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Probiotic bifidobacterium strain

A technology of bifidobacteria and strains, applied in bacteria, antibacterial drugs, antifungal agents, etc., can solve problems such as host injury

Active Publication Date: 2013-02-27
PRECISIONBIOTICS GRP LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, these control mechanisms must be able to distinguish non-pathogenic adherent bacteria from invading pathogens that would cause significant harm to the host

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Example 1: Isolation of Bifidobacterium longum AH1714

[0074] Bifidobacterium longum strain AH1714 was isolated from biopsies of healthy human subjects.

[0075]The screen for probiotics used a large number of human gastrointestinal tissue sections obtained from colorectal cancer screening. The human gastrointestinal mucosal tissue was transferred to a test tube containing phosphate buffered saline (PBS) (supplemented with 0.05% cysteine ​​hydrochloride). Triton X-100 (0.05%) was added to release adherent microorganisms in tissue samples. Tissue samples were then incubated for 10 minutes. Samples were vortexed and then plated on selective agar (De Man, Rogosa and Sharpe (MRS) agar + vancomycin and Wilkins-Chalgren agar + mupirocin, respectively) of adherent lactic acid isolated from gastrointestinal tissue bacilli and bifidobacteria. Colonies were isolated from the plate and streaked three times to ensure the purity of the clones. Microscopic examination, Gram s...

Embodiment 2

[0104] Example 2: Congo Red Agar Screening

[0105] Phenotypic screening of EPS-expressing bacterial strains was performed using Congo red agar selection. Briefly, 10 ml of modified Rogosa broth medium (+0.05% cysteine) was aseptically inoculated with a new colony of the bacterial strain and grown anaerobically at 37°C until cloudy (about 16 to about 24 hours). Broth cultures were aseptically streaked onto Congo red agar plates and incubated anaerobically at 37°C for 48 hours. EPS is believed to be produced as a by-product of growth and / or metabolism in certain strains, and EPS prevents the uptake of the Congo red dye, resulting in a creamy / white colony morphology. Strains that produce less EPS readily take up the Congo red dye, resulting in a pink / red colony morphology. Strains that do not produce EPS are stained red and appear almost transparent against the red agar background.

[0106] refer to figure 2 , the colony morphology of Bifidobacterium longum strain AH1714 ...

Embodiment 3

[0107] Example 3: Bifidobacterium strain 1714 induces a significantly elevated IL-10:IL-12 ratio .

[0108]Peripheral blood mononuclear cells (PBMC) were isolated from healthy human peripheral blood using BD Vacutainer CPT tubes (BD catalog 362761 ) according to the manufacturer's instructions. Wash PBMCs and resuspend them in Dulbecco's Modified Eagle Medium-Glutamax TM (Glutamax (glutamine substitute) + pyruvate + 4.5 g / l glucose (Gibco cat. no. 10569-010) 10% fetal bovine serum (Sigma cat. no. F4135), and 1% penicillin / streptomycin (Sigma cat. no. P0781). PBMC in (2×10 5 cells / well) in a flat-bottomed 96-well plate, and 20 μL of bacterial suspension (concentration of 1×10 7 CFU / mL). PBMC and bacteria at 37 °C / 5% CO 2 Co-culture in the incubator for 48 hours. After 2 days of culture, the plates were centrifuged at 300 xg, and the supernatant was removed and stored frozen at -80°C until analysis. Interleukin-10 (IL-10) and Interleukin-12p70 (IL-12p70) levels in cultur...

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Abstract

Probiotic Bifidobacterium strain AH1714 is significantly immunomodulatory following oral consumption. The strain is useful as an immunomodulatory biotherapeutic agent.

Description

Background of the invention [0001] The present invention relates to bifidobacterium strains and their use as probiotics, in particular as immunoregulatory biotherapeutics. [0002] The defense mechanisms that protect the human gastrointestinal tract from colonization with enteric bacteria are highly complex and involve both immune and non-immune aspects (1). Innate defense mechanisms include low pH of the stomach, bile salts, motility, mucin layer and antimicrobial compounds such as lysozyme (2). The immunological mechanism involves specific lymphoid aggregates beneath M cells, called Peyers patches, which are distributed throughout the small intestine and colon (3). Luminal antigens present at these locations stimulate appropriate T and B cell subsets, establish cytokine networks and secrete antibodies into the gastrointestinal tract (4). In addition, antigens can be presented via epithelial cells to intraepithelial lymphocytes and underlying lamina propria immune cells (5)...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/74C12N1/20A61K35/745
CPCC12N1/20A61K35/745A23V2002/00A23L1/3014A23Y2300/55A23C9/1234A23G9/363C12R1/01A23L33/135A61P1/00A61P1/02A61P1/04A61P1/12A61P1/16A61P11/00A61P11/06A61P13/12A61P15/00A61P17/00A61P17/02A61P17/06A61P17/10A61P19/02A61P19/10A61P25/00A61P25/28A61P29/00A61P3/00A61P3/04A61P31/00A61P31/04A61P31/10A61P31/12A61P31/18A61P35/00A61P35/04A61P37/00A61P37/02A61P37/06A61P37/08A61P5/00A61P7/02A61P7/06A61P9/00A61P9/10A61P3/10C12R2001/01C12N1/205A23V2400/533Y02A50/30A23V2200/3204A23V2200/324
Inventor 利亚姆·欧马奥尼巴里·凯利约翰·弗朗西斯·克赖恩蒂莫西·迪南艾琳·弗朗西斯·墨菲
Owner PRECISIONBIOTICS GRP LTD
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