Structural basis of tetanus toxin neutralization by native human monoclonal antibodies

Wang, Yueming; Wu, Changwen; Yu, Jinfang; Lin, Shujian; Liu, Tong; Zan, Lipeng; Li, Nan; Hong, Po; Wang, Xiaoli; Jia, Zhenxing; Li, Jason; Wang, Yao; Zhang, Ming; Yuan, Xiaohui; Li, Chengming; Xu, Wenwen; Zheng, Wenwei; Wang, Xinquan; Liao, Hua‐Xin

doi:10.1016/j.celrep.2021.109070

Cited by 15 publications

(16 citation statements)

References 44 publications

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“…The search for therapeutic anti-tetanus antibodies to replace human immunoglobulin and equine serum has been the subject of recent publications, demonstrating an increased interest in this therapeutic approach [ 25 , 26 , 27 , 28 ]. In common among them is the use of B cells from immunized individuals.…”

Section: Discussionmentioning

confidence: 99%

Characterization of Neutralizing Human Anti-Tetanus Monoclonal Antibodies Produced by Stable Cell Lines

et al. 2022

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Tetanus toxin (TeNT) is produced by C. tetani, a spore-forming bacillus broadly spread in the environment. Although an inexpensive and safe vaccine is available, tetanus persists because of a lack of booster shots and variable responses to vaccines due to immunocompromised status or age-decreased immune surveillance. Tetanus is most prevalent in low- and medium-income countries, where it remains a health problem. Neutralizing monoclonal antibodies (mAbs) can prevent the severity of illness and death caused by C. tetani infection. We identified a panel of mAbs that bind to TeNT, some of which were investigated in a preclinical assay, showing that a trio of mAbs that bind to different sites of TeNT can neutralize the toxin and prevent symptoms and death in mice. We also identified two mAbs that can impair the binding of TeNT to the GT1b ganglioside receptor in neurons. In this work, to generate a series of cell lines, we constructed vectors containing sequences encoding heavy and light constant regions that can receive the paired variable regions resulting from PCRs of human B cells. In this way, we generated stable cell lines for five mAbs and compared and characterized the antibody produced in large quantities, enabling the characterization experiments. We present the results regarding the cell growth and viability in a fed-batch culture, titer measurement, and specific productivity estimation. The affinity of purified mAbs was analyzed by kinetics and under steady-state conditions, as three mAbs could not dissociate from TeNT within 36,000 s. The binding of mAbs to TeNT was confirmed by ELISA and inhibition of toxin binding to GT1b. The use of the mAbs mixture confirmed the individual mAb contribution to inhibition. We also analyzed the binding of mAbs to FcγR by surface plasmon resonance (SPR) and the glycan composition. Molecular docking analyses showed the binding site of an anti-tetanus mAb.

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Section: Discussionmentioning

confidence: 99%

Characterization of Neutralizing Human Anti-Tetanus Monoclonal Antibodies Produced by Stable Cell Lines

et al. 2022

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“…This orients the BoNT molecule while approaching the plasma membrane thus favoring their landing with the HC-C domain and increasing the probability of membrane binding (Fogolari et al 2009 ). The role of PSGs as primary receptors is supported by several experimental evidence (Binz and Rummel 2009 ), and by the fact that the most TeNT- or BoNT-neutralizing monoclonal antibodies sterically prevent PSG binding (Garcia-Rodriguez et al 2007 , 2021 ; Mukherjee et al 2012 ; Lam et al 2018 ; Tomic et al 2021 ; Pirazzini et al 2021 ; Wang et al 2016 , 2021 ; Zhang et al 2021 ; Brier et al 2021 ).…”

Section: Structure and Mechanism Of Action Of Tent And Bontsmentioning

confidence: 98%

“…In the case of tetanus, recent research has focused on human monoclonal antibodies (humAbs) anti-TeNT (Wang et al 2016 , 2021 ; Aliprandini et al 2019 ; Ghotloo et al 2020 ; Minamitani et al 2021 ; Pirazzini et al 2021 ; Zhang et al 2021 ) with the aim of overcoming some clinical problems encountered with the use of the IgG fraction isolated from human polyclonal antisera derived from tetanus toxoid immunized donors (Pirazzini et al 2021 ). The toxin neutralizing humAbs were identified using phage display libraries or by screening antibody producing cells isolated from human individuals that were treated in adult age with the tetanus toxoid vaccine.…”

Section: Toxicology Of Tetanus and Botulinum Neurotoxinsmentioning

confidence: 99%

Toxicology and pharmacology of botulinum and tetanus neurotoxins: an update

2022

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Tetanus and botulinum neurotoxins cause the neuroparalytic syndromes of tetanus and botulism, respectively, by delivering inside different types of neurons, metalloproteases specifically cleaving the SNARE proteins that are essential for the release of neurotransmitters. Research on their mechanism of action is intensively carried out in order to devise improved therapies based on antibodies and chemical drugs. Recently, major results have been obtained with human monoclonal antibodies and with single chain antibodies that have allowed one to neutralize the metalloprotease activity of botulinum neurotoxin type A1 inside neurons. In addition, a method has been devised to induce a rapid molecular evolution of the metalloprotease domain of botulinum neurotoxin followed by selection driven to re-target the metalloprotease activity versus novel targets with respect to the SNARE proteins. At the same time, an intense and wide spectrum clinical research on novel therapeutics based on botulinum neurotoxins is carried out, which are also reviewed here.

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“…Moreover, TT neutralizing antibodies were shown to be mainly directed against TTFC in mouse and human models (100% for mouse models and 75% in a human model) [61]. It is important to note that the many neutralizing antibodies described since the 1980s [62][63][64] cannot individually neutralize TT in vivo. The development of an active and safe monoclonal antibody (mAb) remains a prospect for upcoming years [65].…”

Section: Immunological Properties Against Tetanusmentioning

confidence: 99%

Tetanus Toxin Fragment C: Structure, Drug Discovery Research and Production

et al. 2022

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Tetanus toxoid (TTd) plays an important role in the pharmaceutical world, especially in vaccines. The toxoid is obtained after formaldehyde treatment of the tetanus toxin. In parallel, current emphasis in the drug discovery field is put on producing well-defined and safer drugs, explaining the interest in finding new alternative proteins. The tetanus toxin fragment C (TTFC) has been extensively studied both as a neuroprotective agent for central nervous system disorders owing to its neuronal properties and as a carrier protein in vaccines. Indeed, it is derived from a part of the tetanus toxin and, as such, retains its immunogenic properties without being toxic. Moreover, this fragment has been well characterized, and its entire structure is known. Here, we propose a systematic review of TTFC by providing information about its structural features, its properties and its methods of production. We also describe the large uses of TTFC in the field of drug discovery. TTFC can therefore be considered as an attractive alternative to TTd and remarkably offers a wide range of uses, including as a carrier, delivery vector, conjugate, booster, inducer, and neuroprotector.

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Structural basis of tetanus toxin neutralization by native human monoclonal antibodies

Cited by 15 publications

References 44 publications

Characterization of Neutralizing Human Anti-Tetanus Monoclonal Antibodies Produced by Stable Cell Lines

Characterization of Neutralizing Human Anti-Tetanus Monoclonal Antibodies Produced by Stable Cell Lines

Toxicology and pharmacology of botulinum and tetanus neurotoxins: an update

Tetanus Toxin Fragment C: Structure, Drug Discovery Research and Production

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