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A299 IDENTIFICATION OF PATHOGENIC BACTERIAL STRAINS IN PAEDIATRIC PATIENTS WITH INFLAMMATORY BOWEL DISEASES USING IMMUNOGLOBULIN G AS A MARKER OF VIRULENCE

Alipour, M ; Armstrong, H ; Valcheva, R S ; Zaidi, D ; Jovel, J ; Lou, Y ; Mason, A ; Wong, G ; Madsen, K ; Dieleman, L A ; Carroll, M W ; Huynh, H Q ; Wine, E

Journal of the Canadian Association of Gastroenterology, 2018-03, Vol.1 (suppl_1), p.519-520 [Periódico revisado por pares]

US: Oxford University Press

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  • Título:
    A299 IDENTIFICATION OF PATHOGENIC BACTERIAL STRAINS IN PAEDIATRIC PATIENTS WITH INFLAMMATORY BOWEL DISEASES USING IMMUNOGLOBULIN G AS A MARKER OF VIRULENCE
  • Autor: Alipour, M ; Armstrong, H ; Valcheva, R S ; Zaidi, D ; Jovel, J ; Lou, Y ; Mason, A ; Wong, G ; Madsen, K ; Dieleman, L A ; Carroll, M W ; Huynh, H Q ; Wine, E
  • Assuntos: Poster Presentations
  • É parte de: Journal of the Canadian Association of Gastroenterology, 2018-03, Vol.1 (suppl_1), p.519-520
  • Descrição: Abstract Background Inflammatory bowel diseases (IBD), including Crohn disease (CD) and ulcerative colitis (UC), are a group of chronic and severely debilitating gastrointestinal disorders, with an exacerbated immune response to the gut microbiota. Aims We hypothesized that pathogenic bacteria are more likely to be bound by immunoglobulin (Ig) G antibodies and identification of these strains could help identify mechanisms of immune activation. Methods Aspirate washes were obtained from the terminal ileum of pediatric IBD and non-IBD patients during endoscopy. Samples were fixed in paraformaldehyde and stringently washed to separate bacteria. Prior to fluorescence-activated cell sorting (FACS), samples were stained with propidium iodide (PI) and an anti-IgG fluorescent antibody with proper controls to differentiate bacteria bound by IgG (IgG+) from all others (IgG-). Validation in a select set of samples involved image cytometry. DNA was extracted from sorted bacteria using bead beating extraction and phenol/chloroform purification. Analysis of bacterial DNA samples was completed using the Ilumina MiSeq platform for 16S rRNA gene sequencing. Results Quality of patient bacterial DNA samples was used to identify those suitable to be sequenced resulting in 36 total washes from children without IBD (n=10), and with CD (n=17) or UC (n=9). Firmicutes and Bacteroidetes phyla were most commonly identified; however, no significant differences were found between IBD and non-IBD controls upon examining the composition of the ileal mucosa-associated microbiome. There was a 2 and 1.5 fold increase in the overall ratio of IgG+/IgG- bacteria in CD and UC, respectively. In CD, there was an increase in IgG+/IgG- ratio of both Bacteroidetes and Proteobacteria phyla. In UC this ratio was increased for Actinobacteria. IgG binding favored specific family level strains including Porphyromonadaceae (Bacteroidetes) and Enterobacteriaceae (Proteobacteria) in CD; Barnesiellaceae (Bacteroidetes) and Bifidobacteriaceae (Actinobacteria) in UC; and Clostridiaceae and Veillonellaceae (Firmicutes) in non-IBD, although considerable variation was noted between patients. Conclusions This study demonstrated selective increase of mucosa associated bacteria bound by IgG in the ileum of pediatric IBD patients compared to non-IBD. Further use of this method in larger cohort studies can identify individual microbes as therapeutic targets facilitating the development of targeted diagnostic and improved therapeutic approaches for IBD. Funding Agencies CCCCCFC, AIHS
  • Editor: US: Oxford University Press
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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