B subunit of E-coli enterotoxin as adjuvant and carrier in oral and skin vaccination
B subunit of E-coli enterotoxin as adjuvant and carrier in oral and skin vaccination
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DOI：
10.1016/j.vetimm.2006.03.005
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发表时间：
2006-08-15
影响因子：
1.8
通讯作者：
Pitcovski, J.
中科院分区：
文献类型：
--
作者：
Fingerut, E.;Gutter, B.;Pitcovski, J.
粘膜部位是进入人体的主要自然通道之一。通过抗体刺激局部反应作为系统保护，可以增强非活疫苗的效力，并允许用亚单位疫苗接种而不需要注射。粘膜或皮肤接种需要合适的佐剂和载体。大肠杆菌不耐热肠毒素(LT)及其B亚单位(LTB)已被发现是有效的佐剂。本研究的目的是高效生产和纯化重组LTB(BrLTB)，并检测其佐剂和载体性质。编码LTB的基因被克隆并在大肠杆菌中表达，该产物具有与细胞受体GMI神经节苷脂结合的五聚体形式。建立了一种高效纯化和浓缩BrLTB的一步法。用纯化的brLTB经口免疫和肌肉免疫均可产生较高的抗体效价，可检测到完整的毒素。为了测试其佐剂特性，将brLTB与BSA或重组蛋白(减蛋综合征腺病毒的rKnob)混合，并通过肌肉、口服或经皮给药。在口服免疫组和经皮免疫组中，brLTB的加入显著提高了抗体应答水平，但对注射组的抗体应答无明显影响。与单独接种抗原相比，用另一种重组蛋白(传染性法氏囊病病毒病毒蛋白2)和brLTB免疫并不能提高抗体应答。这些结果表明，brLTB的加入使口服和经皮接种蛋白质抗原成为可能。(C)2006爱思唯尔B.V.保留所有权利。
Mucosal sites are one of the main natural ports of entry into the body. Stimulation of a local response by antibodies as the systemic protection may enhance the efficacy of non-living vaccines, and allow for vaccination by subunit vaccines without the need for injection. Mucosal or skin vaccination necessitates a suitable adjuvant and carrier. Escherichia coli heat-labile enterotoxin (LT) and its B subunit (LTB) have been found to be effective adjuvants. The aim of this study was to efficiently produce and purify recombinant LTB (brLTB), and examine its adjuvant and carrier properties. The gene encoding LTB was cloned and expressed in E. coli, and the product was found to have a pentameric form with the ability to bind the cell receptor, GMI ganglioside. A one-step method for efficient purification and concentration of brLTB was developed. Both oral and intramuscular vaccination with purified brLTB yielded high antibody titers, which detected the whole toxin. In an attempt to test its adjuvant characteristics, brLTB was mixed with either BSA or a recombinant protein (rKnob of egg drop syndrome adenovirus) and delivered intramuscularly, orally or transcutaneously. The addition of brLTB significantly elevated the antibody response in groups vaccinated orally and transcutaneously, but had no influence in injected groups. Vaccination with another recombinant protein, (viral protein 2 of infectious bursa] disease virus) supplemented with brLTB did not elevate the antibody response, as compared to vaccination with the antigen alone. These results demonstrate that the addition of brLTB makes oral and transcutaneous vaccination with protein antigens possible. (c) 2006 Elsevier B.V. All rights reserved.