Oropharyngeal microbiome evaluation highlights Neisseria abundance in active celiac patients

Iaffaldano, Laura; Granata, Ilaria; Pagliuca, Chiara; Esposito, Maria Valeria; Casaburi, Giorgio; Salerno, Giuliana; Colicchio, Roberta; Piccirillo, Marina; Ciacci, Carolina; Blanco, Giovanna Del Vecchio; Guarracino, Mario Rosario; Salvatore, Paola; Salvatore, Francesco; D’Argenio, Valeria; Sacchetti, Lucia

doi:10.1038/s41598-018-29443-1

Cited by 41 publications

(46 citation statements)

References 41 publications

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“…The genera Stomatobaculum and Oribacterium, belonging to the Lachnospiraceae family, are both obligately anaerobic bacteria in the human oral cavity and were significantly reduced in our obese group compared to controls. This result is suggestive of a continuum of microbiome composition between mouth and duodenum, previously observed by our group in a different disease model [39]. In agreement, decreased Lachnospiraceae were previously described in oral microbiome from obese compared to non-obese type 2 diabetic individuals [40].…”

Section: Discussionsupporting

confidence: 91%

Characterization of the Duodenal Mucosal Microbiome in Obese Adult Subjects by 16S rRNA Sequencing

et al. 2020

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The gut microbiota may have an impact on obesity. To date, the majority of studies in obese patients reported microbiota composition in stool samples. The aim of this study was to investigate the duodenal mucosa dysbiosis in adult obese individuals from Campania, a region in Italy with a very high percentage of obese people, to highlight microbial taxa likely associated with obesity. Duodenum biopsies were taken during upper gastrointestinal endoscopy in 19 obese (OB) and 16 lean control subjects (CO) and microbiome studied by 16S rRNA gene sequencing. Duodenal microbiome in our groups consisted of six phyla: Proteobacteria, Firmicutes, Actinobacteria, Fusobacteria, Bacteroidetes and Acidobacteria. Proteobacteria (51.1% vs. 40.1%) and Firmicutes (33.6% vs. 44.9%) were significantly (p < 0.05) more and less abundant in OB compared with CO, respectively. Oribacterium asaccharolyticum, Atopobium parvulum and Fusobacterium nucleatum were reduced (p < 0.01) and Pseudomonadales were increased (p < 0.05) in OB compared with CO. Receiver operating characteristic curve analysis showed Atopobium and Oribacterium genera able to discriminate with accuracy (power = 75% and 78%, respectively) OB from CO. In conclusion, increased Proteobacteria and decreased Firmicutes (Lachnospiraceae) characterized the duodenal microbiome of obese subjects. These data direct to further studies to evaluate the functional role of the dysbiotic-obese-associated signature.

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Section: Discussionsupporting

confidence: 91%

Characterization of the Duodenal Mucosal Microbiome in Obese Adult Subjects by 16S rRNA Sequencing

et al. 2020

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“…In conclusion, the N. flavescens strain that we identified in duodenum and oropharyngeal samples from patients with active CD (D'Argenio et al, 2016;Iaffaldano et al, 2018) To prepare the probiotic supernatant, we cultivated L. paracasei-CBA L74 in DMEM supplemented with 10% fetal calf serum (FCS, GIBCO), and 1-mM glutamine (GIBCO) until 10 9 CFU/ml as previously described (Sarno et al, 2014). The bacterial culture was then centrifuged at 3,000 rpm for 10 min and the supernatant was filtered through a 0-to 22-micron filter.…”

Section: Flavescensmentioning

confidence: 66%

“…Using 16S rRNA analysis of duodenal and oropharyngeal samples from CD patients and control subjects (Ctr), we previously identified a peculiar Neisseria flavescens strain in adults affected by CD (D'Argenio et al, 2016;Iaffaldano et al, 2018). This bacterial strain, isolated from the above samples, induced an immune-inflammatory response in human and murine dendritic cells, in CaCo-2 cells, and in ex vivo duodenal mucosal explants of Ctr subjects, thereby suggesting that it could play a role in CD (D'Argenio et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

“…3 | DISCUSSIONThe aims of this study were to investigate (a) if, parallel to the inflammatory response, the N. flavescens strain, which is increased in duodenal and oropharyngeal samples from active celiac patients(D'Argenio et al, 2016;Iaffaldano et al, 2018), induces changes in the CaCo-2 bioenergetics thus concurring to exacerbate the epithelial intestinal cell malfunction; (b) if the intracellular trafficking of N. flavescens and the immunogenic P31-43 peptide could influence each other; and (c) if the pretreatment of CaCo-2 cells with the L. paracasei-CBA supernatant could modify the effect of CD-N. flavescens-associated dysbiosis on the cell, despite the presence of the P31-43 peptide. Mitochondrial oxidative phosphorylation was significantly reduced in CD-N. flavescens-infected CaCo-2 cells versus NT cells, which together with the significantly increased ECAR/OCR ratio, suggested a shift in CD-N. flavescens-infected CaCo-2 cell metabolism towards a more glycolytic phenotype.…”

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confidence: 99%

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Celiac disease‐associated Neisseria flavescens decreases mitochondrial respiration in CaCo‐2 epithelial cells: Impact of Lactobacillus paracasei CBA L74 on bacterial‐induced cellular imbalance

Labruna

Nanayakkara

Pagliuca

et al. 2019

Cellular Microbiology

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We previously identified a Neisseria flavescens strain in the duodenum of celiac disease (CD) patients that induced immune inflammation in ex vivo duodenal mucosal explants and in CaCo‐2 cells. We also found that vesicular trafficking was delayed after the CD‐immunogenic P31‐43 gliadin peptide‐entered CaCo‐2 cells and that Lactobacillus paracasei CBA L74 (L. paracasei‐CBA) supernatant reduced peptide entry. In this study, we evaluated if metabolism and trafficking was altered in CD‐N. flavescens‐infected CaCo‐2 cells and if any alteration could be mitigated by pretreating cells with L. paracasei‐CBA supernatant, despite the presence of P31‐43. We measured CaCo‐2 bioenergetics by an extracellular flux analyser, N. flavescens and P31‐43 intracellular trafficking by immunofluorescence, cellular stress by TBARS assay, and ATP by bioluminescence. We found that CD‐N. flavescens colocalised more than control N. flavescens with early endocytic vesicles and more escaped autophagy thereby surviving longer in infected cells. P31‐43 increased colocalisation of N. flavescens with early vesicles. Mitochondrial respiration was lower (P < .05) in CD‐N. flavescens‐infected cells versus not‐treated CaCo‐2 cells, whereas pretreatment with L. paracasei‐CBA reduced CD‐N. flavescens viability and improved cell bioenergetics and trafficking. In conclusion, CD‐N. flavescens induces metabolic imbalance in CaCo‐2 cells, and the L. paracasei‐CBA probiotic could be used to correct CD‐associated dysbiosis.

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“…Moreover, we found that the a-CD-associated N. flavescens affected mitochondrial respiration in CaCo-2 epithelial cells [5]. Next, we investigated the oropharyngeal microbiome in CD patients and controls to evaluate whether this niche shared microbial composition with the duodenum [6]. We found that Neisseria spp.…”

Section: Introductionmentioning

confidence: 95%

Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis

et al. 2019

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Coeliac disease (CD) is a multifactorial autoimmune disorder and gut dysbiosis contributes to its pathogenesis. We previously profiled by 16S rRNA sequencing duodenal and oropharyngeal microbiomes in active CD (a-CD), gluten-free diet (GFD) patients, and controls (CO) and found significantly higher levels of Neisseria spp., with pro-inflammatory activities, in a-CD patients than in the other two groups. In this study, we developed a fast and simple qPCR-based method to evaluate the abundance of the oral Neisseria spp. and the diagnostic performances of the test in CD diagnosis. The Neisseria spp. abundances detected by quantitative PCR (qPCR) were: CO = 0.14, GFD = 0.15, a-CD = 2.08, showing a similar trend to those previously measured by next generation sequencing (NGS). In particular, Neisseria spp. values obtained by both methods were significantly higher (p < 0.001) in a-CD than in the other two groups GFD and CO—the latter almost overlapping. We calculated by ROC curve analysis the threshold of 1.12 ng/μL of Neisseria spp. to discriminate between CO+GFD and a-CD patients with 100% and 96.7% of diagnostic sensitivity and specificity, respectively. In conclusion, our data, if confirmed in other cohorts, suggest the q-PCR evaluation of oral Neisseria spp. could be a fast and simple method to assess CD-associated dysbiosis for diagnostic purposes.

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Oropharyngeal microbiome evaluation highlights Neisseria abundance in active celiac patients

Cited by 41 publications

References 41 publications

Characterization of the Duodenal Mucosal Microbiome in Obese Adult Subjects by 16S rRNA Sequencing

Characterization of the Duodenal Mucosal Microbiome in Obese Adult Subjects by 16S rRNA Sequencing

Celiac disease‐associated Neisseria flavescens decreases mitochondrial respiration in CaCo‐2 epithelial cells: Impact of Lactobacillus paracasei CBA L74 on bacterial‐induced cellular imbalance

Setup of Quantitative PCR for Oral Neisseria spp. Evaluation in Celiac Disease Diagnosis

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