Characterization of the antibody response to the receptor binding domain of botulinum neurotoxin serotypes A and E
Characterization of the antibody response to the receptor binding domain of botulinum neurotoxin serotypes A and E
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DOI：
10.1128/iai.73.10.6998-7005.2005
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发表时间：
2005-10-01
影响因子：
3.1
通讯作者：
Barbieri, JT
中科院分区：
文献类型：
--
作者：
Baldwin, MR;Tepp, WH;Barbieri, JT
肉毒杆菌神经毒素（BoNTs）是对人类毒性最大的蛋白质。目前针对BoNT中毒的梭菌衍生疫苗在生产和可及性方面存在局限性。建立了在大肠杆菌中表达A、E血清型BoNT可溶性受体结合域（重链受体[HCR]）的条件。用经典A-Hall菌株（ATCC 3502） （BoNT/A1）的重组HCR/A1 （rHCR/A1）和E- Beluga血清型BoNT （BoNT/E- b）的rHCR/E免疫小鼠和家兔的血清可以中和同源的BoNT血清型，但在交叉识别和交叉保护方面存在差异。酶联免疫吸附试验和Western blotting结果显示，α - rhcr /A1识别的抗原表位位于HCR/A和HCR/E的C端，而α - rhcr /E识别的抗原表位位于HCR蛋白的N端或N端与C端交界面。α - rhcr /E- b血清对BoNT/A1具有可检测的中和能力，而α - rhcr /A1不能中和BoNT/E。rHCR/A是对BoNT/A1和京都F婴儿株（BoNT/A2）有效的免疫原，但对BoNT阿拉斯加E血清型（BoNT/E-A）无效，而rHCR/E- b中和BoNT/E-A，并在过度免疫条件下对BoNT/A1和BoNT/A2有保护作用。rHCR/A1对BoNT/A1和BoNT/A2的保护作用以及rHCR/EB对BoNT/EA的保护作用表明，受体结合域免疫可在BoNT亚型中引起保护作用。rHCR/E高免疫对BoNT/A的保护作用表明，E和A血清型之间存在共同的中和表位。这些结果表明，受体结合域亚单位疫苗对BoNT的血清型变体具有保护作用。
Clostridium botulinum neurotoxins (BoNTs) are the most toxic proteins for humans. The current clostridial-derived vaccines against BoNT intoxication have limitations including production and accessibility. Conditions were established to express the soluble receptor binding domain (heavy-chain receptor [HCR]) of BoNT serotypes A and E in Escherichia coli. Sera isolated from mice and rabbits immunized with recombinant HCR/A1 (rHCR/A1) from the classical type A-Hall strain (ATCC 3502) (BoNT/A1) and rHCR/E from BoNT serotype E Beluga (BoNT/E-B) neutralized the homologous serotype of BoNT but displayed differences in cross-recognition and cross-protection. Enzyme-linked immunosorbent assay and Western blotting showed that alpha-rHCR/A1 recognized epitopes within the C terminus of the HCR/A and HCR/E, while alpha-rHCR/E recognized epitopes within the N terminus or interface between the N and C termini of the HCR proteins. alpha-rHCR/E-B sera possessed detectable neutralizing capacity for BoNT/A1, while alpha-rHCR/A1 did not neutralize BoNT/E. rHCR/A was an effective immunogen against BoNT/A1 and the Kyoto F infant strain (BoNT/A2), but not BoNT serotype E Alaska (BoNT/E-A), while rHCR/E-B neutralized BoNT/E-A, and under hyperimmunization conditions protected against BoNT/A1 and BoNT/A2. The protection elicited by rHCR/A1 to BoNT/A1 and BoNT/A2 and by rHCR/EB to BoNT/EA indicate that immunization with receptor binding domains elicit protection within sub-serotypes of BoNT. The protection elicited by hyperimmunization with rHCR/E against BoNT/A suggests the presence of common neutralizing epitopes between the serotypes E and A. These results show that a receptor binding domain subunit vaccine protects against serotype variants of BoNTs.