Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody

ABSTRACT Nontypeable Haemophilus influenzae (NTHi) is an important pathogen in individuals of all ages. The lipooligosaccharide (LOS) of NTHi has evolved a complex structure that can be attributed to a multiplicity of glycosyltransferases, the random switching of glycosyltransferase gene expression...

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Autores principales:	Michael A. Apicella, Jeremy Coffin, Margaret Ketterer, Deborah M. B. Post, Christopher J. Day, Freda E.-C. Jen, Michael P. Jennings
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2018
Materias:	biofilm ELISA lipooligosaccharide bactericidal activity keto-deoxyoctanoate nontypeable Haemophilus influenzae Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7c26c32dcf884786aa91e406c2cafa61
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spelling	oai:doaj.org-article:7c26c32dcf884786aa91e406c2cafa612021-11-15T16:00:15ZNontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody10.1128/mBio.01401-182150-7511https://doaj.org/article/7c26c32dcf884786aa91e406c2cafa612018-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.01401-18https://doaj.org/toc/2150-7511ABSTRACT Nontypeable Haemophilus influenzae (NTHi) is an important pathogen in individuals of all ages. The lipooligosaccharide (LOS) of NTHi has evolved a complex structure that can be attributed to a multiplicity of glycosyltransferases, the random switching of glycosyltransferase gene expression via phase variation, and the complex structure of its core region with multiple glycoform branch points. This article adds to that complexity by describing a multifunctional enzyme (LsgB) which optimally functions when the species is grown on a solid surface and which can add either a ketodeoxyoctanoate (KDO) or an N-acetylneuramic acid (Neu5Ac) moiety to a terminal N-acetyllactosamine structure of LOS. Our studies show that expression of lsgB is reduced four- to sixfold when NTHi is grown in broth. The substrate that the enzyme utilizes is dependent upon the concentration of free Neu5Ac (between 1 and 10 µg/ml) in the environment. In environments in which Neu5Ac is below that level, the enzyme utilizes endogenous CMP-KDO as the substrate. Our studies show that during in vivo growth in an NTHi biofilm, the KDO moiety is expressed by the organism. Monoclonal antibody 6E4, which binds KDO, is bactericidal for NTHi strains that express the KDO epitope at high levels. In a survey of 33 NTHi strains isolated from healthy and diseased individuals, the antibody was bactericidal (>90% kill) for 12 strains (36%). These studies open up the possibility of using a KDO-based glycoconjugate vaccine as part of a multicomponent vaccine against NTHi. IMPORTANCE Nontypeable Haemophilus influenzae is an important pathogen in middle ear infections in children, sinusitis in adults, and acute bronchitis in individuals with chronic obstructive lung disease. The organism is very well adapted to the human host environment, and this has hindered successful development of an effective vaccine. In this article, we describe a mechanism by which the bacteria decorates its surface lipooligosaccharide with a sugar unique to Gram-negative bacteria, ketodeoxyoctanoate (KDO). This sugar decoration is present during active infection and we have shown that an antibody directed against this sugar can result in killing of the organism. These data demonstrate that the lipooligosaccharide ketodeoxyoctanoate epitope may be a novel NTHi-specific candidate vaccine antigen.Michael A. ApicellaJeremy CoffinMargaret KettererDeborah M. B. PostChristopher J. DayFreda E.-C. JenMichael P. JenningsAmerican Society for MicrobiologyarticlebiofilmELISAlipooligosaccharidebactericidal activityketo-deoxyoctanoatenontypeable Haemophilus influenzaeMicrobiologyQR1-502ENmBio, Vol 9, Iss 4 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	biofilm ELISA lipooligosaccharide bactericidal activity keto-deoxyoctanoate nontypeable Haemophilus influenzae Microbiology QR1-502
spellingShingle	biofilm ELISA lipooligosaccharide bactericidal activity keto-deoxyoctanoate nontypeable Haemophilus influenzae Microbiology QR1-502 Michael A. Apicella Jeremy Coffin Margaret Ketterer Deborah M. B. Post Christopher J. Day Freda E.-C. Jen Michael P. Jennings Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody
description	ABSTRACT Nontypeable Haemophilus influenzae (NTHi) is an important pathogen in individuals of all ages. The lipooligosaccharide (LOS) of NTHi has evolved a complex structure that can be attributed to a multiplicity of glycosyltransferases, the random switching of glycosyltransferase gene expression via phase variation, and the complex structure of its core region with multiple glycoform branch points. This article adds to that complexity by describing a multifunctional enzyme (LsgB) which optimally functions when the species is grown on a solid surface and which can add either a ketodeoxyoctanoate (KDO) or an N-acetylneuramic acid (Neu5Ac) moiety to a terminal N-acetyllactosamine structure of LOS. Our studies show that expression of lsgB is reduced four- to sixfold when NTHi is grown in broth. The substrate that the enzyme utilizes is dependent upon the concentration of free Neu5Ac (between 1 and 10 µg/ml) in the environment. In environments in which Neu5Ac is below that level, the enzyme utilizes endogenous CMP-KDO as the substrate. Our studies show that during in vivo growth in an NTHi biofilm, the KDO moiety is expressed by the organism. Monoclonal antibody 6E4, which binds KDO, is bactericidal for NTHi strains that express the KDO epitope at high levels. In a survey of 33 NTHi strains isolated from healthy and diseased individuals, the antibody was bactericidal (>90% kill) for 12 strains (36%). These studies open up the possibility of using a KDO-based glycoconjugate vaccine as part of a multicomponent vaccine against NTHi. IMPORTANCE Nontypeable Haemophilus influenzae is an important pathogen in middle ear infections in children, sinusitis in adults, and acute bronchitis in individuals with chronic obstructive lung disease. The organism is very well adapted to the human host environment, and this has hindered successful development of an effective vaccine. In this article, we describe a mechanism by which the bacteria decorates its surface lipooligosaccharide with a sugar unique to Gram-negative bacteria, ketodeoxyoctanoate (KDO). This sugar decoration is present during active infection and we have shown that an antibody directed against this sugar can result in killing of the organism. These data demonstrate that the lipooligosaccharide ketodeoxyoctanoate epitope may be a novel NTHi-specific candidate vaccine antigen.
format	article
author	Michael A. Apicella Jeremy Coffin Margaret Ketterer Deborah M. B. Post Christopher J. Day Freda E.-C. Jen Michael P. Jennings
author_facet	Michael A. Apicella Jeremy Coffin Margaret Ketterer Deborah M. B. Post Christopher J. Day Freda E.-C. Jen Michael P. Jennings
author_sort	Michael A. Apicella
title	Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody
title_short	Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody
title_full	Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody
title_fullStr	Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody
title_full_unstemmed	Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody
title_sort	nontypeable <named-content content-type="genus-species">haemophilus influenzae</named-content> lipooligosaccharide expresses a terminal ketodeoxyoctanoate <italic toggle="yes">in vivo</italic>, which can be used as a target for bactericidal antibody
publisher	American Society for Microbiology
publishDate	2018
url	https://doaj.org/article/7c26c32dcf884786aa91e406c2cafa61
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Nontypeable <named-content content-type="genus-species">Haemophilus influenzae</named-content> Lipooligosaccharide Expresses a Terminal Ketodeoxyoctanoate <italic toggle="yes">In Vivo</italic>, Which Can Be Used as a Target for Bactericidal Antibody

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