Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery

ABSTRACT Adjuvant properties of bacterial cell wall components like MPLA (monophosphoryl lipid A) are well described and have gained FDA approval for use in vaccines such as Cervarix. MPLA is the product of chemically modified lipooligosaccharide (LOS), altered to diminish toxic proinflammatory effe...

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Autores principales:	Kelsey A. Gregg, Erin Harberts, Francesca M. Gardner, Mark R. Pelletier, Corinne Cayatte, Li Yu, Michael P. McCarthy, Jason D. Marshall, Robert K. Ernst
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2017
Materias:	TLR4 adjuvants immunomodulation innate immunity lipid A lipopolysaccharide Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9e94f7b9ae6c480f9062857e69a6fc14
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spelling	oai:doaj.org-article:9e94f7b9ae6c480f9062857e69a6fc142021-11-15T15:51:28ZRationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery10.1128/mBio.00492-172150-7511https://doaj.org/article/9e94f7b9ae6c480f9062857e69a6fc142017-07-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00492-17https://doaj.org/toc/2150-7511ABSTRACT Adjuvant properties of bacterial cell wall components like MPLA (monophosphoryl lipid A) are well described and have gained FDA approval for use in vaccines such as Cervarix. MPLA is the product of chemically modified lipooligosaccharide (LOS), altered to diminish toxic proinflammatory effects while retaining adequate immunogenicity. Despite the virtually unlimited number of potential sources among bacterial strains, the number of useable compounds within this promising class of adjuvants are few. We have developed bacterial enzymatic combinatorial chemistry (BECC) as a method to generate rationally designed, functionally diverse lipid A. BECC removes endogenous or introduces exogenous lipid A-modifying enzymes to bacteria, effectively reprogramming the lipid A biosynthetic pathway. In this study, BECC is applied within an avirulent strain of Yersinia pestis to develop structurally distinct LOS molecules that elicit differential Toll-like receptor 4 (TLR4) activation. Using reporter cell lines that measure NF-κB activation, BECC-derived molecules were screened for the ability to induce a lower proinflammatory response than Escherichia coli LOS. Their structures exhibit varied, dose-dependent, TLR4-driven NF-κB activation with both human and mouse TLR4 complexes. Additional cytokine secretion screening identified molecules that induce levels of tumor necrosis factor alpha (TNF-α) and interleukin-8 (IL-8) comparable to the levels induced by phosphorylated hexa-acyl disaccharide (PHAD). The lead candidates demonstrated potent immunostimulation in mouse splenocytes, human primary blood mononuclear cells (PBMCs), and human monocyte-derived dendritic cells (DCs). This newly described system allows directed programming of lipid A synthesis and has the potential to generate a diverse array of TLR4 agonist candidates. IMPORTANCE There is an urgent need to develop effective vaccines against infectious diseases that continue to be major causes of morbidity and mortality worldwide. Making effective vaccines requires selecting an adjuvant to strengthen an appropriate and protective immune response. This work describes a practical method, bacterial enzymatic combinatorial chemistry (BECC), for generating functionally diverse molecules for adjuvant use. These molecules were analyzed in cell culture for their ability to initiate immune stimulatory activity. Several of the assays described herein show promising in vitro cytokine production and costimulatory molecule expression results, suggesting that the BECC molecules may be useful in future vaccine preparations.Kelsey A. GreggErin HarbertsFrancesca M. GardnerMark R. PelletierCorinne CayatteLi YuMichael P. McCarthyJason D. MarshallRobert K. ErnstAmerican Society for MicrobiologyarticleTLR4adjuvantsimmunomodulationinnate immunitylipid AlipopolysaccharideMicrobiologyQR1-502ENmBio, Vol 8, Iss 3 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	TLR4 adjuvants immunomodulation innate immunity lipid A lipopolysaccharide Microbiology QR1-502
spellingShingle	TLR4 adjuvants immunomodulation innate immunity lipid A lipopolysaccharide Microbiology QR1-502 Kelsey A. Gregg Erin Harberts Francesca M. Gardner Mark R. Pelletier Corinne Cayatte Li Yu Michael P. McCarthy Jason D. Marshall Robert K. Ernst Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
description	ABSTRACT Adjuvant properties of bacterial cell wall components like MPLA (monophosphoryl lipid A) are well described and have gained FDA approval for use in vaccines such as Cervarix. MPLA is the product of chemically modified lipooligosaccharide (LOS), altered to diminish toxic proinflammatory effects while retaining adequate immunogenicity. Despite the virtually unlimited number of potential sources among bacterial strains, the number of useable compounds within this promising class of adjuvants are few. We have developed bacterial enzymatic combinatorial chemistry (BECC) as a method to generate rationally designed, functionally diverse lipid A. BECC removes endogenous or introduces exogenous lipid A-modifying enzymes to bacteria, effectively reprogramming the lipid A biosynthetic pathway. In this study, BECC is applied within an avirulent strain of Yersinia pestis to develop structurally distinct LOS molecules that elicit differential Toll-like receptor 4 (TLR4) activation. Using reporter cell lines that measure NF-κB activation, BECC-derived molecules were screened for the ability to induce a lower proinflammatory response than Escherichia coli LOS. Their structures exhibit varied, dose-dependent, TLR4-driven NF-κB activation with both human and mouse TLR4 complexes. Additional cytokine secretion screening identified molecules that induce levels of tumor necrosis factor alpha (TNF-α) and interleukin-8 (IL-8) comparable to the levels induced by phosphorylated hexa-acyl disaccharide (PHAD). The lead candidates demonstrated potent immunostimulation in mouse splenocytes, human primary blood mononuclear cells (PBMCs), and human monocyte-derived dendritic cells (DCs). This newly described system allows directed programming of lipid A synthesis and has the potential to generate a diverse array of TLR4 agonist candidates. IMPORTANCE There is an urgent need to develop effective vaccines against infectious diseases that continue to be major causes of morbidity and mortality worldwide. Making effective vaccines requires selecting an adjuvant to strengthen an appropriate and protective immune response. This work describes a practical method, bacterial enzymatic combinatorial chemistry (BECC), for generating functionally diverse molecules for adjuvant use. These molecules were analyzed in cell culture for their ability to initiate immune stimulatory activity. Several of the assays described herein show promising in vitro cytokine production and costimulatory molecule expression results, suggesting that the BECC molecules may be useful in future vaccine preparations.
format	article
author	Kelsey A. Gregg Erin Harberts Francesca M. Gardner Mark R. Pelletier Corinne Cayatte Li Yu Michael P. McCarthy Jason D. Marshall Robert K. Ernst
author_facet	Kelsey A. Gregg Erin Harberts Francesca M. Gardner Mark R. Pelletier Corinne Cayatte Li Yu Michael P. McCarthy Jason D. Marshall Robert K. Ernst
author_sort	Kelsey A. Gregg
title	Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
title_short	Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
title_full	Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
title_fullStr	Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
title_full_unstemmed	Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery
title_sort	rationally designed tlr4 ligands for vaccine adjuvant discovery
publisher	American Society for Microbiology
publishDate	2017
url	https://doaj.org/article/9e94f7b9ae6c480f9062857e69a6fc14
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Rationally Designed TLR4 Ligands for Vaccine Adjuvant Discovery

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