A Novel High-Potency Tetanus Vaccine

ABSTRACT Chemically inactivated tetanus toxoid (CITT) is clinically effective and widely used. However, CITT is a crude nonmalleable vaccine that contains hundreds of Clostridium tetani proteins, and the active component is present in variable and sometimes minor percentages of vaccine mass. Recombi...

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Autores principales:	Amanda Przedpelski, William H. Tepp, Sabine Pellett, Eric A. Johnson, Joseph T. Barbieri
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2020
Materias:	DNA recombination Escherichia coli immune response immunization tetanus toxin Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6beab9842e624c3b89a53789ea92360a
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spelling	oai:doaj.org-article:6beab9842e624c3b89a53789ea92360a2021-11-15T15:56:43ZA Novel High-Potency Tetanus Vaccine10.1128/mBio.01668-202150-7511https://doaj.org/article/6beab9842e624c3b89a53789ea92360a2020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01668-20https://doaj.org/toc/2150-7511ABSTRACT Chemically inactivated tetanus toxoid (CITT) is clinically effective and widely used. However, CITT is a crude nonmalleable vaccine that contains hundreds of Clostridium tetani proteins, and the active component is present in variable and sometimes minor percentages of vaccine mass. Recombinant production of a genetically inactivated tetanus vaccine offers an opportunity to replace and improve the current tetanus vaccine. Previous studies showed the feasibility of engineering full-length tetanus toxin (TT) in Escherichia coli. In the present study, full-length TT was engineered with eight individual amino acid mutations (8MTT) to inactivate catalysis, translocation, and host receptor-binding functions, retaining 99.4% amino acid identity to native tetanus toxin. 8MTT purified as a 150-kDa single-chain protein, which trypsin nicked to a 100-kDa heavy chain and 50-kDa light chain. The 8MTT was not toxic for outbred mice and was >50 million-fold less toxic than native TT. Relative to CITT, 8MTT vaccination elicited a strong immune response and showed good vaccine potency against TT challenge. The strength of the immune response to both vaccines varied among individual outbred mice. These data support 8MTT as a candidate vaccine against tetanus and a malleable candidate conjugate vaccine platform to enhance the immune response to polysaccharides and other macromolecular molecules to facilitate a rapid response to emerging microbial pathogens. IMPORTANCE Chemical inactivation is a clinically effective mechanism to detoxify protein toxins to produce vaccines against microbial infections and to serve as a platform for production of conjugate polysaccharide vaccines. This method is widely used for the production of protein toxin vaccines, including tetanus toxoid. However, chemical modification alters the protein structure with unknown effects on antigenicity. Here, a recombinant full-length tetanus toxin (TT) is engineered with 8 mutations (8MTT) that inactivate three toxin functions: catalysis, translocation, and receptor binding. 8MTT is nontoxic and elicits a potent immune response in outbred mice. 8MTT also represents a malleable platform for the production of conjugate vaccines, which can facilitate a rapid vaccine response against emerging microbial pathogens.Amanda PrzedpelskiWilliam H. TeppSabine PellettEric A. JohnsonJoseph T. BarbieriAmerican Society for MicrobiologyarticleDNA recombinationEscherichia coliimmune responseimmunizationtetanus toxinMicrobiologyQR1-502ENmBio, Vol 11, Iss 4 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	DNA recombination Escherichia coli immune response immunization tetanus toxin Microbiology QR1-502
spellingShingle	DNA recombination Escherichia coli immune response immunization tetanus toxin Microbiology QR1-502 Amanda Przedpelski William H. Tepp Sabine Pellett Eric A. Johnson Joseph T. Barbieri A Novel High-Potency Tetanus Vaccine
description	ABSTRACT Chemically inactivated tetanus toxoid (CITT) is clinically effective and widely used. However, CITT is a crude nonmalleable vaccine that contains hundreds of Clostridium tetani proteins, and the active component is present in variable and sometimes minor percentages of vaccine mass. Recombinant production of a genetically inactivated tetanus vaccine offers an opportunity to replace and improve the current tetanus vaccine. Previous studies showed the feasibility of engineering full-length tetanus toxin (TT) in Escherichia coli. In the present study, full-length TT was engineered with eight individual amino acid mutations (8MTT) to inactivate catalysis, translocation, and host receptor-binding functions, retaining 99.4% amino acid identity to native tetanus toxin. 8MTT purified as a 150-kDa single-chain protein, which trypsin nicked to a 100-kDa heavy chain and 50-kDa light chain. The 8MTT was not toxic for outbred mice and was >50 million-fold less toxic than native TT. Relative to CITT, 8MTT vaccination elicited a strong immune response and showed good vaccine potency against TT challenge. The strength of the immune response to both vaccines varied among individual outbred mice. These data support 8MTT as a candidate vaccine against tetanus and a malleable candidate conjugate vaccine platform to enhance the immune response to polysaccharides and other macromolecular molecules to facilitate a rapid response to emerging microbial pathogens. IMPORTANCE Chemical inactivation is a clinically effective mechanism to detoxify protein toxins to produce vaccines against microbial infections and to serve as a platform for production of conjugate polysaccharide vaccines. This method is widely used for the production of protein toxin vaccines, including tetanus toxoid. However, chemical modification alters the protein structure with unknown effects on antigenicity. Here, a recombinant full-length tetanus toxin (TT) is engineered with 8 mutations (8MTT) that inactivate three toxin functions: catalysis, translocation, and receptor binding. 8MTT is nontoxic and elicits a potent immune response in outbred mice. 8MTT also represents a malleable platform for the production of conjugate vaccines, which can facilitate a rapid vaccine response against emerging microbial pathogens.
format	article
author	Amanda Przedpelski William H. Tepp Sabine Pellett Eric A. Johnson Joseph T. Barbieri
author_facet	Amanda Przedpelski William H. Tepp Sabine Pellett Eric A. Johnson Joseph T. Barbieri
author_sort	Amanda Przedpelski
title	A Novel High-Potency Tetanus Vaccine
title_short	A Novel High-Potency Tetanus Vaccine
title_full	A Novel High-Potency Tetanus Vaccine
title_fullStr	A Novel High-Potency Tetanus Vaccine
title_full_unstemmed	A Novel High-Potency Tetanus Vaccine
title_sort	novel high-potency tetanus vaccine
publisher	American Society for Microbiology
publishDate	2020
url	https://doaj.org/article/6beab9842e624c3b89a53789ea92360a
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A Novel High-Potency Tetanus Vaccine

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