Development and evaluation of candidate subunit vaccine and novel antitoxin against botulinum neurotoxin serotype E

Shi, Danyang; Liu, Fujia; Mao, Yunyun; Cui, Rong-Tian; Lu, Jian-Sheng; Yu, Yun-Zhou; Dong, Xiaojie; Yang, Zhengming; Sun, Zhiwei; Pang, Xiaobin

doi:10.1080/21645515.2019.1633878

Cited by 8 publications

(12 citation statements)

References 35 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In the last 20 years, the immunological significance of the Hc domain of BoNTs has been the subject of intensive research, leading to important findings [14][15][16][20][21][22][23][24][25]. Our research also indicated that the Hc domain is an effective protective antigen in subunit vaccines for BoNT/A, B, E, and F [26][27][28][29]. However, the immunogenicity of other regions requires further study, such as their protective efficacy and the synergistic effects of multiple epitopes.…”

Section: Discussionmentioning

confidence: 82%

“…To produce recombinant functional fragment antigens of BoNT/E in Escherichia coli, gene fragments of each function fragment (EL, EHN, EL-HN, EHc, EHc-N, and EHc-C, Table 1) were ligated into a prokaryotic expression vector, pTIG-Trx, to produce recombinant expression plasmids. As previously reported [26,36], our laboratory constructed pTIG-Trx prokaryotic expression plasmids encoding the following BoNT/E molecule fragments with His-tag: EL (residues 1-422), EHN (residues 423-840), EL-HN (residues 1-840), EHc-N (residues 841-1063), EHc-C (residues 1056-1252), and EHc (residues 841-1252). The resulting recombinant plasmids were confirmed by sequencing.…”

Section: Production Of Recombinant Functional Fragment Antigens Deriv...mentioning

confidence: 99%

“…A BoNT/E neutralization assay [26] was used to measure the neutralizing potency of the mouse sera. Briefly, serum samples from immunized mice were serially diluted before being mixed with a BoNT/E standard concentration solution (100 LD 50 /mL, 1 mL).…”

Section: Determination Of the Titer Of Neutralizing Antibodies In Mou...mentioning

confidence: 99%

See 2 more Smart Citations

Functional EL-HN Fragment as a Potent Candidate Vaccine for the Prevention of Botulinum Neurotoxin Serotype E

Lu²,

Tan

et al. 2022

Toxins

View full text Add to dashboard Cite

Clostridium botulinum produces botulinum neurotoxin (BoNT), which is the most toxic known protein and the causative agent of human botulism. BoNTs have similar structures and functions, comprising three functional domains: catalytic domain (L), translocation domain (HN), and receptor-binding domain (Hc). In the present study, BoNT/E was selected as a model toxin to further explore the immunological significance of each domain. The EL-HN fragment (L and HN domains of BoNT/E) retained the enzymatic activity without in vivo neurotoxicity. Extensive investigations showed EL-HN functional fragment had the highest protective efficacy and contained some functional neutralizing epitopes. Further experiments demonstrated the EL-HN provided a superior protective effect compared with the EHc or EHc and EL-HN combination. Thus, the EL-HN played an important role in immune protection against BoNT/E and could provide an excellent platform for the design of botulinum vaccines and neutralizing antibodies. The EL-HN has the potential to replace EHc or toxoid as the optimal immunogen for the botulinum vaccine.

show abstract

Section: Discussionmentioning

confidence: 82%

Section: Production Of Recombinant Functional Fragment Antigens Deriv...mentioning

confidence: 99%

See 1 more Smart Citation

Functional EL-HN Fragment as a Potent Candidate Vaccine for the Prevention of Botulinum Neurotoxin Serotype E

Lu²,

Tan

et al. 2022

Toxins

View full text Add to dashboard Cite

show abstract

“…twice with vaccines of different formulation. 20 TBV with Freund’s adjuvant (Sigma-Aldrich, Inc., St Louis, MO, USA) was prepared according the manufacturer’s instructions, in which complete Freund’s adjuvant (F5581) was used for the first immunization and incomplete Freund’s adjuvant (F5506) was used for the second immunization. A formulation without any antigens was used as a negative control.…”

Section: Methodsmentioning

confidence: 99%

“…Sera from mice in different treatment groups were screened for anti-Hc antibody titer by ELISA as previously describe. 20 Sera antibody titers of individual mice were assayed, and the geometric mean titer (GMT) for each group was determined.…”

Section: Methodsmentioning

confidence: 99%

Development and evaluation of a tetravalent botulinum vaccine

Shi¹,

Liu

et al. 2022

Human Vaccines & Immunotherapeutics

Self Cite

View full text Add to dashboard Cite

Botulinum neurotoxins (BoNTs) are the most toxic known proteins. Naturally occurring botulism in humans is caused by botulinum serotypes A, B, E, and F. Vaccination is an effective strategy to prevent botulism. In this study, a tetravalent botulinum vaccine (TBV) that can prevent serotypes A, B, E, and F was developed using the C-terminal receptor-binding domain of BoNT (Hc) as an antigen. To develop a suitable vaccine formulation, in vitro binding experiments of antigens and aluminum adjuvant in different buffers, and in vivo experiments of TBV at different antigen concentrations, were conducted. Our results showed that the optimal vaccine formulation buffer was a pH 6.0 phosphate buffer, and the suitable antigen concentration was 40 or 80 µg/ml of each antigen. A pilot-scale TBV was then prepared and evaluated for immunogenicity and stability. The results showed that TBV could elicit strong protective efficacy against each BoNT in mice, and remain effective after two years of storage at 4ºC, indicating that the preparation was stable and highly effective. Adsorption experiments also showed that the antigens could be well adsorbed by the aluminum adjuvant after 2 years of storage. Our results provide valuable experimental data supporting the development of a tetravalent botulinum vaccine, which is a promising candidate for the prevention of botulinum serotypes A, B, E, and F.

show abstract

Characterization of a novel tetravalent botulism antitoxin based on receptor-binding domain of BoNTs

Shi¹,

Lu²,

Mao³

et al. 2023

Appl Microbiol Biotechnol

Self Cite

View full text Add to dashboard Cite

Botulinum neurotoxin (BoNTs; serotypes A, B, E, and F) cause botulism disease in humans, which could be effectively treated using antitoxins. Herein, we established a novel receptor-binding domain (RBD)-based antitoxin using recombinant C terminal heavy chain (Hc) domains of BoNTs as immunogens. Immunization of horses with these recombinant Hc domains allowed the purification and digestion of IgGs from hyper-immune sera to produce high-quality and high-efficiency monovalent botulism antitoxin F(ab′) 2 against each BoNT (M-BATs). However, these M-BATs could not bind or neutralize other serotypes of BoNTs, and that there were no cross-protective effects among these M-BATs. This suggested the need to prepare tetravalent antitoxins to neutralize the four BoNTs simultaneously. Thus, these M-BATs were formulated into a novel tetravalent botulism antitoxin (T-BAT), in which a 10-ml volume contained 10000 IU of BoNT/A and 5000 IU of BoNT/B, BoNT/E, and BoNT/F antitoxins. The novel antitoxin preparation could prevent and treat the four mixed botulinum neurotoxins simultaneously in vivo, representing strong efficacy in an animal poisoning model. Moreover, these antibodies in T-BAT could bind the RBD, whereas conventional antitoxins based on inactivated toxins mainly bind the light chain or heavy chain translocation domain (HN) and weakly bind the important RBD in current experimental conditions. The high levels of RBD-specific novel antitoxins can efficiently bind the RBD and neutralize natural or recombinant toxins containing this RBD. The findings of the present study experimentally support the use of RBD-specific antitoxins to treat BoNT serotype A, B, E, and F-mediated botulism. This study demonstrated the concept of developing potent novel multivalent antitoxins against all BoNTs or other toxins, using the RBD of these toxins as an alternative antigen to inactivated toxins. Key points • Antitoxins based on the receptor-binding domains of botulinum neurotoxins were made. • Novel antitoxin binds RBD; traditional antitoxin mainly binds light chain or HN domain. • A tetravalent antitoxin could prevent and treat the four mixed neurotoxins in vivo. Supplementary Information The online version contains supplementary material available at 10.1007/s00253-023-12515-2.

show abstract

Development and evaluation of candidate subunit vaccine and novel antitoxin against botulinum neurotoxin serotype E

Cited by 8 publications

References 35 publications

Functional EL-HN Fragment as a Potent Candidate Vaccine for the Prevention of Botulinum Neurotoxin Serotype E

Functional EL-HN Fragment as a Potent Candidate Vaccine for the Prevention of Botulinum Neurotoxin Serotype E

Development and evaluation of a tetravalent botulinum vaccine

Characterization of a novel tetravalent botulism antitoxin based on receptor-binding domain of BoNTs

Contact Info

Product

Resources

About