WO2019129807A1 - Production de serpine - Google Patents

Production de serpine Download PDF

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Publication number
WO2019129807A1
WO2019129807A1 PCT/EP2018/097021 EP2018097021W WO2019129807A1 WO 2019129807 A1 WO2019129807 A1 WO 2019129807A1 EP 2018097021 W EP2018097021 W EP 2018097021W WO 2019129807 A1 WO2019129807 A1 WO 2019129807A1
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Prior art keywords
gluten
composition
strain cncm
longum
longum strain
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PCT/EP2018/097021
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English (en)
Inventor
Stéphane DUBOUX
Annick Mercenier
Gabriela Bergonzelli Degonda
Muzi TANGYU
Elena Verdu De Bercik
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Societe Des Produits Nestle S.A.
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Publication of WO2019129807A1 publication Critical patent/WO2019129807A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/04—Preserving or maintaining viable microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales

Definitions

  • the present invention relates to bacteria expressing serpin, methods for increasing serpin production in bacteria, and uses thereof.
  • Gluten-related disorders comprise all diseases triggered by gluten. They include, amongst other pathophysiology, celiac disease and non-celiac gluten sensitivity. Currently, the incidence of a wide spectrum of gluten-related disorders is growing all around the world, especially for celiac disease and non-celiac gluten sensitivity. Both diseases are triggered by ingestion of gluten. Both innate and adaptive immunity are implicated in celiac disease while innate immunity is implicated in non-celiac gluten sensitivity.
  • a life-long gluten free diet is the gold standard treatment for celiac disease and non-celiac gluten sensitivity patients, although it may have some limitations on the extraintestinal manifestations of the disease (Sedghizadeh et al., 2002, Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 94(4), 474-478). It has been shown that following a strict gluten free diet is very difficult as low level cross-contaminations are difficult to avoid and may happen through the whole food production chain, from grains growth to manufacturing processing (Mitchison et al., 1991 , Gut, 32(3), 260-265). Furthermore, it has been described that up to 3 g of hidden gluten might be consumed daily under a strict gluten free diet (Aziz et al., 2014, The American journal of gastroenterology, 109(9), 1498).
  • Celiac disease is prevalent especially in the United States and Europe where around 1 % of subjects had positive antibody tests (Dube et al., 2005, Gastroenterology, 128(4), S57-S67). It is a complex disorder which arises from a complicated interaction among various immunologic, genetic, and environmental factors (Alaedini & Green, 2005). It is triggered by the digestion of wheat gluten and other related cereal proteins such as rye and barley proteins.
  • Symptoms linked with celiac disease are growth retardation, irritability and pubertal delay in children, and many gastrointestinal symptoms such as discomfort, diarrhoea, occult stool, steatorrhea and flatulence, (Dube et al., 2005; Sedghizadeh et al., 2002).
  • Non-celiac gluten sensitivity is an emerging condition. It is defined as a clinical entity induced by the ingestion of gluten leading to intestinal and/or extraintestinal symptoms which could be improved by removing the gluten-containing foodstuff from the diet (Lundin & Alaedini, 2012). In addition to gliadin (the main cytotoxic antigen of gluten), other proteins/peptides present in gluten and gluten-containing cereals (wheat, rye, barley, and their derivatives) may play a role in the development of symptoms. Non-celiac gluten sensitivity is the most common syndrome of gluten-related disorders with prevalence rates between 0.5-13 % in the general population (on average 5 %) (Catassi et al., 2013, Nutrients, 5(10), 3839-3853).
  • Serine protease inhibitors are a superfamily of proteins found in eukaryotes (Gettins, 2002, Chemical reviews, 102(12), 4751-4804) and prokaryotes (Kantyka et al., Biochimie, 92(1 1 ), 1644-1656).
  • Elafin is human serine protease inhibitor which shows potent inhibitory capacity against various forms of elastases and proteinase (Ying & Simon, 1993, Biochemistry, 32(7), 1866-1874). Elafin is expressed throughout the epithelium of the gastrointestinal tract and its expression and induction is decreased in patients with inflammatory bowel disease and celiac disease (Baranger, Zani, Labas, Dallet- Choisy, & Moreau, 201 1 ; Motta et al., 2012).
  • elafin has been identified as a substrate for the cross-linking activity of TG2 (Baranger et al., 201 1 , PloS one, 6(6), e20976; Motta et al., Science translational medicine, 4(158), 158ra144-158ra144).
  • TG2 transglutaminase 2
  • elafin moderately inhibits transglutaminase 2 (TG2) thus inhibiting the deamidation of the digestion-resistant 33-mer gliadin peptide, which is one of the potential triggers of the adaptive immune response in CD (McCarville et al. 2015, Current opinion in pharmacology, 25, 7-12).
  • GMO genetically modified microorganism
  • B. longum subsp longum NCC 2705 displayed similar antiprotease activity to those of human serpin (Ivanov et al 2006, Journal of Biological Chemistry, 281 (25), 17246- 17252).
  • B. longum NCC 2705 was deposited with the Institute Pasteur according to the Budapest Treaty on 29 th January 2001 receiving the deposit no. CNCM 1-2618.
  • the present inventors have surprisingly found that spray-drying preparations of B. longum strain CNCM 1-2618 increases the level of serpin expression.
  • B. longum strain CNCM 1-2618 in the form of a spray-dried powder is associated with increased levels of serpin.
  • the B. longum strain CNCM 1-2618 is cultured in a medium comprising the sugar selected from lactose, fructose and raffinose, prior to spray drying.
  • a medium comprising the sugar selected from lactose, fructose and raffinose, prior to spray drying.
  • B. longum strain CNCM I- 2618 in the form of a spray dried powder.
  • the process comprises growing B. longum strain CNCM 1-2618 in a culture medium, followed by spray drying B. longum strain CNCM 1-2618.
  • the process comprises growing B. longum strain CNCM 1-2618 in a culture medium comprising a sugar selected from lactose, fructose and raffinose, or combinations thereof.
  • a sugar selected from lactose, fructose and raffinose, or combinations thereof.
  • lactose lactose
  • fructose fructose
  • raffinose a sugar selected from lactose, fructose and raffinose, or combinations thereof.
  • the culture medium comprises the sugar selected from lactose, fructose and raffinose at a concentration of 0.01 to 8 wt%, 0.02 to 6 wt%, 0.02 to 5 wt%, 0.02 to 4 wt%, 0.02 to 3 wt%, 0.02 to 2 wt %, 0.02 to 1 wt%, 0.02 to 0.5 wt %, 0.05 to 0.15 wt %, 0.08 to 0.12 wt %, or about 0.1 wt%.
  • B. longum strain CNCM I- 2618 in the form of a spray dried powder, wherein the spray dried powder is produced by growing B. longum strain CNCM 1-2618 in a culture medium comprising a sugar selected from lactose, fructose and raffinose, followed by spray drying the B. longum strain CNCM 1-2618.
  • composition comprising the B. longum strain CNCM 1-2618 in the form of a spray dried powder of the invention.
  • the composition may be, for example, a food, a medical food, a tube feed, or a nutritional supplement.
  • the food is selected from milk, yoghurt, curd, cheese, fermented milks, milk based fermented products, rice based products, milk based powders, infant formulae and pet food.
  • the composition is a pharmaceutical composition wherein the pharmaceutical composition comprises one or more pharmaceutically acceptable carriers, diluents and/or excipients.
  • the spray dried powder or composition of the invention for use in the treatment or prevention of conditions related to gluten sensitivity or involving the reduced activity of serine protease inhibitors.
  • the spray dried powder or composition of the invention for use in the treatment or prevention of a gluten-related disorder.
  • the spray dried powder or composition of the invention for use in the treatment or prevention of celiac disease, non-celiac gluten sensitivity, gluten ataxia, dermatitis herpetiformis and wheat allergy. According to another aspect of the present invention there is provided the spray dried powder or composition of the invention for use in the treatment or prevention of inflammatory bowel disease.
  • Figure 1 serpin mRNA fold induction in spray dried and freeze dried B. longum CNCM 1-2618. Fold induction calculate based on standard lab growth conditions (MRSc).“Frozen NCC 2705”: probiotic control, frozen biomass of CNCM 1-2618 (no drying applied).“SD NCC 2705”: B. longum CNCM 1-2618 produced by spray-drying.“FD NCC 2705”: B. longum CNCM 1-2618 produced by freeze-drying.
  • MRSc standard lab growth conditions
  • Figure 2 In vivo inhibition of elastase activity by spray dried and freeze dried B. longum CNCM 1-2618 preparations in NOD/DQ8 gluten sensitive mice. All mice were sensitized and challenged with gluten. Control: no administration of probiotic.
  • Frozen NCC 2705 probiotic control, frozen biomass of CNCM 1-2618, (no drying applied).
  • SD NCC 2705 B. longum CNCM 1-2618 produced by spray-drying.
  • FD NCC 2705 B. longum CNCM 1-2618 produced by freeze-drying.
  • composition of the present invention may be in the form of a food, a medical food, a tube feed, a nutritional composition, or a nutritional supplement.
  • nutritional supplement refers to a product which is intended to supplement the general diet of a subject.
  • the food is selected from milk, yogurt, curd, cheese, fermented milks, milk- based fermented products, rice based products, milk based powders, infant formulae and pet food.
  • the composition may be in the form of a medical food.
  • medical food refers to a food product specifically formulated for the dietary management of a medical disease or condition.
  • the medical food may be administered under medical supervision.
  • the medical food may be for oral ingestion or tube feeding.
  • the composition may be in the form of a tube feed.
  • tube feed refers to a product which is intended for introducing nutrients directly into the gastrointestinal tract of a subject by a feeding tube.
  • a tube feed may be administered by, for example, a feeding tube placed through the nose of a subject (such as nasogastric, nasoduodenal, and nasojejunal tubes), or a feeding tube placed directly into the abdomen of a subject (such as gastrostomy, gastrojejunostomy, or jejunostomy feeding tube).
  • composition may in the form of a pharmaceutical composition and may comprise one or more suitable pharmaceutically acceptable carriers, diluents and/or excipients.
  • suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like.
  • suitable diluents include ethanol, glycerol and water.
  • the choice of pharmaceutical carrier, excipient or diluent can be selected with regard to the intended route of administration and standard pharmaceutical practice.
  • the pharmaceutical compositions may comprise as, or in addition to, the carrier, excipient or diluent any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s) and/or solubilising agent(s).
  • suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flow lactose, beta-lactose, corn sweeteners, natural and synthetic gums, such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol.
  • Suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like.
  • Preservatives, stabilisers, dyes and even flavouring agents may be provided in the composition.
  • preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid.
  • Antioxidants and suspending agents may be also used.
  • Nutritionally acceptable carriers, diluents and excipients include those suitable for human or animal consumption and that are used as standard in the food industry. Typical nutritionally acceptable carriers, diluents and excipients will be familiar to the skilled person in the art.
  • composition may be in the form of a tablet, dragee, lozenge, capsule, gel cap, powder, granule, solution, emulsion, suspension, coated particle, spray-dried particle or pill.
  • composition may be in the form of a composition for topical administration, such as a gel, cream, ointment, emulsion, suspension or solution for topical administration.
  • a composition for topical administration such as a gel, cream, ointment, emulsion, suspension or solution for topical administration.
  • an ideal dose will depend on the subject to be treated, its health condition, sex, age, or weight, for example, and the route of administration.
  • the dose to be ideally used will consequently vary but can be determined easily by those of skill in the art.
  • the composition of the present invention comprises between 10 6 and 10 10 cfu and/or between 10 6 and 10 10 cells of B. longum strain CNCM 1-2618 per daily dose. It may also comprise between 10 6 and 10 11 cfu and/or between 10 6 and 10 11 cells of B. longum strain CNCM 1-2618 per g of the dry weight of the composition.
  • the B. longum strain CNCNCM 1-2618 may be cultured in a medium comprising lactose, fructose or raffinose, or mixtures thereof at a concentration of, for example, 0.02 to 0.50 wt %.
  • the B. longum strain CNCM 1-2618 may be cultured in a medium comprising lactose, fructose or raffinose, or mixtures thereof, at a concentration 0.05 to 0.15 wt %, 0.08 to 0.12 wt %, or about 0.10%.
  • Lactose is a disaccharide found in milk and composed of glucose and galactose.
  • Fructose is a monosaccharide found in many plants.
  • Raffinose is a trisaccharide also found in plants and composed of galactose, glucose and fructose.
  • the lactose, fructose, raffinose, or mixtures thereof may be added to a conventional culture medium comprising up to 8% of another sugar suitable to sustain B. longum growth, such as, but not limited to, glucose or sucrose.
  • Conventional culture medium suitable for growth of B. longum are well known to the person skilled in the art.
  • longum strain CNCM 1-2618 may be cultured in a medium comprising the lactose, fructose or raffinose, or mixtures thereof, at a concentration of 0.03 to 0.40, 0.04 to 0.30 or 0.05 to 0.20 wt %.
  • the culture medium may comprise the lactose, fructose or raffinose, or mixtures thereof, at a concentration of 0.03 to 0.15, 0.04 to 0.15, 0.05 to 0.15, 0.06 to 0.15 0.07 to 0.15, 0.08 to 0.15, 0.09 to 0.15 or 0.10 to 0.15 wt %.
  • the culture medium may comprise the lactose, fructose or raffinose, or mixtures thereof, at a concentration of 0.05 to 0.14, 0.05 to 0.13, 0.05 to 0.12 or 0.05 to 0.1 1 wt %.
  • the culture medium may comprise the lactose, fructose or raffinose, or mixtures thereof, at a concentration of 0.06 to 0.14, 0.07 to 0.13, 0.08 to 0.12, 0.09 to 0.1 1 or about 0.10 wt %.
  • lactose is used at the concentrations described above.
  • fructose is used at the concentrations described above.
  • raffinose is used at the concentrations described above.
  • the fermentation medium typically comprises
  • a nitrogen source such as yeast extract
  • a carbon source such as a sugar
  • various growth factors e.g minerals, vitamins etc.
  • MRS De Man, Rogosa and Sharpe
  • MRSc cysteine
  • the fermentation is preferably carried out in two steps, a starter fermentation being carried out prior to the main fermentation step.
  • the fermentation medium can be different for the starter and the main fermentation or may be identical.
  • the second step of the process is the concentration of the biomass. This can also be carried out using methods known to the person skilled in the art, such as for example centrifugation or filtration.
  • the total solid content of the biomass after concentration is preferably comprised between 10 and 35wt%, preferably between 14 and 35wt%, based on the total dry weight of the biomass (i.e. of the total amount of fermentation medium and produced microorganism).
  • the concentration may be preceded or combined with a washing step to remove residues of the fermentation medium and/or compounds produced during fermentation.
  • washing may be performed by concentrating biomass, re-suspending the concentrated biomass in a buffer, such as a phosphate buffer, or a similar composition and re-concentrating the biomass.
  • Spray drying is a conventional method of producing a dry powder by rapidly drying with a hot gas, well known in the art.
  • Suitable heated drying mediums for spray-draying include air and nitrogen.
  • the skilled person is able to identify suitable spray-drying conditions based on his knowledge. For example, the conditions described in EP0818529, which is entirely incorporated by reference, can be applied to spray-drying process.
  • the spray dried B. longum strain CNCM 1-2618 according to the present invention may be used in the treatment or prevention of conditions related to gluten sensitivity or involving reduced activity of serine protease inhibitors.
  • the spray dried powder or composition of the invention may be used in the treatment or prevention of inflammatory bowel disease, celiac disease, non-celiac gluten sensitivity, gluten ataxia, dermatitis herpetiformis and wheat allergy.
  • the condition is a gluten related disorder.
  • Gluten-related disorders encompass diseases triggered by gluten.
  • the terms“conditions related to gluten sensitivity” and“gluten- related disorders” are used interchangeably herein.
  • Gluten-related disorders include celiac disease, non-celiac gluten sensitivity, gluten ataxia, dermatitis herpetiformis and wheat allergy.
  • Celiac disease is one of the most common immune mediated disorders. It is a worldwide condition and is prevalent especially in the United States and Europe where around 1 % of subjects had positive antibody tests. Celiac disease is a complex disorder which arises from a complicated interaction among various immunologic, genetic, and environmental factors. It is triggered by the digestion of wheat gluten and other related cereal proteins such as rye and barley proteins. Symptoms linked with celiac disease are growth retardation, irritability and pubertal delay in children, and many gastrointestinal symptoms like discomfort, diarrhoea, occult stool, steatorrhea flatulence.
  • HLA-DQII human leukocyte antigens
  • HLA-DQ2.5/8 displaying those specific gluten peptides signals to helper T cells and other immune cells causing further damage in the small intestine.
  • Antibodies against gluten proteins and autoantibodies to connective tissue components (TG2) also associated with celiac disease progression (Alaedini & Green, 2005, Annals of internal medicine, 142(4), 289-298).
  • Non-celiac gluten sensitivity (also designated as non-celiac wheat sensitivity) is an emerging condition. It is defined as a clinical entity induced by the ingestion of gluten leading to intestinal and/or extra-intestinal symptoms which could be improved by removing the gluten-containing foodstuff from the diet (Lundin & Alaedini, 2012). The pathogenesis of non-celiac gluten sensitivity is not yet well understood. It has been shown that except for gliadin (main cytotoxic antigen of gluten), other proteins/peptides present in gluten and gluten-containing cereals (wheat, rye, barley, and their derivatives) may play a role in the development of symptoms.
  • gliadin main cytotoxic antigen of gluten
  • Non-celiac gluten sensitivity is the most common syndrome of gluten-related disorders with prevalence rates between 0.5-13 % in the general population (Catassi et al., 2013, Nutrients, 5(10), 3839-385). The diagnosis of non-celiac gluten sensitivity is made by exclusion of other gluten-related disorders. Dermatitis herpetiformis
  • Dermatitis herpetiformis is a chronic blistering skin autoimmune condition, characterized by the presence of skin lesions that have an extensive and symmetrical distribution, predominating in areas of greater friction, and affecting mainly both elbows, knees, buttocks, ankles, and may also affect the scalp and other parts of the body.
  • the lesions are vesicular-crusted and when flake off, they evolve to pigmented areas or a chromic an intense burning, itchy and blistering rash.
  • the age of onset is variable. It may start in childhood and adolescence but can also affect individuals of both sexes indistinctly at any age of their lives.
  • Gastrointestinal symptoms of wheat allergy are similar to those of celiac disease and non-celiac gluten sensitivity, but there is a different interval between exposure to wheat and onset of symptoms.
  • Wheat allergy has a fast onset (from minutes to hours) after the consumption of food containing wheat and can lead to anaphylaxis.
  • Gluten ataxia is a fast onset (from minutes to hours) after the consumption of food containing wheat and can lead to anaphylaxis.
  • Gluten ataxia is a gluten-related disorder. With gluten ataxia, damage takes place in the cerebellum, the balance center of the brain that controls coordination and complex movements like walking, speaking and swallowing. Gluten ataxia is the single most common cause of sporadic idiopathic ataxia. It accounts for 40% of ataxias of unknown origin and 15% of all ataxias.
  • Gluten ataxia is an immune-mediated disease triggered by the ingestion of gluten in genetically susceptible individuals. It should be considered in the differential diagnosis of all patients with idiopathic sporadic ataxia. The effectiveness of the treatment depends on the elapsed time from the onset of the ataxia until diagnosis. The death of neurons in the cerebellum as a result of gluten exposure of the subject is irreversible. Early diagnosis and treatment with a gluten free diet can improve ataxia and prevent its progression. Less than 10% of people with gluten ataxia present any gastrointestinal symptom, yet about 40% have intestinal damage. Sensitive markers of gluten ataxia include anti-gliadin antibodies. Immunoglobulin A (IgA) deposits against transglutaminase 2 (TG2) in the small bowel and at extraintestinal sites are proving to be additionally reliable.
  • IgA Immunoglobulin A deposits against transglutaminase 2 (TG2) in the small bowel and at extraintestinal sites are proving to be additionally reliable.
  • the spray dried Bifidobacterium longum strain CNCM 1-2618 or composition of the invention are preferably administered enterally.
  • Enteral administration may be oral, gastric, and/or rectal.
  • administration of the combination or composition described herein may, for example, be by an oral route or another route into the gastro-intestinal tract, for example the administration may be by tube feeding.
  • administration of the combination or composition described herein may be topical administration.
  • the subject may be a mammal such as a human, canine, feline, equine, caprine, bovine, ovine, porcine, cervine and primates.
  • the subject is a human.
  • the main fermentation was performed in a 10 L Biostar B Plus fermenter containing 5.4% of dextrose and 0.1 % of lactose. Live bacteria proportion and number were measured every hour by CytoFLEX to better adjust fermentation time. Bacteria were harvested after 14 h. Biomass was collected by centrifugation for 20 min at 5000 g. Protective agents (PA) (maltodextrin; sodium ascorbate; L-cysteine; L-alanine; L-lysine) were prepared in sterilized water and added in biomass with a ratio of 55 to 45 (dry mass ratio of PA/biomass) and stirred for homogenisation. Finally, half of it was subjected to spray drying and the other half to freeze drying. Total solid content (TS) was measured in biomass and biomass plus PA.
  • PA Protective agents
  • Spray drying was run in a Buchi system.
  • the set parameters were presented as follow: Inlet temperature 165 °C and outlet temperature 80-83 °C.
  • the system was preheated 20 min before.
  • Freeze-drying was performed in a Lyobeta 35 from Telstar. Freezing of the biomass was performed in liquid nitrogen. Primary drying was performed at 0.4 mbar, -15°C for 50 minutes. Secondary drying was performed at 20°C for 19h. Dried material was finally refined to obtain a fine powder.
  • Serpin mRNA detection and quantification Serpin mRNA levels were measured in the frozen biomass (frozen B. longum CNCM 1-2618, no drying process applied) and after drying.
  • RNA isolation 500 pi of bacterial cultures were suspended in 1 ml of RNA protect bacteria reagent (Qiagen, Germany) and briefly centrifuged 10 min at 5000 g to harvest cells for total RNA extraction.
  • RNA quality control and quantification - Quality and quantity control of RNA samples were analysed by electrophoresis using QIAxcel Advanced System (Qiagen, Germany).
  • RNA samples and RNA size marker were mixed separately with the same volume of RNA Denaturation Buffer and incubated 2 min at 70 °C, followed by 1 min cooling on ice. Then samples were diluted by QX RNA Dilution to 10 pi for analysing and measurement. Evaluation of RNA quality was performed using the ratio of 23S over 16S. RNA with a ratio value between 1 .6 and 2.3 were selected for later qRT-PCR.
  • PCR products were detected with SYBR green fluorescent dye and normalized with ROX reference dye.
  • the following primers were used for cDNA synthesis: for serpin gene: forward 5’- ACCAATCGCTGCTAAGTTCG-3’, reverse 5’-TCGCTGGCAAGAGAGTAGTC-3’; for Idh: forward, 5’-C G AAC GC C AT CT AC AT G CTC-3’ and reverse, 5’-AAGATCT G GTT CTCTTGCAG-3’ .
  • the primers for serpin were created based on B. longum NCC 2705 and DNA homology was checked in B. longum ATCC 15707. The reliability was verified by dissociation curve analysis.
  • mRNA fold expression analysis method The Pfaffl method was used to calculate relative transcription changes (Pfaffl, 2012) for which the equation is shown in Equation 1 : Equation 1
  • CFU Final Colony Forming Units after drying were equal to 1 .79E1 1 and 1.49E1 1 , respectively, for the freeze-dried and spray dried powder, representing a low viability loss upon drying of 0.2 log and 0.4 log, respectively.
  • B. longum serpins are able to inhibit elastase-like serine proteases through the same mechanism than that of eukaryotic serpins (Ivanov et al. , 2006; Journal of Biological Chemistry, 281 (25), 17246-17252.; Roberts et al., 2004; Journal of molecular evolution, 59(4), 437-447).
  • Ivanov et al. explored the inhibitory activity of B. longum CNCM 1-2618 (B. longum NCC 2705) serpin and found it could inhibit the activity of pancreatic elastase and neutrophil elastase.
  • B. longum CNCM 1-2618 B. longum NCC 2705
  • FD freeze- dried
  • SD spray-dried
  • Probiotics were administered daily during the challenge at a dosis of 1 E9 CFU per day. After the challenge, animals were sacrificed and intestinal content was collected for further elastase activity measurement.
  • the spray dried powder was surprisingly associated with greater elastase inhibition.
  • Such activity may confer advantageous properties on the bacteria such as increased protection in the gastrointestinal tract.

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Abstract

L'invention concerne l'utilisation du séchage par pulvérisation pour augmenter l'expression de serpine dans la souche Bifidobacterium Longum CNCM I-2618.
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021228805A1 (fr) * 2020-05-14 2021-11-18 Société des Produits Nestlé S.A. Procédés et compositions utilisant des bactéries produisant de la serpine
WO2021260163A1 (fr) * 2020-06-26 2021-12-30 Société des Produits Nestlé S.A. Production de serpine

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0818529A1 (fr) 1996-07-09 1998-01-14 Societe Des Produits Nestle S.A. Procédé de séchage par pulvérisation
EP2251020A1 (fr) * 2009-05-11 2010-11-17 Nestec S.A. Traitement haute température de courte durée qui génère des préparations microbiennes avec des profils anti-inflammatoires
WO2017001590A1 (fr) 2015-06-30 2017-01-05 Nestec S.A. Composition appropriée pour protéger des micro-organismes

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0818529A1 (fr) 1996-07-09 1998-01-14 Societe Des Produits Nestle S.A. Procédé de séchage par pulvérisation
EP2251020A1 (fr) * 2009-05-11 2010-11-17 Nestec S.A. Traitement haute température de courte durée qui génère des préparations microbiennes avec des profils anti-inflammatoires
WO2017001590A1 (fr) 2015-06-30 2017-01-05 Nestec S.A. Composition appropriée pour protéger des micro-organismes

Non-Patent Citations (41)

* Cited by examiner, † Cited by third party
Title
"Handbook of Pharmaceutical Excipients", 1994
"Remington's Pharmaceutical Sciences", 1985, MACK PUBLISHING CO.
ALAEDINI; GREEN, ANNALS OF INTERNAL MEDICINE, vol. 142, no. 4, 2005, pages 289 - 298
AUSUBEL, F.M. ET AL.: "Current Protocols in Molecular Biology", 1995, JOHN WILEY & SONS
AZIZ ET AL., THE AMERICAN JOURNAL OF GASTROENTEROLOGY, vol. 109, no. 9, 2014, pages 1498
BARANGER ET AL., PLOS ONE, vol. 6, no. 6, 2011, pages e20976
BUHNER S ET AL: "Protease signaling through protease activated receptor 1 mediate nerve activation by mucosal supernatants from irritable bowel syndrome but not from ulcerative colitis patients", PLOS ONE, vol. 13, no. 3, E0193943, 12 March 2018 (2018-03-12), pages 1 - 16, XP055474875, DOI: 10.1371/journal.pone.0193943 *
BURG; PILLINGER, CLINICAL IMMUNOLOGY, vol. 99, no. 1, 2001, pages 7 - 17
CATASSI ET AL., NUTRIENTS, vol. 5, no. 10, 2013, pages 3839 - 385
CATASSI ET AL., NUTRIENTS, vol. 5, no. 10, 2013, pages 3839 - 3853
DIETERICH ET AL., NATURE MEDICINE, vol. 3, no. 7, 1997, pages 797 - 801
DONG J. ET AL: "The Novel Role of a Serpin-Producing Probiotic in Gluten-Related Disorders", GASTROENTEROLOGY, 22 April 2017 (2017-04-22), XP085106565, ISSN: 0016-5085, DOI: 10.1016/S0016-5085(17)33412-1 *
DUBE ET AL., GASTROENTEROLOGY, vol. 128, no. 4, 2005, pages S57 - S67
GAIT, M.J.: "Oligonucleotide Synthesis: A Practical Approach", 1984, IRL PRESS
GALIPEAU ET AL., THE AMERICAN JOURNAL OF GASTROENTEROLOGY, vol. 109, no. 5, 2014, pages 748 - 756
GETTINS, CHEMICAL REVIEWS, vol. 102, no. 12, 2002, pages 4751 - 4804
HE XIANG ET AL: "[Proteomic analysis of Bifidobacteria longum strain NCC2705 grown on lactose and glucose]", WEISHENGWU XUEBAO - ACTA MICROBIOLOGICA SINICA, vol. 48, no. 11, 31 October 2008 (2008-10-31), pages 1451 - 1458, XP009505486, ISSN: 0001-6209 *
IVANOV D. ET AL: "A serpin from the gut bacterium Bifidobacterium longum inhibits eukaryotic elastase-like serine proteases", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 281, no. 25, 23 June 2006 (2006-06-23), pages 17246 - 17252, XP002586553, ISSN: 0021-9258, DOI: 10.1074/JBC.M601678200 *
IVANOV ET AL., JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 281, no. 25, 2006, pages 17246 - 17252
KANTYKA ET AL., BIOCHIMIE, vol. 92, no. 11, pages 1644 - 1656
KULLIN B. ET AL: "A functional analysis of the Bifidobacterium longum cscA and scrP genes in sucrose utilization", APPLIED MICROBIOLOGY AND BIOTECHNOLOGY, vol. 72, no. 5, 8 March 2006 (2006-03-08), pages 975 - 981, XP019441648, ISSN: 1432-0614, DOI: 10.1007/S00253-006-0358-X *
LIAN W.-C. ET AL: "SURVIVAL OF BIFIDOBACTERIA AFTER SPRAY-DRYING", INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY, vol. 74, no. 1-2, 25 March 2002 (2002-03-25), pages 79 - 86, XP009085030, ISSN: 0168-1605, DOI: 10.1016/S0168-1605(01)00733-4 *
LILLEY, D.M.; DAHLBERG, J.E.: "Methods in Enzymology: DNA Structures Part A: Synthesis and Physical Analysis of DNA", 1992, ACADEMIC PRESS
LIU D. ET AL: "Proteomics analysis of Bifidobacterium longum NCC2705 growing on glucose, fructose, mannose, xylose, ribose, and galactose", PROTEOMICS, vol. 11, no. 13, 1 July 2011 (2011-07-01), DE, pages 2628 - 2638, XP055475606, ISSN: 1615-9853, DOI: 10.1002/pmic.201100035 *
MCCARVILLE ET AL., APPL. ENVIRON. MICROBIOL., vol. 83, no. 19, 2017, pages e01323 - 17
MCCARVILLE ET AL., CURRENT OPINION IN PHARMACOLOGY, vol. 25, 2015, pages 7 - 12
MCCARVILLE J. L. ET AL: "A Commensal Bifidobacterium longum Strain Prevents Gluten-Related Immunopathology in Mice through Expression of a Serine Protease Inhibitor", APPLIED AND ENVIRONMENTAL MICROBIOLOGY, vol. 83, no. 19, E01323, 4 August 2017 (2017-08-04), US, pages 1 - 13, XP055474880, ISSN: 0099-2240, DOI: 10.1128/AEM.01323-17 *
MITCHISON ET AL., GUT, vol. 32, no. 3, 1991, pages 260 - 265
MOTTA ET AL., SCIENCE TRANSLATIONAL MEDICINE, vol. 4, no. 158, pages 158ra144 - 158ra144
PARCHE S. ET AL: "Lactose-over-Glucose Preference in Bifidobacterium longum NCC2705: glcP, Encoding a Glucose Transporter, Is Subject to Lactose Repression", JOURNAL OF BACTERIOLOGY, vol. 188, no. 4, 15 February 2006 (2006-02-15), pages 1260 - 1265, XP055065399, ISSN: 0021-9193, DOI: 10.1128/JB.188.4.1260-1265.2006 *
POLAK, J.M.; MCGEE, J.O'D: "In Situ Hybridization: Principles and Practice", 1990, OXFORD UNIVERSITY PRESS
REIMANN S. ET AL: "Development of a rapid screening protocol for selection of strains resistant to spray drying and storage in dry powder", BENEFICIAL MICROBES, vol. 1, no. 2, 1 June 2010 (2010-06-01), NL, pages 165 - 174, XP055474867, ISSN: 1876-2883, DOI: 10.3920/BM2009.0034 *
ROBERTS ET AL., JOURNAL OF MOLECULAR EVOLUTION, vol. 59, no. 4, 2004, pages 437 - 447
ROE, B.; CRABTREE, J.; KAHN, A.: "DNA Isolation and Sequencing: Essential Techniques", 1996, JOHN WILEY & SONS
SAMBROOK, J.; FRITSCH, E.F.; MANIATIS, T.: "Molecular Cloning: A Laboratory Manual", 1989, COLD SPRING HARBOR LABORATORY PRESS
SCHELL M. A. ET AL: "The genome sequence of Bifidobacterium longum reflects its adaptation to the human gastrointestinal tract", PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES, vol. 99, no. 22, 29 October 2002 (2002-10-29), pages 14422 - 14427, XP002313989, ISSN: 0027-8424, DOI: 10.1073/PNAS.212527599 *
SEDGHIZADEH ET AL., ORAL SURGERY, ORAL MEDICINE, ORAL PATHOLOGY, ORAL RADIOLOGY, AND ENDODONTOLOGY, vol. 94, no. 4, 2002, pages 474 - 478
SHIGA ET AL., BIOSCIENCE, BIOTECHNOLOGY, AND BIOCHEMISTRY, vol. 59, no. 12, 1995, pages 2348 - 2350
SOLLID, ANNUAL REVIEW OF IMMUNOLOGY, vol. 18, no. 1, 2000, pages 53 - 81
WEI X. ET AL: "Fructose Uptake in Bifidobacterium longum NCC2705 Is Mediated by an ATP-binding Cassette Transporter", JOURNAL OF BIOLOGICAL CHEMISTRY, vol. 287, no. 1, 18 November 2011 (2011-11-18), pages 357 - 367, XP055475596, ISSN: 0021-9258, DOI: 10.1074/jbc.M111.266213 *
YING; SIMON, BIOCHEMISTRY, vol. 32, no. 7, 1993, pages 1866 - 1874

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021228805A1 (fr) * 2020-05-14 2021-11-18 Société des Produits Nestlé S.A. Procédés et compositions utilisant des bactéries produisant de la serpine
WO2021260163A1 (fr) * 2020-06-26 2021-12-30 Société des Produits Nestlé S.A. Production de serpine

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