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Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS

Outer membrane particles from Gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. We previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (GMMA), from S...

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Autores principales: Rossi, Omar, Pesce, Isabella, Giannelli, Carlo, Aprea, Susanna, Caboni, Mariaelena, Citiulo, Francesco, Valentini, Sara, Ferlenghi, Ilaria, MacLennan, Calman Alexander, D'Oro, Ugo, Saul, Allan, Gerke, Christiane
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4155660/
https://www.ncbi.nlm.nih.gov/pubmed/25023285
http://dx.doi.org/10.1074/jbc.M114.566570
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author Rossi, Omar
Pesce, Isabella
Giannelli, Carlo
Aprea, Susanna
Caboni, Mariaelena
Citiulo, Francesco
Valentini, Sara
Ferlenghi, Ilaria
MacLennan, Calman Alexander
D'Oro, Ugo
Saul, Allan
Gerke, Christiane
author_facet Rossi, Omar
Pesce, Isabella
Giannelli, Carlo
Aprea, Susanna
Caboni, Mariaelena
Citiulo, Francesco
Valentini, Sara
Ferlenghi, Ilaria
MacLennan, Calman Alexander
D'Oro, Ugo
Saul, Allan
Gerke, Christiane
author_sort Rossi, Omar
collection PubMed
description Outer membrane particles from Gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. We previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (GMMA), from Shigella. As GMMA are derived from the outer membrane, they contain immunostimulatory components, especially lipopolysaccharide (LPS). We examined ways of reducing their reactogenicity by modifying lipid A, the endotoxic part of LPS, through deletion of late acyltransferase genes, msbB or htrB, in GMMA-producing Shigella sonnei and Shigella flexneri strains. GMMA with resulting penta-acylated lipid A from the msbB mutants showed a 600-fold reduced ability, and GMMA from the S. sonnei ΔhtrB mutant showed a 60,000-fold reduced ability compared with GMMA with wild-type lipid A to stimulate human Toll-like receptor 4 (TLR4) in a reporter cell line. In human peripheral blood mononuclear cells, GMMA with penta-acylated lipid A showed a marked reduction in induction of inflammatory cytokines (S. sonnei ΔhtrB, 800-fold; ΔmsbB mutants, 300-fold). We found that the residual activity of these GMMA is largely due to non-lipid A-related TLR2 activation. In contrast, in the S. flexneri ΔhtrB mutant, a compensatory lipid A palmitoleoylation resulted in GMMA with hexa-acylated lipid A with ∼10-fold higher activity to stimulate peripheral blood mononuclear cells than GMMA with penta-acylated lipid A, mostly due to retained TLR4 activity. Thus, for use as vaccines, GMMA will likely require lipid A penta-acylation. The results identify the relative contributions of TLR4 and TLR2 activation by GMMA, which need to be taken into consideration for GMMA vaccine development.
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spelling pubmed-41556602014-09-05 Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS Rossi, Omar Pesce, Isabella Giannelli, Carlo Aprea, Susanna Caboni, Mariaelena Citiulo, Francesco Valentini, Sara Ferlenghi, Ilaria MacLennan, Calman Alexander D'Oro, Ugo Saul, Allan Gerke, Christiane J Biol Chem Microbiology Outer membrane particles from Gram-negative bacteria are attractive vaccine candidates as they present surface antigens in their natural context. We previously developed a high yield production process for genetically derived particles, called generalized modules for membrane antigens (GMMA), from Shigella. As GMMA are derived from the outer membrane, they contain immunostimulatory components, especially lipopolysaccharide (LPS). We examined ways of reducing their reactogenicity by modifying lipid A, the endotoxic part of LPS, through deletion of late acyltransferase genes, msbB or htrB, in GMMA-producing Shigella sonnei and Shigella flexneri strains. GMMA with resulting penta-acylated lipid A from the msbB mutants showed a 600-fold reduced ability, and GMMA from the S. sonnei ΔhtrB mutant showed a 60,000-fold reduced ability compared with GMMA with wild-type lipid A to stimulate human Toll-like receptor 4 (TLR4) in a reporter cell line. In human peripheral blood mononuclear cells, GMMA with penta-acylated lipid A showed a marked reduction in induction of inflammatory cytokines (S. sonnei ΔhtrB, 800-fold; ΔmsbB mutants, 300-fold). We found that the residual activity of these GMMA is largely due to non-lipid A-related TLR2 activation. In contrast, in the S. flexneri ΔhtrB mutant, a compensatory lipid A palmitoleoylation resulted in GMMA with hexa-acylated lipid A with ∼10-fold higher activity to stimulate peripheral blood mononuclear cells than GMMA with penta-acylated lipid A, mostly due to retained TLR4 activity. Thus, for use as vaccines, GMMA will likely require lipid A penta-acylation. The results identify the relative contributions of TLR4 and TLR2 activation by GMMA, which need to be taken into consideration for GMMA vaccine development. American Society for Biochemistry and Molecular Biology 2014-09-05 2014-07-14 /pmc/articles/PMC4155660/ /pubmed/25023285 http://dx.doi.org/10.1074/jbc.M114.566570 Text en © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/) applies to Author Choice Articles
spellingShingle Microbiology
Rossi, Omar
Pesce, Isabella
Giannelli, Carlo
Aprea, Susanna
Caboni, Mariaelena
Citiulo, Francesco
Valentini, Sara
Ferlenghi, Ilaria
MacLennan, Calman Alexander
D'Oro, Ugo
Saul, Allan
Gerke, Christiane
Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS
title Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS
title_full Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS
title_fullStr Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS
title_full_unstemmed Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS
title_short Modulation of Endotoxicity of Shigella Generalized Modules for Membrane Antigens (GMMA) by Genetic Lipid A Modifications: RELATIVE ACTIVATION OF TLR4 AND TLR2 PATHWAYS IN DIFFERENT MUTANTS
title_sort modulation of endotoxicity of shigella generalized modules for membrane antigens (gmma) by genetic lipid a modifications: relative activation of tlr4 and tlr2 pathways in different mutants
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4155660/
https://www.ncbi.nlm.nih.gov/pubmed/25023285
http://dx.doi.org/10.1074/jbc.M114.566570
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