EP1229930A1 - Compositions and methods for treatment of allergic disorders - Google Patents

Compositions and methods for treatment of allergic disorders

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Publication number
EP1229930A1
EP1229930A1 EP00975685A EP00975685A EP1229930A1 EP 1229930 A1 EP1229930 A1 EP 1229930A1 EP 00975685 A EP00975685 A EP 00975685A EP 00975685 A EP00975685 A EP 00975685A EP 1229930 A1 EP1229930 A1 EP 1229930A1
Authority
EP
European Patent Office
Prior art keywords
bacteria
probiotic bacteria
administration
antigen
administered
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP00975685A
Other languages
German (de)
French (fr)
Other versions
EP1229930A4 (en
Inventor
Robert Llewellyn Clancy
Gerald Pang
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
DSM IP Assets BV
Original Assignee
Probiall Pty Ltd
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Filing date
Publication date
Application filed by Probiall Pty Ltd filed Critical Probiall Pty Ltd
Publication of EP1229930A1 publication Critical patent/EP1229930A1/en
Publication of EP1229930A4 publication Critical patent/EP1229930A4/en
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6869—Interleukin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/02—Bacterial antigens
    • A61K39/07—Bacillus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • 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
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/35—Allergens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/02—Nasal agents, e.g. decongestants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/06—Antiasthmatics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6863—Cytokines, i.e. immune system proteins modifying a biological response such as cell growth proliferation or differentiation, e.g. TNF, CNF, GM-CSF, lymphotoxin, MIF or their receptors
    • G01N33/6866—Interferon
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/335—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Lactobacillus (G)

Definitions

  • the invention relates to the field of allergic disease prevention and/or treatment, and in particular to probiotic bacteria which have the capacity to prevent and/or treat allergic disease.
  • BACKGROUND ART Allergy is a clinical syndrome affecting about one third of the population, manifest as rhinitis, asthma, eczema or food hypersensitivity.
  • target tissue changes play a role in determining the pattern of disease
  • the central abnormality (known as atopy) is the genetically influenced propensity to develop an IgE antibody response following antigen exposure.
  • atopy the central abnormality
  • the characteristics of the gut bacterial flora may drive IgE-promoting immunological mechanisms possibly by affecting the cytokine balance produced by CD4+ T lymphocytes.
  • the present invention provides a method of lowering IgE levels by administration of a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria-containing composition, to a subject in need thereof.
  • a method of prophylactic or therapeutic treatment of allergy by administration to a subject requiring such treatment a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria- containing composition preferably, the probiotic bacteria is, or the probiotic bacteria-containing composition includes. Lactobacillus.. Most preferably, the Lactobacillus is Lactobacill s acidophil s and/or Lactobacillus casei.
  • IgE is lowered from elevated levels induced by an allergen or as a consequence of an allergic disorder.
  • the probiotic bacteria, or a probiotic bacteria-containing composition is administered at the time of exposure to an allergen or shortly thereafter.
  • the term "exposure" with reference to an antigen or allergen includes natural exposure such as for example day-to-day contact with or ingestion of food products and the like, or seasonal exposure or re-exposure to allergens such as pollen or other air-borne allergens or by contact with skin and other body surfaces which may involve contact with synthetic materials or natural substances.
  • allergens such as pollen or other air-borne allergens or by contact with skin and other body surfaces which may involve contact with synthetic materials or natural substances.
  • the probiotic bacteria or compositions containing such bacteria are preferably administered at the time of exposure to the antigen/allergen or shortly thereafter.
  • the treatment is preferably commenced at the beginning of the season or shortly thereafter.
  • the term “exposure” includes natural exposure such as for example day-to-day contact with or ingestion of food products and the like, or seasonal exposure or re-exposure to allergens such as pollen or other air-borne allergens or by contact with skin and other body surfaces which may involve contact with synthetic materials or natural substances.
  • antigen/allergen when used with reference to antigen/allergen also includes artificial exposure or administration such as for example the injection of antigen/allergen in desensitisation procedures or under-tongue administration of antigen/allergen.
  • the antigen/allergen is preferably co-administered (co-presented) with the probiotic bacteria or a composition containing them.
  • probiotic bacteria may also be administered shortly after administration of the antigen/allergen.
  • the subject in need thereof is selected from the group consisting of high risk infants; those subjected to high risk occupational exposure to allergens; those exposed to high risk allergens; those having recognised allergy to specific allergens; and those prone to anaphylaxis.
  • the high risk infants are children of parents who both have allergic disease.
  • those subject to high risk occupation exposure to allergens are selected from the group consisting of aluminium smelter workers, woodworkers, chemical factory workers, and those working with latex-containing materials, especially gloves.
  • those exposed to high risk allergens are exposed to bee venom.
  • exposure to high risk allergens is parenteral exposure.
  • the specific allergens are present in the pre-pollen season, foods or latex-containing materials.
  • the trigger for the allergic response is insect envenomation, and food and drug sensitivities.
  • the probiotic bacteria or probiotic bacteria-containing composition is in tablet or capsule form.
  • the probiotic bacteria may be present in a food source such as a yoghurt or other dairy product.
  • the amount of probiotic bacteria administered to a human subject is at least 10 10 live bacteria. More preferably the amount administered is from about 10 10 to about 10 n live bacteria.
  • the required dosage amount will vary according to the severity of the allergic condition, the nature of the allergic condition, age of the subject and other standard clinical parameters. These parameters as well as the required dosage can be easily assessed by those skilled in the art. In human subjects it is preferred that the probiotic bacteria or a composition containing them be administered daily.
  • the present invention provides the use of live probiotic bacteria for the manufacture of a medicament for lowering IgE levels.
  • the present invention provides use of live probiotic bacteria for the manufacture of a medicament for treating allergy.
  • the present invention provides a method of identifying a bacterial species capable of lowering IgE levels in a mammal including: a) administration of the bacterial species to a mammal; b) administration of an allergen (antigen) to the mammal; and c) determination of the IgE antibody level in the mammal after treatment with the bacterial species and comparison with a control mammalian which a bacterial species
  • steps (a) and (b) were not administered, wherein steps (a) and (b) can be performed consecutively in any order or simultaneously.
  • the bacterial species is a Lactobacillus species and most preferably it is Lactobacillus acidophilus or Lactobacillus casei.
  • the animal model for identifying useful probiotic bacteria preferably makes use of the mouse.
  • other animal models may be developed on the same principle as disclosed herein.
  • the bacterial species is preferably administered orally however it may also be administered intraperitoneally and other means. Most preferably, the
  • 15 bacterial species is administered in an amount of 10 8 to 10" bacteria, and more preferably, in an amount of 0.6 to 1.0 x 10'° bacteria.
  • 1 to 20 oral doses are administered prior to administration of the antigen and most preferably 4 to 8 oral doses are administered prior to administration of the antigen.
  • administration of the oral doses is at 1 to 5 day intervals and, most preferably, at 2 day intervals.
  • 0 administration is over a 1 to 3 week period and, most preferably, over a 1 to 2 week period.
  • the antigen is administered at 5 a dose of 4 to 10 ⁇ g, and most preferably, at a dose of 8 ⁇ g.
  • the antigen is administered intraperitoneally.
  • the bacterial species is administered at the same time or 1 day after administration of the antigen/allergen.
  • the antigen/allergen can be administered after the bacterial species and in such circumstances the antigen/allergen is administered preferably 1 day after the 0 administration of 4 to 8 oral doses of bacteria.
  • administration of the bacteria species is continued after administration of the antigen. Most preferably, approximately 8 oral doses of bacteria are administered.
  • the bacteria are administered at 2 day intervals.
  • the IgE antibody is obtained from serum.
  • the serum is collected approximately 14 days after administration of the antigen.
  • the IgE antibody level is determined using an ELISA assay.
  • the skilled addressee will recognise that other IgE antibody assays may also be used.
  • the allergic disease is selected from the group consisting of asthma, eczema, hayfever and food allergy.
  • the present invention provides a bacterial species identified by the method of the fourth aspect.
  • the present invention provides a composition including a bacterial species according to the fourth aspect.
  • composition is in the form of a capsule or tablet or similar formulation however it may also be in the form of a food product.
  • a pharmaceutical composition including an effective amount of live bacterial species according to the fourth aspect, together with a pharmaceutically acceptable carrier, adjuvant, solvent or excipient.
  • the bacterial species is L. acidophilus.
  • a method of assessing efficacy of treatment with live probiotic bacteria or with a composition having live probiotic bacteria including the steps of: a measuring the level of salivary immunoglobulin subclass in a sample obtained before commencement of treatment, b measuring the level of salivary immunoglobulin subclass in a sample obtained after commencement of treatment c comparing the levels of salivary immunoglobulin subclass in a) and b), wherein the change in immunoglobulin subclass level is indicative of effective treatment.
  • the immunoglobulin subclass is IgGl or IgG2 and it will be understood that equivalent subclasses in various species can also be advantageously used.
  • a method of assessing efficacy of treatment with live probiotic bacteria or with a composition having live probiotic bacteria including the steps of: a measuring the level of LL-4 or IFN- ⁇ in a sample obtained before commencement of treatment, b measuring the level of IL-4 or IFN- ⁇ in a sample obtained after commencement of treatment c comparing the levels of IL-4 or IFN- ⁇ in a) and b), wherein the change in IL-4 or
  • IFN- ⁇ level is indicative of effective treatment. It will be clear however that any know cytokine marker for the Thl or the Th2 response will be suitable in place of IFN- ⁇ or IL-4.
  • Figure 1 Suppression of IgE antibody response in mice fed probiotic bacteria.
  • Figure 5 Suppression of IgE response to OVA, and cytokine and IgG antibody subclass response, by L acidophilus
  • a mouse animal model was developed which enabled identification of probiotic bacteria capable of downregulating the IgE response to an antigen/allergen.
  • the animal used is a mouse (for example C57BL/6).
  • the animals are fed the candidate bacterial species (alive, dose 10 8 -10" , frequency every 1 -5 days, for 1 -3 weeks).
  • the animals are fed the bacteria at the same time as the antigen was being introduced or shortly thereafter, for example the following day.
  • Mice were immunised intraperitoneally with antigen (for example ovalbumin) at an appropriate dose (eg 4-1 O ⁇ g, preferably 8 ⁇ g), with specific and total IgE measured in serum (eg. at 14 days after immunisation).
  • antigen for example ovalbumin
  • specific and total IgE measured in serum eg. at 14 days after immunisation.
  • feeding with test bacteria continues throughout the experiment (see examples).
  • comparisons are made with killed organisms and dose-response is estimated.
  • timing of administration of probiotic bacteria in relation to antigen/allergen challenge is examined. The examples demonstrate: (i) that frequent dosage with live probiotic bacteria at an appropriate does can reduce the IgE response to antigen/allergen.
  • oral administration of the bacterial species affects the systemic IgE antibody response (following injected antigen) and thus may be of value in subjects who have anaphylaxis (eg insect envenomation, and food and drug sensitivities).
  • probiotic bacteria can downregulate IgE production following antigen administration or exposure to antigen - thus continuous oral ingestion of appropriate doses of live bacteria selected by this method may be of value in the ongoing management of allergic disease.
  • administration of live probiotic bacteria is particularly effective
  • co-administration of probiotic bacteria with the antigen/allergen, or administration of probiotic bacteria shortly after the antigen/allergen challenge is particularly effective
  • the probiotic bacteria can be formulated into various compositions and preferably the compositions are pharmaceutical compositions in the form of capsules, tablets, powders and the like.
  • Such formulations can be prepared by known means, using pharmaceutically acceptable carriers, excipients, solvents or adjuvants. Such procedures and ingredients are well know and amply described in standard texts and manuals, for example "Remington: The Science and Practice of Pharmacy", 1995, Mack Publishing Co. Easton, PA 18042, USA, which is incorporated herein by reference.
  • the probiotic bacteria may also be formulated into food products by the usual well known means.
  • Bacteria ⁇ Lactobacillus acidophilus or Lactobacillus casei) were grown in MRS agar petri dishes (3.8% w/v, Oxoid, Basingstoke, UK). Plates were incubated in 5% CO 2 and air in a humid atmosphere at 37°C for 48 hours.
  • PBS phosphate buffered saline
  • mice Thirty female C57/B16 SPF mice (Animal Resource Centre, Perth, WA, Australia) were fed with 4 or 8 oral doses of 0.6-1.0 x 10 10 L acidophilus or L casei in 0.2 mL PBS or PBS alone given two days apart.
  • mice were injected intraperitoneally with 7 ⁇ g of egg albumin (ovalbumin. OVA, Sigma-Aldrich, St Loius, Missouri, USA) and 4 mg aluminium hydroxide (Amphojel, Whitehall Laboratories, Sydney, Australia) in 0.2 mL PBS or with PBS alone.
  • egg albumin ovalbumin. OVA, Sigma-Aldrich, St Loius, Missouri, USA
  • aluminium hydroxide aluminium hydroxide
  • mice Immunised and control mice were then placed on a feeding regime consisting of 8 oral doses of 0.6-1.0 x 10'° bacteria in PBS or PBS alone given two days apart. Mice were bled by the saphenous vein 2 days after the last dose and the serum collected for IgE antibody determination using an ELISA assay.
  • Example 2 Suppression of IgE response to OVA by L acidophilus after allergen sensitisation
  • mice were fed 10 10 L acidophilus before or 24 hrs after sensitisation with 8 ⁇ g ovalbumin (OVA) in alum per mouse.
  • Control mice were sham fed with normal saline.
  • a total of 10 feeds was administered before they were assessed for levels of OVA-specific IgE antibody and total IgE in serum.
  • L acidophilus was more effective if administered (or co-administered) at the time of exposure to OVA than when administered prior to allergen exposure, as shown by the suppression of OVA-specific IgE antibody and total IgE responses (Figure 2 ).
  • Data are mean + SEM from 5-10 mice. *, p ⁇ 0.05 compared with control values from saline-fed sensitised mice.
  • Example 3 Suppression of IgE response to OVA by L. acidophilus is dose- dependent Mice were fed orally with 10 8 . 10 9 or 10 10 L acidophilus before they were sensitised 24 hrs later with OVA in alum. Control mice were sham fed PBS. Each dose was administered 10 times every 2 days for 21 days. One week following the final dose, serum IgE and OVA-specific IgE antibody were measured. A dose-dependent suppression of IgE and OVA-specific IgE antibody was noted with a statistically significant effect with 10 l ⁇ bacteria ( Figure 3). Data are mean + SEM from 10 mice. *, p ⁇ 0.05 compared with control values from saline-fed sensitised mice.
  • mice were fed live or formalin-killed 10 10 L acidophilus and then sensitised with OVA as per standard protocol described in Example 3.
  • Figure 4 shows that live bacteria were more effective in suppressing IgE response than killed bacteria. Data are mean + SEM from 5 mice. *, p ⁇ 0.05 compared with values from saline-fed sensitised mice.
  • Example 5 Suppression of IgE response to OVA by L acidophilus correlates with cytokine and IgG antibody subclass response
  • mice fed 10'° L acidophilus produced higher amounts of IFN- ⁇ and lower amounts of IL-4 in the spleen, a finding consistent with the suppression of IgE response.
  • the contrasting cytokine patterns correlated with the production of salivary IgG subclass antibodies which showed an upregulation of IgG2a antibody and a downregulation of IgGl , respectively ( Figure 5 ).
  • Data are mean + SEM from 10 mice. *, p ⁇ 0.05; **, p ⁇ 0.01 compared with values from saline-fed sensitised mice.

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Abstract

The present invention relates to methods of lowering IgE levels and to methods of treating allergy, by administration of a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria-containing composition. The present invention also relates to methods for identifying useful probiotic bacteria and to compositions containing probiotic bacteria.

Description

"COMPOSITIONS AND METHODS FOR TREATMENT OF ALLERGIC
DISORDERS"
TECHNICAL FIELD
The invention relates to the field of allergic disease prevention and/or treatment, and in particular to probiotic bacteria which have the capacity to prevent and/or treat allergic disease.
BACKGROUND ART Allergy is a clinical syndrome affecting about one third of the population, manifest as rhinitis, asthma, eczema or food hypersensitivity. Although target tissue changes play a role in determining the pattern of disease, the central abnormality (known as atopy) is the genetically influenced propensity to develop an IgE antibody response following antigen exposure. Increasingly it is recognised that the characteristics of the gut bacterial flora may drive IgE-promoting immunological mechanisms possibly by affecting the cytokine balance produced by CD4+ T lymphocytes. Modification of the gut flora may modulate the balance of this T cell response, and indeed differences in faecal quantitative bacteriology have been described within populations of children who differ in their incidence of asthma. A means by which to achieve a reduction in the level of IgE is desirable.
The literature that exists describes the use of yoghurt which contains certain potentially beneficial bacteria but the content of such bacteria in yoghurt is relatively low (usually 105 to 107 organisms) and is largely unknown. Further, many studies show that commercially produced yoghurts contain large numbers of other, not necessarily beneficial bacteria, and often only dead bacteria. Such food products are consumed in an ad hoc and uncontrolled manner. There is no indication or suggestion of how such products, or how much, should be consumed to achieve beneficial effects.
It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.
SUMMARY OF THE INVENTION It has unexpectedly been found that certain bacterial species when introduced live into the digestive system in sufficient quantity have the capacity to downregulate IgE antibody, and thus prevent and/or treat allergic disease. Further, the timing of administration of the bacteria in relation to antigen/allergen exposure may also be of significance in certain circumstances.
According to a first aspect, the present invention provides a method of lowering IgE levels by administration of a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria-containing composition, to a subject in need thereof.
According to a second aspect there is provided a method of prophylactic or therapeutic treatment of allergy by administration to a subject requiring such treatment a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria- containing composition.. Preferably, the probiotic bacteria is, or the probiotic bacteria-containing composition includes. Lactobacillus.. Most preferably, the Lactobacillus is Lactobacill s acidophil s and/or Lactobacillus casei.
Preferably, IgE is lowered from elevated levels induced by an allergen or as a consequence of an allergic disorder. Preferably, the probiotic bacteria, or a probiotic bacteria-containing composition, is administered at the time of exposure to an allergen or shortly thereafter.
In the context of the present invention the term "exposure" with reference to an antigen or allergen includes natural exposure such as for example day-to-day contact with or ingestion of food products and the like, or seasonal exposure or re-exposure to allergens such as pollen or other air-borne allergens or by contact with skin and other body surfaces which may involve contact with synthetic materials or natural substances. In these circumstances the probiotic bacteria or compositions containing such bacteria are preferably administered at the time of exposure to the antigen/allergen or shortly thereafter. In seasonal exposure or re-exposure to allergens, the treatment is preferably commenced at the beginning of the season or shortly thereafter. However, the term
'"exposure" when used with reference to antigen/allergen also includes artificial exposure or administration such as for example the injection of antigen/allergen in desensitisation procedures or under-tongue administration of antigen/allergen. In these circumstances the antigen/allergen is preferably co-administered (co-presented) with the probiotic bacteria or a composition containing them. However, probiotic bacteria may also be administered shortly after administration of the antigen/allergen. Preferably, the subject in need thereof is selected from the group consisting of high risk infants; those subjected to high risk occupational exposure to allergens; those exposed to high risk allergens; those having recognised allergy to specific allergens; and those prone to anaphylaxis. Preferably, the high risk infants are children of parents who both have allergic disease. Preferably, those subject to high risk occupation exposure to allergens are selected from the group consisting of aluminium smelter workers, woodworkers, chemical factory workers, and those working with latex-containing materials, especially gloves. Preferably, those exposed to high risk allergens are exposed to bee venom. Preferably, exposure to high risk allergens is parenteral exposure. Preferably, in those having recognised allergic disease to specific allergens, the specific allergens are present in the pre-pollen season, foods or latex-containing materials. Preferably, in those prone to anaphylaxis, the trigger for the allergic response is insect envenomation, and food and drug sensitivities.
Preferably, the probiotic bacteria or probiotic bacteria-containing composition is in tablet or capsule form. However, it will be clear to those skilled in the art that the probiotic bacteria may be present in a food source such as a yoghurt or other dairy product.
Preferably the amount of probiotic bacteria administered to a human subject is at least 1010 live bacteria. More preferably the amount administered is from about 1010 to about 10 n live bacteria.
The required dosage amount will vary according to the severity of the allergic condition, the nature of the allergic condition, age of the subject and other standard clinical parameters. These parameters as well as the required dosage can be easily assessed by those skilled in the art. In human subjects it is preferred that the probiotic bacteria or a composition containing them be administered daily.
According to a third aspect the present invention provides the use of live probiotic bacteria for the manufacture of a medicament for lowering IgE levels.
According to a fourth aspect the present invention provides use of live probiotic bacteria for the manufacture of a medicament for treating allergy.
According to a fourth aspect, the present invention provides a method of identifying a bacterial species capable of lowering IgE levels in a mammal including: a) administration of the bacterial species to a mammal; b) administration of an allergen (antigen) to the mammal; and c) determination of the IgE antibody level in the mammal after treatment with the bacterial species and comparison with a control mammalian which a bacterial species
5 was not administered, wherein steps (a) and (b) can be performed consecutively in any order or simultaneously.
Preferably, the bacterial species is a Lactobacillus species and most preferably it is Lactobacillus acidophilus or Lactobacillus casei.
] o The animal model for identifying useful probiotic bacteria preferably makes use of the mouse. However other animal models may be developed on the same principle as disclosed herein.
In the animal model the bacterial species is preferably administered orally however it may also be administered intraperitoneally and other means. Most preferably, the
15 bacterial species is administered in an amount of 108 to 10" bacteria, and more preferably, in an amount of 0.6 to 1.0 x 10'° bacteria. Preferably, 1 to 20 oral doses are administered prior to administration of the antigen and most preferably 4 to 8 oral doses are administered prior to administration of the antigen. Preferably, administration of the oral doses is at 1 to 5 day intervals and, most preferably, at 2 day intervals. Preferably, 0 administration is over a 1 to 3 week period and, most preferably, over a 1 to 2 week period.
The preferred antigen/allergen used in the animal model is egg albumin (OVA) however it will be clear to those skilled in the art that other antigens can be used as allergens in this model should that be required. Preferably, the antigen is administered at 5 a dose of 4 to 10 μg, and most preferably, at a dose of 8 μg. Preferably, the antigen is administered intraperitoneally. Preferably, the bacterial species is administered at the same time or 1 day after administration of the antigen/allergen. However, in certain embodiments the antigen/allergen can be administered after the bacterial species and in such circumstances the antigen/allergen is administered preferably 1 day after the 0 administration of 4 to 8 oral doses of bacteria. Preferably, administration of the bacteria species is continued after administration of the antigen. Most preferably, approximately 8 oral doses of bacteria are administered. Preferably, the bacteria are administered at 2 day intervals.
Preferably, the IgE antibody is obtained from serum. Preferably, the serum is collected approximately 14 days after administration of the antigen. Most preferably, the IgE antibody level is determined using an ELISA assay. However, the skilled addressee will recognise that other IgE antibody assays may also be used.
Preferably, the allergic disease is selected from the group consisting of asthma, eczema, hayfever and food allergy. According to a fifth aspect, the present invention provides a bacterial species identified by the method of the fourth aspect.
According to a sixth aspect, the present invention provides a composition including a bacterial species according to the fourth aspect.
Preferably the composition is in the form of a capsule or tablet or similar formulation however it may also be in the form of a food product.
According to a seventh aspect there is provided a pharmaceutical composition including an effective amount of live bacterial species according to the fourth aspect, together with a pharmaceutically acceptable carrier, adjuvant, solvent or excipient.
Preferably the bacterial species is L. acidophilus. According to an eighth aspect there is provided a method of assessing efficacy of treatment with live probiotic bacteria or with a composition having live probiotic bacteria, including the steps of: a measuring the level of salivary immunoglobulin subclass in a sample obtained before commencement of treatment, b measuring the level of salivary immunoglobulin subclass in a sample obtained after commencement of treatment c comparing the levels of salivary immunoglobulin subclass in a) and b), wherein the change in immunoglobulin subclass level is indicative of effective treatment.
Preferably the immunoglobulin subclass is IgGl or IgG2 and it will be understood that equivalent subclasses in various species can also be advantageously used. According to a ninth aspect there is provided a method of assessing efficacy of treatment with live probiotic bacteria or with a composition having live probiotic bacteria, including the steps of: a measuring the level of LL-4 or IFN-γ in a sample obtained before commencement of treatment, b measuring the level of IL-4 or IFN-γ in a sample obtained after commencement of treatment c comparing the levels of IL-4 or IFN-γ in a) and b), wherein the change in IL-4 or
IFN-γ level is indicative of effective treatment. It will be clear however that any know cytokine marker for the Thl or the Th2 response will be suitable in place of IFN-γ or IL-4.
BRIEF DESCRIPTION OF DRAWINGS
Figure 1 : Suppression of IgE antibody response in mice fed probiotic bacteria.
Figure 2: Suppression of IgE response to OVA by L acidophilus before/after allergen sensitisation
Figure 3: Dose dependency of suppression of IgE response to OVA by L. acidophilus
Figure 4: Suppression of IgE response to OVA by live and killed L acidophilus
Figure 5: Suppression of IgE response to OVA, and cytokine and IgG antibody subclass response, by L acidophilus
DESCRIPTION OF THE PREFERRED EMBODIMENT A preferred embodiment of the invention will now be described by way of example only.
A mouse animal model was developed which enabled identification of probiotic bacteria capable of downregulating the IgE response to an antigen/allergen. Typically the animal used is a mouse (for example C57BL/6). The animals are fed the candidate bacterial species (alive, dose 108-10" , frequency every 1 -5 days, for 1 -3 weeks). In certain advantageous embodiments the animals are fed the bacteria at the same time as the antigen was being introduced or shortly thereafter, for example the following day. Mice were immunised intraperitoneally with antigen (for example ovalbumin) at an appropriate dose (eg 4-1 Oμg, preferably 8μg), with specific and total IgE measured in serum (eg. at 14 days after immunisation). In certain examples feeding with test bacteria continues throughout the experiment (see examples). In other examples comparisons are made with killed organisms and dose-response is estimated. In yet other examples the timing of administration of probiotic bacteria in relation to antigen/allergen challenge is examined. The examples demonstrate: (i) that frequent dosage with live probiotic bacteria at an appropriate does can reduce the IgE response to antigen/allergen.
(ii) that not all probiotic bacteria have the same capacity to reduce the IgE response - thus the model can be used to select candidate bacteria for human use in management or prevention of allergic disease. (iii) Lactobacillus acidophilus is particularly effective in blunting the IgE response to novel antigens.
(iv) that oral administration of the bacterial species affects the systemic IgE antibody response (following injected antigen) and thus may be of value in subjects who have anaphylaxis (eg insect envenomation, and food and drug sensitivities). (v) probiotic bacteria can downregulate IgE production following antigen administration or exposure to antigen - thus continuous oral ingestion of appropriate doses of live bacteria selected by this method may be of value in the ongoing management of allergic disease.
(vi) administration of live probiotic bacteria is particularly effective (vii) co-administration of probiotic bacteria with the antigen/allergen, or administration of probiotic bacteria shortly after the antigen/allergen challenge is particularly effective
(viii) to be effective doses of probiotic bacteria in excess of those found in conventional food sources are usually required The bacterial species identified by the method are useful
( 1 ) for the prevention of allergic disease
(a) in high risk infants (eg those with both parents with allergy)
(b) in those subjected to high risk occupation exposure (eg aluminium smelter workers, woodworkers, chemical factory workers and those working with latex-containing material, especially gloves)
(c) following parenteral exposure to high risk allergens, eg bee venom (d) in subjects having recognised allergy to specific allergens (eg pre-pollen season, foods, latex, etc)
(2) for the suppression of established allergic disease such as asthma, eczema, hayfever and food allergy. The probiotic bacteria can be formulated into various compositions and preferably the compositions are pharmaceutical compositions in the form of capsules, tablets, powders and the like. Such formulations can be prepared by known means, using pharmaceutically acceptable carriers, excipients, solvents or adjuvants. Such procedures and ingredients are well know and amply described in standard texts and manuals, for example "Remington: The Science and Practice of Pharmacy", 1995, Mack Publishing Co. Easton, PA 18042, USA, which is incorporated herein by reference.
The probiotic bacteria may also be formulated into food products by the usual well known means.
The invention will now be described more particularly with reference to non- limiting examples.
EXAMPLES Example 1: Suppression of IgE antibody response to OVA following oral administration of Lactobacillus.
Bacteria {Lactobacillus acidophilus or Lactobacillus casei) were grown in MRS agar petri dishes (3.8% w/v, Oxoid, Basingstoke, UK). Plates were incubated in 5% CO2 and air in a humid atmosphere at 37°C for 48 hours.
Bacteria were harvested, washed twice in phosphate buffered saline (PBS) and then resuspended to 3-5 x 10'° bacteria per mL in PBS.
Thirty female C57/B16 SPF mice (Animal Resource Centre, Perth, WA, Australia) were fed with 4 or 8 oral doses of 0.6-1.0 x 1010 L acidophilus or L casei in 0.2 mL PBS or PBS alone given two days apart.
One day after the last oral dose, mice were injected intraperitoneally with 7 μg of egg albumin (ovalbumin. OVA, Sigma-Aldrich, St Loius, Missouri, USA) and 4 mg aluminium hydroxide (Amphojel, Whitehall Laboratories, Sydney, Australia) in 0.2 mL PBS or with PBS alone.
Immunised and control mice were then placed on a feeding regime consisting of 8 oral doses of 0.6-1.0 x 10'° bacteria in PBS or PBS alone given two days apart. Mice were bled by the saphenous vein 2 days after the last dose and the serum collected for IgE antibody determination using an ELISA assay.
The results are shown in Figure 1. Suppression of IgE antibody response to OVA was observed in mice given oral doses of L acidophilus and L casei compared with immunised mice given PBS. This level of suppression was statistically significant when L acidophilus was used.
Example 2: Suppression of IgE response to OVA by L acidophilus after allergen sensitisation
C57/B16 mice were fed 1010 L acidophilus before or 24 hrs after sensitisation with 8 μg ovalbumin (OVA) in alum per mouse. Control mice were sham fed with normal saline. In each group, a total of 10 feeds was administered before they were assessed for levels of OVA-specific IgE antibody and total IgE in serum. L acidophilus was more effective if administered (or co-administered) at the time of exposure to OVA than when administered prior to allergen exposure, as shown by the suppression of OVA-specific IgE antibody and total IgE responses (Figure 2 ). Data are mean + SEM from 5-10 mice. *, p < 0.05 compared with control values from saline-fed sensitised mice. Baseline variation reflects different experiments and conditions ) Example 3: Suppression of IgE response to OVA by L. acidophilus is dose- dependent Mice were fed orally with 108. 109 or 1010 L acidophilus before they were sensitised 24 hrs later with OVA in alum. Control mice were sham fed PBS. Each dose was administered 10 times every 2 days for 21 days. One week following the final dose, serum IgE and OVA-specific IgE antibody were measured. A dose-dependent suppression of IgE and OVA-specific IgE antibody was noted with a statistically significant effect with 10lϋ bacteria (Figure 3). Data are mean + SEM from 10 mice. *, p < 0.05 compared with control values from saline-fed sensitised mice.
Extrapolation to human subjects indicates the requirement for a dose in the range of approximately 10'° tol O12 live L acidophilus each day to effectively downregulate IgE antibody-mediated allergic disease. This is considerably in excess of the numbers of live Lactobacilli which can be practically introduced via currently available food products (often about 105- 107 per serving). Example 4: Suppression of IgE response to OVA is dependent on live culture of L acidophilus
Mice were fed live or formalin-killed 1010 L acidophilus and then sensitised with OVA as per standard protocol described in Example 3. Figure 4 shows that live bacteria were more effective in suppressing IgE response than killed bacteria. Data are mean + SEM from 5 mice. *, p < 0.05 compared with values from saline-fed sensitised mice. Example 5: Suppression of IgE response to OVA by L acidophilus correlates with cytokine and IgG antibody subclass response
Mice fed 10'° L acidophilus produced higher amounts of IFN-γ and lower amounts of IL-4 in the spleen, a finding consistent with the suppression of IgE response. In addition, the contrasting cytokine patterns correlated with the production of salivary IgG subclass antibodies which showed an upregulation of IgG2a antibody and a downregulation of IgGl , respectively ( Figure 5 ). Data are mean + SEM from 10 mice. *, p < 0.05; **, p < 0.01 compared with values from saline-fed sensitised mice. These results indicate that IgG subclass antibody in saliva is an effective surrogate marker for monitoring response of allergic disease to intervention therapy. Thus measurement of equivalent subclasses in humans would be an appropriate laboratory test for effective desensitisation therapy eg. increase in IgG2 anti-allergen antibody and /or decrease in IgGl antibody in saliva (or other secretions) is indicative of a response to therapy.
Although the invention has been described with reference to specific examples, it will be appreciated by those skilled in the art that the invention may be embodied in many other forms. In particular, it will be clear to the skilled addressee that the methods and compositions described above can be applied to any probiotic which can lower IgE levels in a subject.

Claims

CLAIMS:
1. Method of lowering IgE levels by administration of a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria-containing composition, to a subject in need thereof.
2. Method of prophylactic or therapeutic treatment of allergy by administration to a subject requiring such treatment a therapeutically effective amount of live probiotic bacteria, or a live probiotic bacteria-containing composition..
3. Method according to claim 1 or claim 2, wherein the probiotic bacteria is, or the probiotic bacteria-containing composition includes, Lactobacillus.
4. Method according to any one of claims 1 to 3, wherein the Lactobacillus is
Lactobacillus acidophilus and/or Lactobacillus casei.
5. Method according to any one of claims 1 to 4, wherein the probiotic bacteria, or a probiotic bacteria-containing composition is administered at the time of exposure to an antigen/allergen or shortly thereafter.
6. Method according to claim 5. wherein the exposure is natural and/or seasonal exposure or re-exposure to antigen/allergen.
7. Method according to claim 5, wherein the exposure is artificial exposure to antigen/allergen.
8. Method according to claim 7. wherein the artificial exposure is the injection of antigen/allergen.
9. Method according to claim 7, wherein the artificial exposure is under-tongue administration of antigen/allergen.
10. Method according to any one of claims 1 to 9, wherein the amount of probiotic bacteria administered is at least 1010 live bacteria.
1 1. Method according to claim 10, wherein the amount of probiotic bacteria administered is from about 1010 to about 10'2 bacteria.
12. Method according to any one of claims 1 to 1 1, wherein the probiotic bacteria or the probiotic-bacteria containing composition is administered daily.
13. Method according to any one of claims 1 to 12, wherein the probiotic bacteria, or the probiotic bacteria-containing composition, is in a solid dosage form.
14. Method according to claim 13, wherein the probiotic bacteria, or the probiotic bacteria-containing composition, is in tablet or capsule form.
15. Method according to any one of claims 1 to 12, wherein the probiotic bacteria, or the probiotic bacteria-containing composition, is in the form of a food product.
16. Method according to any one of claims 1 to 15, wherein the subject has recognised allergic disease to specific allergens and wherein the specific allergens are present in the
5 pollen season, foods or latex-containing materials.
17. Method according to any one of claims 1 to 16, wherein the subject is prone to anaphylaxis and wherein the trigger for the allergic response is insect envenomation, food and/or drug sensitivities.
18. Method according to any one of claims 1 to 17, wherein the subject is exposed to 10 an allergen parenterally.
19. Use of live probiotic bacteria for the manufacture of a medicament for lowering IgE levels.
20. Use of live probiotic bacteria for the manufacture of a medicament for treating allergy.
15 21. Method of identifying a bacterial species capable of lowering IgE levels in a mammal including: a) administration of the bacterial species to a mammal; b) administration of an allergen (antigen) to the mammal; and c) determination of the IgE antibody and/or total IgE level in the mammal after 0 treatment with the bacterial species and comparison with a control mammal in which a bacterial species was not administered, wherein steps (a) and (b) can be performed consecutively in any order or simultaneously.
22. Method according to claim 21, wherein the bacterial species is a Lactobacillus 5 species.
23. Method according to claim 22, where the bacterial species is Lactobacillus acidophilus.
24. Method according to any one of claims 21 to 23, wherein 1 to 20 oral doses of bacteria are administered prior to administration of the antigen. 0
25. Method according to claim 24, wherein 4 to 8 oral doses of bacteria are administered.
26. Method according to any one of claims 21 to 25, wherein administration of the oral doses of bacteria is at 1 to 5 day intervals.
27. Method according to claim 26, wherein administration of the oral doses of bacteria is at 2 day intervals.
28. Method according to any one of claims 21 to 27, wherein the administration is over a 1 to 3 week period.
29. Method according to claim 28, wherein the administration is over a 1 to 2 week period.
30. Method according to any one of claims 21 to 29, wherein the bacterial species is administered in the amount of about 108 to about 10" live bacteria.
31.. Method according to any one of claims 21 to 30, wherein the antigen is administered at a dose of 4 to 10 μg.
32. Method according to claim 31 , wherein the antigen is administered at a dose of 8 μg-
33. Method according to any one of claims 21 to 32, wherein the antigen is administered intraperitoneally.
34. Method according to any one of claims 21 to 33, wherein the antigen is administered at the same time or shortly after administration of the bacterial species.
35. Method according to any one of claims 21 to 34, wherein the antigen is administered 1 day after the administration of 4 to 8 oral doses of bacteria.
36. Method according to any one of claims 21 to 35, wherein administration of the bacteria species is continued after administration of the antigen.
37. Method according to any one of claims 21 to 36, wherein about 8 oral doses of bacteria are administered.
38 Method according to any one of claims 21 to 37, wherein the bacteria are administered at 2 day intervals.
39. Method according to any one of claims 21 to 38, wherein IgE is measured in serum and wherein the serum is collected approximately 14 days after administration of the antigen.
40. Method according to any one of claims 21 to 39, wherein IgE antibody level is determined using an immunoassay.
41 . Bacterial species identified by the method according to any one of claims 21 to 40.
42. Composition including an effective amount of live bacterial species according to claim 41.
43. Composition according to claim 42 in form of a tablet or capsule.
44. Composition according to claim 42 in form of a food product
45. Pharmaceutical composition including an effective amount of live bacterial species according to claim 41 , together with a pharmaceutically acceptable carrier, adjuvant, solvent or excipient.
46. Bacterial species according to claim 41, which is L. acidophilus.
47. Composition according to any one of claims 42 to 45, wherein the bacterial species is L. acidophilus.
48. Method according to any one of claims 2 to 18, wherein the allergy is selected from the group consisting of asthma, eczema, hayfever and food allergy.
49. Method of assessing efficacy of treatment with live probiotic bacteria or with a composition having live probiotic bacteria, including the steps of: a measuring the level of salivary immunoglobulin subclass in a sample obtained before commencement of treatment, b measuring the level of salivary immunoglobulin subclass in a sample obtained after commencement of treatment c comparing the levels of salivary immunoglobulin subclass in a) and b), wherein the change in immunoglobulin subclass level is indicative of effective treatment.
50. Method according to claim 49, wherein the immunoglobulin subclass is IgGl or IgG2.
51. Method of assessing efficacy of treatment with live probiotic bacteria or with a composition having live probiotic bacteria, including the steps of: a measuring the level of IL-4 or IFN-γ in a sample obtained before commencement of treatment, b measuring the level of IL-4 or IFN-γ in a sample obtained after commencement of treatment c comparing the levels of IL-4 or IFN-γ in a) and b), wherein the change in IL-4 or IFN-γ level is indicative of effective treatment.
EP00975685A 1999-11-19 2000-11-20 Compositions and methods for treatment of allergic disorders Withdrawn EP1229930A4 (en)

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Families Citing this family (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ID29150A (en) 1999-01-15 2001-08-02 Entpr Ireland Cs USE OF LACTOBACILLUS SALIVARIUS
FI110668B (en) 2000-06-20 2003-03-14 Aboatech Ab Oy Use of probiotics for primary prophylaxis of atopic diseases
PE20030283A1 (en) * 2001-07-26 2003-05-01 Alimentary Health Ltd LACTOBACILLUS CASEI STRAINS
GB0124580D0 (en) 2001-10-12 2001-12-05 Univ Reading New composition
JP4212838B2 (en) 2002-06-26 2009-01-21 カルピス株式会社 Antiallergic agent
EP1759597B1 (en) * 2003-03-13 2009-01-21 Kirin Holdings Kabushiki Kaisha Probiotic composition
JP4712289B2 (en) * 2003-08-26 2011-06-29 株式会社エイ・エル・エイ Immune promoting composition
JP4591810B2 (en) * 2003-09-19 2010-12-01 日之出産業株式会社 Nasal composition for controlling hay fever allergic rhinitis
JP4850715B2 (en) * 2003-12-17 2012-01-11 エヌ．ブイ．・ヌートリシア Lactic acid producing bacteria and lung function
US7955834B2 (en) 2004-06-03 2011-06-07 Biogaia Ab Method for improved breast milk feeding to reduce the risk of allergy
TWI356680B (en) * 2007-01-05 2012-01-21 Promd Biotech Co Ltd Anti-allergy lactic acid bacteria
CN101328468B (en) * 2007-06-21 2012-05-23 东宇生物科技股份有限公司 Antiallergic lactic acid bacteria
CN102657260B (en) * 2007-06-21 2013-08-21 东宇生物科技股份有限公司 Anti-allergic lactic acid bacteria
EP2065048A1 (en) * 2007-11-30 2009-06-03 Institut Pasteur Use of a L. casei strain, for the preparation of a composition for inhibiting mast cell activation
PL2244734T3 (en) 2008-02-01 2017-01-31 Murdoch Childrens Research Institute A method of inducing tolerance to an allergen
CA2808610C (en) 2010-08-10 2018-09-25 R.P. Scherer Technologies, Llc Process of manufacturing a stable softgel capsule containing microencapsulated probiotic bacteria
KR101311989B1 (en) * 2010-11-17 2013-09-26 주식회사한국야쿠르트 Products containing Lactobacillus casei HY7211 having immuno-regulatory and immuno-stimulatory functions as effective component
SG11202005863TA (en) * 2018-01-12 2020-07-29 Gi Innovation Inc Composition comprising probiotics and polypeptide having binding affinity for ige and use thereof
CN111296842B (en) * 2020-02-14 2021-01-05 赵一鸣 A kind of probiotic composition with anti-allergy effect
CN112375722B (en) * 2021-01-18 2021-04-13 山东中科嘉亿生物工程有限公司 Lactobacillus casei LC-12 for improving allergy, and product and application thereof
WO2024081219A1 (en) 2022-10-14 2024-04-18 Csp Technologies, Inc. Container and method for storing and stabilizing moisture sensitive products
CN117085046A (en) * 2023-10-20 2023-11-21 潍坊君薇生物科技有限责任公司 Lactobacillus acidophilus LS001 metaplasia and application thereof
CN117887643B (en) * 2024-03-14 2024-06-14 微康益生菌(苏州)股份有限公司 An anti-allergic probiotic and its application

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5522641A (en) 1978-08-07 1980-02-18 Kyokuto Shibousan Kk Antiallergic agent
JPS55122723A (en) 1979-03-14 1980-09-20 Kyokuto Shibousan Kk Drug for asthma
JPH07265064A (en) 1993-11-23 1995-10-17 Taketoshi Yamada Composition for improving enterobacterial flora
JPH092959A (en) 1995-06-16 1997-01-07 Yakult Honsha Co Ltd IgE antibody production inhibitor and antiallergic agent

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0904784A1 (en) * 1997-09-22 1999-03-31 N.V. Nutricia Probiotic nutritional preparation
JPH11199495A (en) * 1998-01-10 1999-07-27 Nichinichi Seiyaku Kk Anti-allergic agent using enteric capsule
IT1298918B1 (en) * 1998-02-20 2000-02-07 Mendes Srl USE OF ARGININE DEIMINASE BACTERIA TO INDUCE APOPTOSIS AND / OR REDUCE AN INFLAMMATORY REACTION AND PHARMACEUTICAL COMPOSITIONS
JP4074006B2 (en) * 1998-06-24 2008-04-09 アサマ化成株式会社 Method for producing IgE antibody production inhibitor and food using the same

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5522641A (en) 1978-08-07 1980-02-18 Kyokuto Shibousan Kk Antiallergic agent
JPS55122723A (en) 1979-03-14 1980-09-20 Kyokuto Shibousan Kk Drug for asthma
JPH07265064A (en) 1993-11-23 1995-10-17 Taketoshi Yamada Composition for improving enterobacterial flora
JPH092959A (en) 1995-06-16 1997-01-07 Yakult Honsha Co Ltd IgE antibody production inhibitor and antiallergic agent

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
See also references of WO0137865A1
SHIDA K. ET AL.: "LACTOBACILLUS CASEI INHIBITS ANTIGEN-INDUCED IGE SECRETION THROUGH REGULATION OF CYTOKINE PRODUCTION IN MURINE SPLENOCYTE CULTURES", INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY, vol. 115, 1998, pages 278 - 287, XP008007352

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