ht29-mtx cell culture  (Inserm Transfert)

Journal: NPJ biofilms and microbiomes

Article Title: Role of mucus-bacteria interactions in Enterotoxigenic Escherichia coli (ETEC) H10407 virulence and interplay with human microbiome.

doi: 10.1038/s41522-022-00344-6

Figure Lengend Snippet: Fig. 1 ETEC adhesion to the mucus compartment. Adhesion of the ETEC strain H10407 to the mucus compartment was analyzed by three different in vitro assays. A ETEC bacteria (initial concentration: 107 CFU.ml−1) adhered in plate assays to type II mucin-agar layer (orange dots) or agar without mucin used as a negative control (black dots) after one hour exposure. B ETEC adhesion to Caco-2/ HT29-MTX co-culture model (orange dots) or Caco-2 cells only (black dots) after infection at MOI 100 for 3 h. C ETEC bacteria (initial concentration: 107 CFU.ml−1) adhered to type III mucin-alginate beads (orange dots) or alginate without mucin used as a negative control (black dots), after one hour and half static gastro-ileal digestion procedure. Figures represent the results of three independent experiments (in B, all technical replicates are repre- sented). Means are indicated by black bars. p-values are provided by unpaired t test with Welch’s correction (*p < 0.05; ****p < 0.0001).

Article Snippet: Caco-2 and HT29-MTX cell culture assays Caco-2 cells were purchased (Cell lines service, Eppelheim, Germany) and HT29-MTX cells were originated from Thecla Lesuffleur97.

Techniques: In Vitro, Bacteria, Concentration Assay, Negative Control, Co-Culture Assay, Infection

Journal: NPJ biofilms and microbiomes

Article Title: Role of mucus-bacteria interactions in Enterotoxigenic Escherichia coli (ETEC) H10407 virulence and interplay with human microbiome.

doi: 10.1038/s41522-022-00344-6

Figure Lengend Snippet: Fig. 3 Dynamics of virulence gene expression in planktonic and adhered ETEC bacteria during gastrointestinal transit and interactions with intestinal cells. ETEC virulence gene expression was analyzed by RT-qPCR in the gastric (A) and ileal (B) effluents of the TIM-1 model inoculated with 1010 CFU and in cellular experiments (MOI 100) involving Caco-2 cells cultivated with or without HT29-MTX mucus-secreting cells (C). Gene expression was analyzed over-time in the TIM-1 on planktonic bacteria or bacteria adhered to mucin beads (A, B) or intestinal cells (C). Results were expressed and colored according to fold-change expression compared to ETEC gene expression in the glass of water used to inoculate the TIM-1 model (T0) (A, B) or planktonic bacteria upon Caco-2 cells (C). Assayed genes were estP (ST toxin), eltB (LT toxin), leoA (LT toxin output), tolC (ST toxin outpout), tia (adhesin), fimH (minor component of type I pilus), yghJ (mucinase), eatA (mucinase) and rpos (environmental stress response). Results that are significantly different from each other according to Tukey’s multi-comparison are grouped under different yellow letters (p < 0.05).

Article Snippet: Caco-2 and HT29-MTX cell culture assays Caco-2 cells were purchased (Cell lines service, Eppelheim, Germany) and HT29-MTX cells were originated from Thecla Lesuffleur97.

Techniques: Gene Expression, Bacteria, Quantitative RT-PCR, Expressing, Comparison

Journal: NPJ biofilms and microbiomes

Article Title: Role of mucus-bacteria interactions in Enterotoxigenic Escherichia coli (ETEC) H10407 virulence and interplay with human microbiome.

doi: 10.1038/s41522-022-00344-6

Figure Lengend Snippet: Fig. 4 ETEC induction of Interleukin-8 production by mucin secreting or non-secreting intestinal cells. Interleukin-8 (IL-8) extracellular secretion (A) and intracellular production (B) by ETEC- infected Caco-2 (black dots) or Caco-2/HT29-MTX (orange dots) cells were measured by an ELISA assay. Intestinal cells were infected for a 3-hour period with 107 CFU.mL−1 (MOI 100) with ETEC strain H10407. Control experiments were performed without the bacteria. Results are expressed as fold changes compared to non-infected Caco-2/HT29-MTX cells. The data represents the replicates of at least 3 independent experiments with their means (black line). Statistical differences provided by Tukey’s multiple comparisons test are indicated (****p < 0.0001).

Article Snippet: Caco-2 and HT29-MTX cell culture assays Caco-2 cells were purchased (Cell lines service, Eppelheim, Germany) and HT29-MTX cells were originated from Thecla Lesuffleur97.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Control, Bacteria