Abstract
Botulinum neurotoxin (BoNT) is a neurotoxin produced by Clostridium botulinum in an anaerobic environment. BoNT is the most toxic protein among bacteria, animals, plants, and chemical substances reported to date. BoNTs are 150 kDa proteins composed of three major functional domains: catalytic (L domain, 50 kDa), translocation (HN domain, 50 kDa), and receptor-binding (Hc domain, 50 kDa) domains. Most studies have focused on the use of the Hc domain as an antigen because it is capable of generating robust protective immunity and contains some functional neutralizing epitopes. In the present study, we produced and characterized a recombinant L-HN fusion fragment of the parent BoNT/B (BL-HN) composed of L and HN domains with a deletion in the Hc domain (BHc). When the BL-HN protein was expressed in E. coli, it retained its stable structure and antigenicity. As a vaccine antigen, the recombinant BL-HN protein was found to induce sufficient protection against native BoNT/B in a mouse model. The BL-HN subunit vaccine could also induce a strong humoral immune response and generate sufficient neutralizing antibodies in immunized mice. Therefore, BL-HN may retain the native neurotoxin structure and critical epitopes responsible for inducing serum neutralizing antibodies. Studies of the dose-dependent immunoprotective effects further confirmed that the BL-HN antigen could provide potent protective immunity. This finding suggests that BL-HN can play an important role in immune protection against BoNT/B. Therefore, the BL-HN fusion fragment provides an excellent platform for the design of recombinant botulinum vaccines and neutralizing antibodies.
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Abbreviations
- BoNT:
-
Botulinum neurotoxin
- BHc:
-
The Hc domain of BoNT/B
- BL-HN:
-
Recombinant L-HN fusion fragment of the parent BoNT/B composed of L and HN domains
- LD50 :
-
50% Mouse lethal dose
- GMT:
-
Geometric mean titer
- ED50 :
-
The 50% percent effective concentrations
- PBS:
-
Phosphate-buffered saline
- ELISA:
-
Enzyme-linked immunosorbent assay
- TBS:
-
Tris-buffered saline
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YYZ, XQ, and YZX designed the study. LZ and LJS carried out the experiments, and LZ, WR, and LS analyzed the data. LZ and YYZ drafted the manuscript and verified the important intellectual content. All authors critically read and revised the manuscript and approved the final version of the manuscript for submission.
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All animal experiments were approved by the Institution of Animal Care and Use Committee of the Bei**g Institute of Biotechnology and performed in accordance with the ethical guidelines of our institution and the National Institutes of Health guide for the Care and Use of Laboratory Animals.
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Zhen Li, Jian-Sheng Lu, and Shan Liu equally contributed to this work.
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Li, Z., Lu, JS., Liu, S. et al. Recombinant L-HN Fusion Antigen Derived from the L and HN Domains of Botulinum Neurotoxin B Stimulates a Protective Antibody Response Against Active Neurotoxin. Neurotox Res 39, 1044–1053 (2021). https://doi.org/10.1007/s12640-021-00337-x
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DOI: https://doi.org/10.1007/s12640-021-00337-x