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Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes
Published on: May 8, 2016
Characterization of intestinal bacteria tightly bound to the human ileal epithelium
Saad Fakhry1, Nicola Manzo, Enrica D'Apuzzo
1Dipartimento di Biologia Strutturale e Funzionale, Università Federico II, via Cinthia 4, complesso MSA, 80126 Napoli, Italy.
Abstract:
In order to perform selective isolation of bacteria tightly bound to the human gut, ileal biopsies of healthy volunteers were treated to wash out the mucus layer and loosely bound bacterial cells. Rod-shaped anaerobic bacteria that had remained attached to the epithelial cells were isolated and identified at the species level. One isolate was identified as belonging to the Bifidobacterium breve species, while all the others were lactobacilli of only two species, Lactobacillus mucosae and Lactobacillus gasseri. Members of these species were found previously in intestinal samples, but their predominance among bacteria strictly associated with the epithelium was not suspected before and suggests that these species may represent a specific subpopulation of tissue-bound bacteria. Physiological analysis indicated that all isolates were able to produce antimicrobials, grow and form biofilm in simulated intestinal fluid after exposure to gastric conditions. Some isolates were able to degrade mucin while none showed cytotoxicity in vitro on HT29 cells. The tight association of the strains isolated with ileal epithelial cells is presumably indicative of a direct interaction with the host cells. For this reason and for the absence of cytotoxicity in vitro, those isolates can be proposed as potential probiotic strains for human use.
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