<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant

ABSTRACT Recombinant attenuated Salmonella enterica serovar Typhimurium strains are believed to act as powerful live vaccine carriers that are able to elicit protection against various pathogens. Auxotrophic mutations, such as a deletion of aroA, are commonly introduced into such bacteria for attenu...

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Autores principales:	Sebastian Felgner, Michael Frahm, Dino Kocijancic, Manfred Rohde, Denitsa Eckweiler, Agata Bielecka, Emilio Bueno, Felipe Cava, Wolf-Rainer Abraham, Roy Curtiss, Susanne Häussler, Marc Erhardt, Siegfried Weiss
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/b28681446d39498e87e4324eb3ecfd84
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spelling	oai:doaj.org-article:b28681446d39498e87e4324eb3ecfd842021-11-15T15:50:14Z<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant10.1128/mBio.01220-162150-7511https://doaj.org/article/b28681446d39498e87e4324eb3ecfd842016-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01220-16https://doaj.org/toc/2150-7511ABSTRACT Recombinant attenuated Salmonella enterica serovar Typhimurium strains are believed to act as powerful live vaccine carriers that are able to elicit protection against various pathogens. Auxotrophic mutations, such as a deletion of aroA, are commonly introduced into such bacteria for attenuation without incapacitating immunostimulation. In this study, we describe the surprising finding that deletion of aroA dramatically increased the virulence of attenuated Salmonella in mouse models. Mutant bacteria lacking aroA elicited increased levels of the proinflammatory cytokine tumor necrosis factor alpha (TNF-α) after systemic application. A detailed genetic and phenotypic characterization in combination with transcriptomic and metabolic profiling demonstrated that ΔaroA mutants display pleiotropic alterations in cellular physiology and lipid and amino acid metabolism, as well as increased sensitivity to penicillin, complement, and phagocytic uptake. In concert with other immunomodulating mutations, deletion of aroA affected flagellin phase variation and gene expression of the virulence-associated genes arnT and ansB. Finally, ΔaroA strains displayed significantly improved tumor therapeutic activity. These results highlight the importance of a functional shikimate pathway to control homeostatic bacterial physiology. They further highlight the great potential of ΔaroA-attenuated Salmonella for the development of vaccines and cancer therapies with important implications for host-pathogen interactions and translational medicine. IMPORTANCE Recombinant attenuated bacterial vector systems based on genetically engineered Salmonella have been developed as highly potent vaccines. Due to the pathogenic properties of Salmonella, efficient attenuation is required for clinical applications. Since the hallmark study by Hoiseth and Stocker in 1981 (S. K. Hoiseth and B. A. D. Stocker, Nature 291:238–239, 1981, http://dx.doi.org/10.1038/291238a0), the auxotrophic ΔaroA mutation has been generally considered safe and universally used to attenuate bacterial strains. Here, we are presenting the remarkable finding that a deletion of aroA leads to pronounced alterations of gene expression, metabolism, and cellular physiology, which resulted in increased immunogenicity, virulence, and adjuvant potential of Salmonella. These results suggest that the enhanced immunogenicity of aroA-deficient Salmonella strains might be advantageous for optimizing bacterial vaccine carriers and immunotherapy. Accordingly, we demonstrate a superior performance of ΔaroA Salmonella in bacterium-mediated tumor therapy. In addition, the present study highlights the importance of a functional shikimate pathway to sustain bacterial physiology and metabolism.Sebastian FelgnerMichael FrahmDino KocijancicManfred RohdeDenitsa EckweilerAgata BieleckaEmilio BuenoFelipe CavaWolf-Rainer AbrahamRoy CurtissSusanne HäusslerMarc ErhardtSiegfried WeissAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 5 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Sebastian Felgner Michael Frahm Dino Kocijancic Manfred Rohde Denitsa Eckweiler Agata Bielecka Emilio Bueno Felipe Cava Wolf-Rainer Abraham Roy Curtiss Susanne Häussler Marc Erhardt Siegfried Weiss <italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant
description	ABSTRACT Recombinant attenuated Salmonella enterica serovar Typhimurium strains are believed to act as powerful live vaccine carriers that are able to elicit protection against various pathogens. Auxotrophic mutations, such as a deletion of aroA, are commonly introduced into such bacteria for attenuation without incapacitating immunostimulation. In this study, we describe the surprising finding that deletion of aroA dramatically increased the virulence of attenuated Salmonella in mouse models. Mutant bacteria lacking aroA elicited increased levels of the proinflammatory cytokine tumor necrosis factor alpha (TNF-α) after systemic application. A detailed genetic and phenotypic characterization in combination with transcriptomic and metabolic profiling demonstrated that ΔaroA mutants display pleiotropic alterations in cellular physiology and lipid and amino acid metabolism, as well as increased sensitivity to penicillin, complement, and phagocytic uptake. In concert with other immunomodulating mutations, deletion of aroA affected flagellin phase variation and gene expression of the virulence-associated genes arnT and ansB. Finally, ΔaroA strains displayed significantly improved tumor therapeutic activity. These results highlight the importance of a functional shikimate pathway to control homeostatic bacterial physiology. They further highlight the great potential of ΔaroA-attenuated Salmonella for the development of vaccines and cancer therapies with important implications for host-pathogen interactions and translational medicine. IMPORTANCE Recombinant attenuated bacterial vector systems based on genetically engineered Salmonella have been developed as highly potent vaccines. Due to the pathogenic properties of Salmonella, efficient attenuation is required for clinical applications. Since the hallmark study by Hoiseth and Stocker in 1981 (S. K. Hoiseth and B. A. D. Stocker, Nature 291:238–239, 1981, http://dx.doi.org/10.1038/291238a0), the auxotrophic ΔaroA mutation has been generally considered safe and universally used to attenuate bacterial strains. Here, we are presenting the remarkable finding that a deletion of aroA leads to pronounced alterations of gene expression, metabolism, and cellular physiology, which resulted in increased immunogenicity, virulence, and adjuvant potential of Salmonella. These results suggest that the enhanced immunogenicity of aroA-deficient Salmonella strains might be advantageous for optimizing bacterial vaccine carriers and immunotherapy. Accordingly, we demonstrate a superior performance of ΔaroA Salmonella in bacterium-mediated tumor therapy. In addition, the present study highlights the importance of a functional shikimate pathway to sustain bacterial physiology and metabolism.
format	article
author	Sebastian Felgner Michael Frahm Dino Kocijancic Manfred Rohde Denitsa Eckweiler Agata Bielecka Emilio Bueno Felipe Cava Wolf-Rainer Abraham Roy Curtiss Susanne Häussler Marc Erhardt Siegfried Weiss
author_facet	Sebastian Felgner Michael Frahm Dino Kocijancic Manfred Rohde Denitsa Eckweiler Agata Bielecka Emilio Bueno Felipe Cava Wolf-Rainer Abraham Roy Curtiss Susanne Häussler Marc Erhardt Siegfried Weiss
author_sort	Sebastian Felgner
title	<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant
title_short	<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant
title_full	<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant
title_fullStr	<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant
title_full_unstemmed	<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant
title_sort	<italic toggle="yes">aroa</italic>-deficient <named-content content-type="genus-species">salmonella enterica</named-content> serovar typhimurium is more than a metabolically attenuated mutant
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/b28681446d39498e87e4324eb3ecfd84
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<italic toggle="yes">aroA</italic>-Deficient <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Is More Than a Metabolically Attenuated Mutant

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