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Bifidobacterium in the treatment of inflammatory disease

a technology of bifidobacterium and inflammatory disease, which is applied in the field of strains of bifidobacterium to achieve the effect of attenuating inflammatory activity

Inactive Publication Date: 2002-01-17
ENTERPRISE IRELAND TRADING AS BIORES IRELAND GLASNEVIN +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0114] Bifidobacterium species exert inhibitory effects on other bacteria by excluding long term colonisation by invasive pathogens. Their antagonistic activity is due to the production of acetic and lactic acid though fermentation (Scardovi, V. (1986) Bifidibacterium in Bergey's Manual of systemic bacteriology, Vol. 2. Eds. Sheath, P. H., Main, N. S., Sharpe, M. and Holdt, J. G., Williams and Wilkins Publishers, Baltimore Md., p1418). Very few reports exist on the production of antimicrobial compounds other than acids (Anand, S. K. et al. Cult. Dairy Prods. 1985; J. 2, 21-23). Bacteriocins and other compounds may influence the survival of a bacterium in an ecological niche and allow them to effectively dominate fermenting ecosystems. Such a feature is a good trait for a probiotic strain.
[0122] Lactobacilius salivarius subsp. Salivarius UCC 118 and Bifidobacterium longum infantis UCC 35624 were isolated from the ileal-caecal region of an adult human as described in Example 1. In this example, spontaneous rifampicin and streptomycin resistant derivatives of the strains were generated by plating cells, previously grown overnight and subsequently washed in quarter strength Ringer's solution on MRS and TPY agar containing 50 .mu.g / ml rifampicin (Sigma) respectively and MRS containing 400 .mu.g / ml streptomycin (Sigma). Plates were incubated for 2 days at 37.degree. C. anaerobically. The resulting antibiotic resistant derivatives were determined to be otherwise phenotypically similar to the parent strain. This selectable trait enabled the strains to be readily enumerated following gut transit.
[0139] Mice consuming UCC 35624 alone appeared in good health and again weight loss when compared to the controls was considerably less (FIGS. 7 and 8). It can be concluded therefore that consumption of UCC 35624 either alone or in combination with UCC 118 alleviates the symptoms of inflammatory bowel disease.
[0150] Overnight cultures of Bifidobacterium longum infantis were centrifuged and the cell-free culture supernatant was examined for the presence of cytokine inhibitors. Cell free supernatants were incubated with human TNF.alpha. for 20 minutes at 37.degree. C. TNF.alpha. levels were quantified thereafter by ELISA. Following exposure to the Bifidobacteria supernatant, TNF.alpha. levels were significantly reduced (FIG. 13). Thus, Bifidobacterium longum infantis UCC35624 secretes a factor that antagonises TNF.alpha. activity. Production of this factor by Bifidobacterium longum infantis at the surface of the gastrointestinal tract, in vivo, would significantly restrict the host inflammatory response.
[0160] The immune system has a large repertoire of specificities expressed by B and T cells. Some of these specificities will be directed to self-components. Self-recognition is normally controlled by clonal deletion and inactivation of self-reactive lymphocytes. However, there is a constant background of autoimmunity with antibodies to many proteins being found in serum. A breakdown in the self-nonself recognition system results in autoimmunity. When autoimmune disease does occur, the resulting immune response damages the tissue bearing the offending antigen. Immune complex deposition, type II hypersensitivity and cell-mediated reactions are the most important mechanisms by which immunopathological damage occurs. Examples of autoimmune diseases include, but are not limited to, systemic lupus erytematosus, rheumatoid arthritis, insulin dependent diabetes mellitus, myasthenia gravis and pernicious anaemia. Bifidobacterium longum infantis and Lactobacillus salivarius subsp. salivarius are immunomodulatory bacteria. Thus, consumption either as single components or in combination of these bacteria by patients suffering from autoimmune disease may restrict organ damage and help restore normal body homeostasis.
[0164] The introduction of probiotic organisms is accomplished by the ingestion of the microorganism in a suitable carrier. It would be advantageous to provide a medium that would promote the growth of these probiotic strains in the large bowel. The addition of one or more oligosaccharides, polysaccharides, or other prebiotics enhances the growth of lactic acid bacteria in the gastrointestinal tract (Gibson, G R. Br. J. Nutr. 1998;80 (4):S209-12). Prebiotics refers to any non-viable food component that is specifically fermented in the colon by indigenous bacteria thought to be of positive value, e.g. bifidobacteria, lactobacilli. Types of prebiotics may include those which contain fructose, xylose, soya, galactose, glucose and mannose. The combined administration of a probiotic strain with one or more prebiotic compounds may enhance the growth of the administered probiotic in vivo resulting in a more pronounced health benefit, and is termed synbiotic.

Problems solved by technology

While a number of probiotic bacteria of human origin are now being exploited commercially (e.g., L. acidophilus LA-1), many consumers, consumer organisations, and members of the scientific community are sceptical of such products and their publicised probiotic claims.

Method used

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  • Bifidobacterium in the treatment of inflammatory disease
  • Bifidobacterium in the treatment of inflammatory disease
  • Bifidobacterium in the treatment of inflammatory disease

Examples

Experimental program
Comparison scheme
Effect test

example 2

[0098] Acid Resistance

[0099] The first line of host defence that a microorganism reaches following human consumption is gastric acid in the stomach. A key factor influencing bacteria is survival in gastric juice. The survival and growth of Bifidobacterium longum infantis strains 35612 and 35624 in a low pH environment were examined The strains were routinely cultured in trypticase-peptone-yeast extract (TPY) medium at 37.degree. C. under strict anaerobic conditions (BBL Gas jars using the Merck Anaerocult A gas pak system) for 12-24 h. Human gastric juice was obtained from healthy subjects by aspiration through a nasogastric tube (Mercy Hospital, Cork, Ireland). It was immediately centrigued at 13,000 g for 30 min. to remove all solid particles, sterilised through 0.45 .mu.m filters and 0.2.mu.m filters and stored at 4.degree. C. The pH and pepsin activity were measured prior to experimental use. Pepsin activity was measured using the quantitative haemoglobin assay (Guantam, S. and ...

example 3

Bile Resistance

[0105] In the evaluation of the effectiveness of using lactic acid bacteria as beneficial members of the gastrointestinal tract, it is considered that resistance to bile acids is an important biological strain characteristic required for survival in this hostile environment and in addition they must not impinge on the health of the host by producing toxic compounds such as deoxycholic (DCA) and lithocholic acid (LCA) which have been implicated in a number of cytotoxic phenomena.

[0106] A number of Bifidobacterium longum infantis strains were streaked onto TPY agar plates supplemented with porcine bile (B-8631, Sigma Chemical Co. ltd., Poole) at concentrations of 0.3, 0.5, 1.0, 1.5, 5.0 and 7.5% (w / v) (Legrand-Defretin, R. et al., Lipids 1991; 26 (8), 578-583). Porcine bile is the closest in composition to human bile with respect to bile salt / cholesterol and phospholipid / cholesterol ratios. Plates were incubated at 37.degree. C. under anaerobic conditions and growth was...

example 4

Antimicrobial Activity

[0114] Bifidobacterium species exert inhibitory effects on other bacteria by excluding long term colonisation by invasive pathogens. Their antagonistic activity is due to the production of acetic and lactic acid though fermentation (Scardovi, V. (1986) Bifidibacterium in Bergey's Manual of systemic bacteriology, Vol. 2. Eds. Sheath, P. H., Main, N. S., Sharpe, M. and Holdt, J. G., Williams and Wilkins Publishers, Baltimore Md., p1418). Very few reports exist on the production of antimicrobial compounds other than acids (Anand, S. K. et al. Cult. Dairy Prods. 1985; J. 2, 21-23). Bacteriocins and other compounds may influence the survival of a bacterium in an ecological niche and allow them to effectively dominate fermenting ecosystems. Such a feature is a good trait for a probiotic strain.

[0115] The inhibitory spectra of various bifidobacterial strains was determined by the method of Tagg et al. CTagg. J. R. et al. Bacteriol. Rev. 1976; 40, 722-756). Cell free s...

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Abstract

A strain of Bifidobacterium isolated from resected and washed human gastrointestinal tract is significantly immunomodulatory following oral consumption in humans. The strain is useful in the prophylaxis and / or treatment of undesirable inflammatroy activity, especially gastrointestinal inflammatory activity such as inflammatory bowel disease or irritable bowel syndrome. The inflammatory activity may also be due to cancer.

Description

[0001] This invention relates to probiotic Bifidobacterium strains which have various applications in foodstuffs and in medicine. More particularly, the invention relates to probiotic strains of bifidobacteria which are capable of beneficially modifying and consequently alleviating observable symptoms in inflammatory disease.[0002] Consumers are becoming increasingly aware of matters which may be necessary for maintenance of their environment, health and nutrition. In response, scientific research has focussed upon the roles that diet, stress, and modem medical practices (e.g. antibiotics and radiotherapy) may play in threatening human health. In particular, population dynamics shifting towards older societies are increasing the incidence of illnesses which may be caused by deficient or compromised microflora such as gastrointestinal tract (GIT) infections, constipation, irritable bowel syndrome (IBS), inflammatory bowel disease (EBD)--Crohn's disease and ulcerative colitis, food al...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A23L1/30A23C9/123A61K9/14A61K9/20A61K9/48A61K39/00A61K45/00A61K47/02A61K47/04A61K47/06A61K47/08A61K47/18A61K47/26A61K47/42A61K47/44A61K47/46A61P1/00A61P1/04A61P1/12A61P19/02A61P29/00A61P31/04A61P31/12A61P35/00A61P37/04A61P37/06A61P43/00C12N1/20C12R1/01C12R1/225
CPCA23C9/1234A23L1/3014A23V2002/00A23Y2220/77A61K39/09A61K2039/52A61K2039/542C12N1/20C12R1/01A23V2200/3204A23V2200/324Y10S435/853Y10S435/822A23L33/135A61P1/00A61P1/04A61P1/12A61P19/02A61P19/04A61P29/00A61P31/04A61P31/12A61P31/14A61P35/00A61P37/00A61P37/02A61P37/04A61P37/06A61P43/00C12R2001/01C12N1/205A23V2400/179
Inventor COLLINS, JOHN KEVINO'SULLIVAN, GERALD CHRISTOPHERO'MAHONY, LIAMSHANAHAN, FERGUS
Owner ENTERPRISE IRELAND TRADING AS BIORES IRELAND GLASNEVIN
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