INTESTINAL INFECTION

Lactobacillus acidophilus (strain LB) from the resident adult human gastrointestinal microflora exerts activity against brush border damage promoted by a diarrhoeagenic Escherichia coli in human enterocyte-like cells

V Liévin-Le Moal, R Amsellem, A L Servin, M-H Coconnier

Institut National de la Santé et de la Recherche Médicale, Unité 510, Pathogénes et Fonctions des Cellules Epithéliales Polarisées, Faculté de Pharmacie, Université Paris XI, F-92296, Chãtenay-Malabry, France

Correspondence to:
Correspondence to:
A L Servin, Faculté de Pharmacie Paris XI, INSERM Unité 510, F-92296 Chãtenay-Malabry, France;
alain.servin{at}cep.u-psud.fr

Background and aims: The normal gastrointestinal microflora exerts a barrier effect against enteropathogens. The aim of this study was to examine whether lactobacilli, a minor genus of the resident gut microflora, exerts a protective effect against the cellular injuries promoted by the diarrhoeagenic Afa/Dr diffusely adhering Escherichia coli (Afa/Dr DAEC) C1845 strain in human intestinal cells.

Methods: Cultured human intestinal fully differentiated enterocyte-like Caco-2/TC7 cells were used. Antibacterial activity was examined by measuring the viability of the adhering C1845 bacteria. The distribution of brush border associated cytoskeleton and functional proteins was examined by immunofluorescence labelling coupled to confocal laser scanning microscopy analysis.

Results: The activity of Lactobacillus acidophilus strain LB isolated from the resident human gastrointestinal microflora was examined. A dose dependent decrease in viability of C1845 bacteria was observed after both direct contact in vitro between the spent culture supernatant (LB-SCS) and the bacteria, and when the bacteria were adherent on Caco-2/TC7 cells. Protection against the C1845 induced alterations in expression of F-actin, sucrase-isomaltase, dipeptidylpeptidase IV, alkaline phosphatase, and fructose transporter alterations was observed when the cells were exposed to LB-SCS.

Conclusion: L acidophilus strain isolated from the resident adult human gastrointestinal microflora, together with its antimicrobial activity, exerts a protective effect against the brush border lesions promoted by the diarrhoeagenic Afa/Dr DAEC strain C1845.

Keywords: intestinal cells; brush border lesions; gastrointestinal function; lactobacillus; antimicrobial; diarrhoea

Abbreviations: MV, microvillus; SI, sucrase-isomaltase; DPP IV, dipeptidylpeptidase IV; GPI, glycosylphosphatidylinositol; AP, alkaline phosphatase; TRITC, tetramethyl rhodamine isothiocyanate; FITC, fluorescein isothiocyanate; MRS broth, De Man, Rogosa, Sharpe broth; SCS, spent culture supernatant; TSA, tryptic soy agar; PBS, phosphate buffered saline; LDH, lactate dehydrogenase; cfu, colony forming units; CLSM, confocal laser scanning microscope; DMEM, Dulbecco's minimal essential medium

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