Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection

ABSTRACT The onset of Clostridium difficile infection (CDI) has been associated with treatment with wide-spectrum antibiotics. Antibiotic treatment alters the activity of gut commensals and may result in modified patterns of immune responses to pathogens. To study these mechanisms during CDI, we sep...

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Autores principales:	Mária Džunková, Andrés Moya, Jorge F. Vázquez-Castellanos, Alejandro Artacho, Xinhua Chen, Ciaran Kelly, Giuseppe D’Auria
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	16S rRNA gene sequencing Bayesian networks Clostridium difficile infection antibiotics dysbiosis fluorescence-activated cell sorting Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/be49b13898b14d98b11916d26d938a55
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spelling	oai:doaj.org-article:be49b13898b14d98b11916d26d938a552021-11-15T15:21:18ZActive and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection10.1128/mSphere.00101-162379-5042https://doaj.org/article/be49b13898b14d98b11916d26d938a552016-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00101-16https://doaj.org/toc/2379-5042ABSTRACT The onset of Clostridium difficile infection (CDI) has been associated with treatment with wide-spectrum antibiotics. Antibiotic treatment alters the activity of gut commensals and may result in modified patterns of immune responses to pathogens. To study these mechanisms during CDI, we separated bacteria with high cellular RNA content (the active bacteria) and their inactive counterparts by fluorescence-activated cell sorting (FACS) of the fecal bacterial suspension. The gut dysbiosis due to the antibiotic treatment may result in modification of immune recognition of intestinal bacteria. The immune recognition patterns were assessed by FACS of bacterial fractions either coated or not with intestinal secretory immunoglobulin A (SIgA). We described the taxonomic distributions of these four bacterial fractions (active versus inactive and SIgA coated versus non-SIgA coated) by massive 16S rRNA gene amplicon sequencing and quantified the proportion of C. difficile toxin genes in the samples. The overall gut microbiome composition was more robustly influenced by antibiotics than by the C. difficile toxins. Bayesian networks revealed that the C. difficile cluster was preferentially SIgA coated during CDI. In contrast, in the CDI-negative group Fusobacterium was the characteristic genus of the SIgA-opsonized fraction. Lactobacillales and Clostridium cluster IV were mostly inactive in CDI-positive patients. In conclusion, although the proportion of C. difficile in the gut is very low, it is able to initiate infection during the gut dysbiosis caused by environmental stress (antibiotic treatment) as a consequence of decreased activity of the protective bacteria. IMPORTANCE C. difficile is a major enteric pathogen with worldwide distribution. Its expansion is associated with broad-spectrum antibiotics which disturb the normal gut microbiome. In this study, the DNA sequencing of highly active bacteria and bacteria opsonized by intestinal secretory immunoglobulin A (SIgA) separated from the whole bacterial community by FACS elucidated how the gut dysbiosis promotes C. difficile infection (CDI). Bacterial groups with inhibitory effects on C. difficile growth, such as Lactobacillales, were mostly inactive in the CDI patients. C. difficile was typical for the bacterial fraction opsonized by SIgA in patients with CDI, while Fusobacterium was characteristic for the SIgA-opsonized fraction of the controls. The study demonstrates that sequencing of specific bacterial fractions provides additional information about dysbiotic processes in the gut. The detected patterns have been confirmed with the whole patient cohort independently of the taxonomic differences detected in the nonfractionated microbiomes.Mária DžunkováAndrés MoyaJorge F. Vázquez-CastellanosAlejandro ArtachoXinhua ChenCiaran KellyGiuseppe D’AuriaAmerican Society for Microbiologyarticle16S rRNA gene sequencingBayesian networksClostridium difficile infectionantibioticsdysbiosisfluorescence-activated cell sortingMicrobiologyQR1-502ENmSphere, Vol 1, Iss 3 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	16S rRNA gene sequencing Bayesian networks Clostridium difficile infection antibiotics dysbiosis fluorescence-activated cell sorting Microbiology QR1-502
spellingShingle	16S rRNA gene sequencing Bayesian networks Clostridium difficile infection antibiotics dysbiosis fluorescence-activated cell sorting Microbiology QR1-502 Mária Džunková Andrés Moya Jorge F. Vázquez-Castellanos Alejandro Artacho Xinhua Chen Ciaran Kelly Giuseppe D’Auria Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection
description	ABSTRACT The onset of Clostridium difficile infection (CDI) has been associated with treatment with wide-spectrum antibiotics. Antibiotic treatment alters the activity of gut commensals and may result in modified patterns of immune responses to pathogens. To study these mechanisms during CDI, we separated bacteria with high cellular RNA content (the active bacteria) and their inactive counterparts by fluorescence-activated cell sorting (FACS) of the fecal bacterial suspension. The gut dysbiosis due to the antibiotic treatment may result in modification of immune recognition of intestinal bacteria. The immune recognition patterns were assessed by FACS of bacterial fractions either coated or not with intestinal secretory immunoglobulin A (SIgA). We described the taxonomic distributions of these four bacterial fractions (active versus inactive and SIgA coated versus non-SIgA coated) by massive 16S rRNA gene amplicon sequencing and quantified the proportion of C. difficile toxin genes in the samples. The overall gut microbiome composition was more robustly influenced by antibiotics than by the C. difficile toxins. Bayesian networks revealed that the C. difficile cluster was preferentially SIgA coated during CDI. In contrast, in the CDI-negative group Fusobacterium was the characteristic genus of the SIgA-opsonized fraction. Lactobacillales and Clostridium cluster IV were mostly inactive in CDI-positive patients. In conclusion, although the proportion of C. difficile in the gut is very low, it is able to initiate infection during the gut dysbiosis caused by environmental stress (antibiotic treatment) as a consequence of decreased activity of the protective bacteria. IMPORTANCE C. difficile is a major enteric pathogen with worldwide distribution. Its expansion is associated with broad-spectrum antibiotics which disturb the normal gut microbiome. In this study, the DNA sequencing of highly active bacteria and bacteria opsonized by intestinal secretory immunoglobulin A (SIgA) separated from the whole bacterial community by FACS elucidated how the gut dysbiosis promotes C. difficile infection (CDI). Bacterial groups with inhibitory effects on C. difficile growth, such as Lactobacillales, were mostly inactive in the CDI patients. C. difficile was typical for the bacterial fraction opsonized by SIgA in patients with CDI, while Fusobacterium was characteristic for the SIgA-opsonized fraction of the controls. The study demonstrates that sequencing of specific bacterial fractions provides additional information about dysbiotic processes in the gut. The detected patterns have been confirmed with the whole patient cohort independently of the taxonomic differences detected in the nonfractionated microbiomes.
format	article
author	Mária Džunková Andrés Moya Jorge F. Vázquez-Castellanos Alejandro Artacho Xinhua Chen Ciaran Kelly Giuseppe D’Auria
author_facet	Mária Džunková Andrés Moya Jorge F. Vázquez-Castellanos Alejandro Artacho Xinhua Chen Ciaran Kelly Giuseppe D’Auria
author_sort	Mária Džunková
title	Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection
title_short	Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection
title_full	Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection
title_fullStr	Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection
title_full_unstemmed	Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection
title_sort	active and secretory iga-coated bacterial fractions elucidate dysbiosis in <named-content content-type="genus-species">clostridium difficile</named-content> infection
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/be49b13898b14d98b11916d26d938a55
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Active and Secretory IgA-Coated Bacterial Fractions Elucidate Dysbiosis in <named-content content-type="genus-species">Clostridium difficile</named-content> Infection

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