate for botulinum diagnostics. ELISA binding curves of BoNT/AC and BoNT/A with antibodies raised against BoNT/A indicate that BoNT/A in its purified and complex forms induces equal immunogenic response and a 2.5-fold higher immunogenic response compared to BoNT/A light and heavy chains. We have also discovered a new protein, an intimin analog, present within the complex preparation of BoNT/A which shows dramatically high immunoreactivity. (C) 2009 Elsevier Ltd. All rights reserved.