Holdemanella sp. bacterium and use thereof
A technology of bacteria, uses, applied in the field of HOLDEMANELLA SP. bacteria and uses, which can solve the problems of limited effectiveness, single treatment target, and no solution to obesity and the complexity of its complications
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Embodiment 1
[0093] Example 1. Isolation and identification of Holdemanella biformis bacteria.
[0094] Different gut bacteria were isolated from the feces of healthy volunteers. 1.25 g of feces were used and diluted in 10 mM phosphate buffered saline containing 0.05% cysteine (1:10 dilution) with a NaCl concentration of 130 mM (PBS) and processed on a Lab-Blender Stomacher 400 (Seward Medical, London, 35UK) for homogenization. Said dilution was inoculated in 37.5 ml of intestinal bacteria medium (IBM), the composition of which was based on the medium recommended in previous publications (Gibson, G.R., et al., Appl. Environ. Microbiol., 54(1 1) :2750-5, 1988; Lesmes, U et al., J.Agric.FoodChem., 56:5415-5421, 2008), some modifications devised by the inventors:
[0095] - main ingredient : Distilled water (1600ml), peptone water (4g), NaHCO 3 (4 grams), CaCl 2 (0.02g), Pectin (4g), Xylan (4g), Wheat Bran Extract (4g), Arabinogalactan (2g), Arabic Gum (2g), Starch (10g), Cheese Egg ...
Embodiment 2
[0107] Example 2. Selection of H. biformis based on its ability to regulate inflammation in vitro.
[0108] An in vitro assay was performed to comparatively assess the immunomodulatory properties of a collection of bacterial isolates, allowing selection of bacteria capable of inducing the greatest anti-inflammatory response in classical monocytes and, therefore, potentially therapeutic for the treatment of obesity-associated inflammation and alterations in glucose metabolism interest. To this end, cell suspensions of different bacteria were used as stimulators of the number of peripheral blood mononuclear cells (PBMC) and classical monocytes as well as the levels of the anti-inflammatory cytokine IL-4 relative to the pro-inflammatory cytokine IFNγ by flow cytometry technical measurement.
[0109] -Cultivation and stimulation of PBMC
[0110] From whole blood of healthy volunteers, peripheral blood mononuclear cells (PBMC) were isolated using a Ficoll gradient (Ficoll Paque...
Embodiment 3
[0116] Example 3. Characterization of the role of H.biformis in regulating energy and glucose metabolism in an obese animal model
[0117] Development and sampling of animal models of obesity
[0118] Adult male C57BL / 6 mice (6-8 weeks, Charles River, Les Oncins, France) were maintained and fed under controlled temperature (23°C), relative humidity (40-50%) and a 12-hour light / dark cycle A high-calorie diet (HFHSD; D12451, Research Diet, Brogaarden, Denmark) rich in fat (45% Kcal) and sucrose (17% Kcal) or a control diet with a standard fat content and no sucrose (CD, 10% from fat % Kcal; D12450K, study diet, Brogaarden, Denmark) for 14 weeks. Every day, mice fed a HFHSD diet received subjects (1×10 8 colony forming unit [CFU]) oral dose. Vehicle or placebo (10% skim milk) was administered to obese phenotype (HFHSD) control group and lean phenotype (CD) control group (n=10 mice per group) in the same manner. After 14 weeks, mice were sacrificed by cervical dislocation, a...
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