HK1115067B - Immune function modulating agent - Google Patents

Description

Immune function regulator

Technical Field

The present invention relates to an agent for preventing and/or treating allergy (also referred to as allergy), and a food composition for preventing and/or treating allergy, which contains the agent.

Background

Allergic diseases such as bronchial asthma, allergic rhinitis, and atopic dermatitis have been increasing rapidly over the past several decades, and it is considered that at least about 1/5 of the population suffers from some allergic diseases. Many of conventional allergy remedies are symptomatic treatments, and from the viewpoint of an increase in the number of patients suffering from allergy and side effects associated with long-term use, more effective treatments are also required (non-patent document 1).

Lactic acid bacteria are microorganisms having extremely high safety because they produce a suitable flavor substance and also have a function of producing an antibacterial substance such as lactic acid or bacteriocin, and therefore they have been conventionally eaten in various places throughout the world through fermented milk and the like. In recent years, in a double-blind test placebo trial, administration of Lactobacillus rhamnosus GG strain (Lactobacillus rhamnosus ATCC53103 strain), which is one of lactic acid bacteria, has been shown to suppress the onset of atopic diseases at high risk to about half (non-patent document 2), and it can be said that the use of lactic acid bacteria is a simple and effective method for preventing and/or treating allergic reactions without side effects.

Heretofore, various agents for preventing and/or treating allergic reactions using lactic acid bacteria have been proposed. However, for example, in the case of lactic acid bacteria (Lactobacillus plantarum CCRC 12944 strain, Lactobacillus acidophilus CCRC 14079 strain) disclosed in patent document 1, the function of producing interferon- γ by cultured cells was evaluated when selecting lactic acid bacteria, but actually, the prevention and/or treatment effect of allergic reaction when ingested by animals was not studied. In addition, from the viewpoint of safety, probiotics (probiotics) isolated from the human body are preferable (non-patent document 3), but in the case of lactic acid bacteria (Lactococcus lactis subsp. lactis G50 strain, Lactobacillus paracasei KW3110 strain) described in patent document 2 and patent document 3, it is not considered that the strain is a Lactobacillus derived from human when selecting lactic acid bacteria. The lactic acid bacteria described in patent documents 2 and 4 (Lactococcus lactis G50 strain, Lactobacillus acidophilus CP1613 strain, Lactobacillus acidophilus L92 strain, Lactobacillus fermentum CP34 strain) have not been considered as probiotic bacteria when selecting lactic acid bacteria, i.e., resistant to bile acids in the environment at low pH in the stomach or in the intestinal tract (patent documents 2 and 4). In addition, the lactic acid bacteria disclosed in patent document 3 (Lactobacillus paracasei strain KW 3110) were selected using as an index the production of antigen-specific IgE antibodies of systemic immunity induced by administering an antigen into the abdominal cavity of a mouse (patent document 3). However, it is considered that the therapeutic effect on food allergy cannot be appropriately evaluated by using the production of antibodies by systemic immunity as an index. Food taken orally is recognized by the immune system of the intestinal tract itself, which is different from the systemic immune system, and is also called intestinal collateral lymphoid tissue (GALT), while being absorbed by the intestinal tract, and in some cases, oral immune tolerance is induced, and IgE antibody production is induced to cause food allergy. The pathogenesis of food-induced food allergy, which should induce immune tolerance, is not sufficiently understood, but at least both the intestinal immune system and the systemic immune system are involved in the pathogenesis of food allergy, and it is difficult to consider that the effect of preventing food allergy is reflected in the therapeutic effect of food allergy when the systemic immune system is used to evaluate the effect of preventing food allergy.

Thus, in the present situation, there are many places where improvement is required in the conventional lactic acid bacteria for preparing an intended allergy preventing and/or treating agent or an allergy preventing and/or treating food composition.

Patent document 1: japanese patent laid-open publication No. 2004-91491

Patent document 2: japanese patent laid-open publication No. 2004-18469

Patent document 3: japanese patent No. 3585487

Patent document 4: japanese patent laid-open publication No. 2004-26729

Non-patent document 1: akaxing-shine, Yuli-Zhengshi-Mei, Baichuantai (beautiful white-flower Tai Lang), アレルギ (topic of Chinese medicine における's nearest patient , latest medicine, 58(2), pp.7-14 (2003))

Non-patent document 2: kalliomaki M, Salminen S, Arvilomami H, Kero P, Koskinen P, Isolauri E, & Probiotics in primary preservation of atopic disease: a random simulated placebo-controlled tertiary, Lancet, 357(9262), pp.1076-1079(2001)

Non-patent document 3: naidu AS, Bidlack WR, Clemens RA, [ biological spectra of Lactic Acid Bacteria (LAB) ], Critical Reviews in Food Science and evaluation, 39(1), pp.13-126(1999)

Non-patent document 4: yizhen relates to (food アレルギ study) the sidetracking, brush-seeding immunization system における food アレルゲン's (blood sugar), Linshui research ジヤ - ナル, 24(5), pp.9-14(2001)

Disclosure of Invention

That is, the present invention has an object to select a lactic acid bacterium derived from the human intestine, which has an antiallergic activity typified by food allergy and has a high biological residue after administration, and to provide an agent for preventing and/or treating an allergic reaction using the lactic acid bacterium and/or a treated product of the lactic acid bacterium, and a food composition for preventing and/or treating an allergic reaction, which contains the agent.

The present invention has been made to solve the above problems, and the present inventors have conducted extensive selection operations by resetting the following criteria when selecting a lactic acid bacterium to be used in a target allergy preventing and/or treating agent. Specifically, the present inventors have conducted intensive studies on the selection of a lactic acid bacterium 273 strain belonging to the genus lactobacillus, which has been isolated from feces of adults alone, from the following strains: (1) high gastric acid/bile acid tolerance, (2) high IL-12 production promoting activity and Th1/Th2 balance improving effect on spleen cells derived from mice, (3) high inhibitory effect on antigen-specific IgE production induced by intraperitoneal administration of ovalbumin to BALB/C mice, (4) high inhibitory effect on antigen-specific IgE production induced by oral administration of food antigen (casein) to C57BL/6N mice, (5) high activation ability of natural killer cells, (6) high IL-12 production promoting activity and Th1/Th2 balance improving effect on spleen cells and mesenteric lymph node cells derived from mice immunized with ovalbumin, (7) high inhibitory ability of eosinophilia (eosinophila) induced by extraction of antigen from cedar pollen, and as a result, Lactobacillus plantarum OLL2809 strain (Lactobacillus gasseri 2809) as a probiotic lactic acid bacterium meeting these conditions, was found, The preservation number is as follows: NITE BP-72), and the present invention has been completed.

The effect described in (1) above is related to the biological residue after administration. The Lactobacillus gasseri OLL2809 strain (NITE BP-72) having such an effect is expected to be a substance having a high biological residue in the intestinal tract when administered as a viable bacterium to an agent for preventing and/or treating allergic reactions. The effects described in (2) to (7) above are related to enhancement of Th1 response or suppression of Th2 response in vivo. Allergic diseases are considered to be caused by disruption of the balance of Th1 and Th2 of T-cell synergies. The allergy-preventing and/or-treating agent or the allergy-preventing and/or-treating food composition of the present invention having the effects of (2) to (7) is useful for comprehensively improving the balance of Th1/Th2, which is inclined to Th2, to regulate immune function. Therefore, the agent for preventing and/or treating anaphylaxis of the present invention can exhibit the effect of preventing and/or treating anaphylaxis even when the strain Lactobacillus gasseri OLL2809 (deposited as NITE BP-72) used is dead, and can be expected to have a synergistic effect such as an effect of improving intestinal flora when administered as a viable bacterium. In particular, the effect described in (4) above is obtained by evaluation using a food allergy model, and the therapeutic effect of the agent for preventing and/or treating anaphylaxis according to the present invention on food allergy can be expected to be high.

Namely, the present invention provides the following.

An agent for preventing and/or treating allergic reaction, which comprises Lactobacillus gasseri as an active ingredient.

An agent for preventing and/or treating anaphylaxis containing lactobacillus processed product as effective component, wherein the lactobacillus processed product is at least one selected from culture, concentrate, paste, spray dried product, freeze dried product, vacuum dried product, roller dried product, liquid product, diluted product, and crushed product of lactobacillus gasseri.

The agent for the prophylaxis and/or treatment of allergic reaction according to any one of the above [1] to [2], wherein the prophylaxis and/or treatment of allergic reaction is carried out by inhibiting the production of antigen-specific IgE.

The agent for the prophylaxis and/or treatment of allergic reaction according to any one of the above [1] to [2], wherein the prophylaxis and/or treatment of allergic reaction is carried out by promoting IL-12 production and improving the balance Th1/Th 2.

The agent for the prophylaxis and/or treatment of allergic reaction according to any one of the above [1] to [2], wherein the prophylaxis and/or treatment of allergic reaction is carried out by activation of natural killer cells.

The agent for the prevention and/or treatment of allergic reaction according to any one of the above [1] to [2], wherein the prevention and/or treatment of allergic reaction is carried out by inhibiting the increase of eosinophils.

The agent for the prophylaxis and/or treatment of allergic reaction according to any one of the above [1] to [2], wherein the prophylaxis and/or treatment of allergic reaction is carried out by at least one selected from the group consisting of inhibition of production of antigen-specific IgE, promotion of IL-12 production, improvement of Th1/Th2 balance, activation of natural killer cells, and inhibition of increase of eosinophils.

The agent for the prophylaxis and/or treatment of allergic reaction according to any one of the above [1] to [7], wherein the Lactobacillus gasseri is Lactobacillus gasseri OLL2809 strain (Lactobacillus gasseri OLL2809) deposited under accession No. NITE BP-72.

A food composition for preventing and/or treating allergic reactions, which comprises an effective amount of the agent for preventing and/or treating allergic reactions according to any one of [1] to [8 ].

The food composition for preventing and/or treating allergic reactions according to the above [9], which is a food such as modified milk powder for infants, milk powder for infants and the like, a food such as milk powder for women in lactation and the like, a health functional food, a food for patients or fermented milk.

Use of the agent for preventing and/or treating allergic reaction according to any one of the above [1] to [8] for producing a food composition for preventing and/or treating allergic reaction.

The use according to the above [11], wherein the food composition for preventing and/or treating allergic reaction is a food such as modified milk powder for infants, a food such as milk powder for women in lactation, a health functional food, a food for patients, or fermented milk.

A method for preventing and/or treating allergy, characterized by administering Lactobacillus gasseri or a treated product thereof.

A method for producing a food composition for preventing and/or treating allergic reactions, characterized in that Lactobacillus gasseri or a culture, concentrate, paste, spray-dried product, freeze-dried product, vacuum-dried product, roller-dried product, liquid product, diluted product or crushed product thereof is used as a growth promoter, and a meat extract, fish meat extract, yeast extract or the like can be used.

Lactobacillus gasseri OLL2809 strain deposited under accession number NITE BP-72 (Lactobacillus gasseri OLL 2809).

A treated product of lactic acid bacterium, which is a treated product of the strain Lactobacillus gasseri OLL2809 deposited under the accession number NITE BP-72, wherein the treated product is at least one selected from the group consisting of a culture, a concentrate, a paste, a spray-dried product, a freeze-dried product, a vacuum-dried product, a drum-dried product, a liquid product, a diluted product and a crushed product of the lactic acid bacterium.

A food composition or a beverage containing the Lactobacillus gasseri OLL2809 strain according to [15] or the treated lactic acid bacterium according to [16 ].

A medicament comprising the Lactobacillus gasseri OLL2809 strain of [15] or the treated lactic acid bacterium of [16 ].

As confirmed in examples described below, the agent for preventing and/or treating an allergic reaction of the present invention has tolerance to a low pH environment and bile acids, has an activity of promoting IL-12 production, has an effect of improving Th1/Th2 balance, has an ability to block production of food antigen-specific IgE in serum, an ability to activate natural killer cells, and an ability to suppress eosinophilia, and therefore can provide a novel agent for preventing and/or treating an allergic reaction which is effective for preventing and/or treating various allergic reactions including food allergic reactions, and a food composition for preventing and/or treating an allergic reaction containing the agent.

Drawings

FIG. 1 shows IgE inhibition by oral administration of lactobacillus in gastric acid tolerance/bile acid tolerance to BALB/c-OVA in an allergy model. A control group (control), a test cell administration group, and a comparative control group (Lactobacillus crispatus JCM 1185) are shown^TL.plantarum JCM 1149^TLactobacillus gasseri JCM1131^T) The mean value of the concentration (AU) of the anti-OVA-specific IgE antibody ± standard deviation (n ═ 10). p < 0.05 (relative to control, Fisher's PLSD test).

FIG. 2 shows the effect of acid/bile acid tolerant Lactobacillus on the natural killer activity of mouse spleen cells. Mean values ± standard deviations (n ═ 4) of the cell-mediated disorder activities (%) of splenocytes obtained from the control, test cell administration, and positive control (lactobacillus rhamnosus l. rhamnosus ATCC53103) are shown.^*: p < 0.05 (multiple comparative tests with respect to Lactobacillus gasseri OLL2809, Dunnet).

FIG. 3 shows cytokine production from splenocytes and mesenteric lymph node cells prepared by intraperitoneal administration of BALB/c-OVA orally administered lactobacillus to mice model for allergic reaction. The mean ± standard deviation (n-10) of each cytokine in the control group (control), the comparative control group (lactobacillus crispatus JCM 1185T), and the test cell administration group (lactobacillus gasseri OLL2809, l gasseri MEP170413) are shown.^*: p < 0.05 (relative to control, Fisher's PLSD test).

Detailed Description

The present invention will be described in detail below.

Disclosed is an agent for preventing and/or treating allergic reactions, which contains a lactic acid bacterium as an active ingredient. The agent for preventing and/or treating allergic reactions of the present invention comprises Lactobacillus gasseri as a lactic acid bacterium. Any species can be used as long as it has an allergy preventing and/or treating effect, but a species having a high biological residue after in vivo administration is preferable. Specific examples of lactobacillus gasseri that can be used in the present invention include lactobacillus gasseri OLL2809 strain. This time, Lactobacillus gasseri OLL2809 strain (Lactobacillus gasseri OLL2809, accession number: NITE BP-72) which is a lactic acid bacterium having an allergy preventing and/or treating effect was selected from Lactobacillus 273 strain isolated from adult feces alone, and was characterized in that it showed a higher activity in any of the items by the following test: (1) a gastric acid/bile acid tolerance test, (2) an evaluation test of an IL-12 production promoting effect on mouse-derived spleen cells and an improvement effect on Th1/Th2 balance, (3) an evaluation test of an inhibitory effect on antigen-specific IgE production induced by intraperitoneal administration of ovalbumin to BALB/C mice, (4) an evaluation test of an inhibitory effect on antigen-specific IgE production induced by oral administration of a food antigen (casein) to C57BL/6N mice, (5) an evaluation test of activation ability of natural killer cells, (6) an evaluation test of an IL-12 production promoting effect and an evaluation test of a Th1/Th2 balance improving effect on spleen cells and mesenteric lymph node cells derived from mice immunized with ovalbumin, and (7) an evaluation test of an inhibition ability of increase in eosinophilia induced by cedar pollen. Therefore, by using the lactic acid bacterium, a novel agent for preventing and/or treating allergic reactions, which is effective for preventing and/or treating various allergic reactions including food allergic reactions, and a food composition for preventing and/or treating allergic reactions, which contains the agent, can be provided.

The present inventors deposited this strain in the patent microorganism collection center of the basic agency for product evaluation technology of independent administrative Law. The contents of the specific deposit are described below.

The present inventors deposited Lactobacillus gasseri OLL2809 strain at the patent deposit center of the basic institute of product evaluation technology, independent administrative Law. The contents of the specific deposit are described below.

(1) The name of the depository institution: independent administrative legal system person product evaluation technology basic organization patent microorganism collection center

(2) Contact address:292-Oak 0818 Ozugaku-Ozugazu-Shizu-Ozu-ken-Shizu かずさ Sickle Zuka

Telephone number 0438-20-5580

(3) The preservation number is as follows: NITE BP-72

(4) Representation for recognition: lactobacillus gasseri OLL2809

(5) The original preservation date: 2 months and 1 day in 17 years (2005)

(6) Date of transfer to deposit under the budapest treaty: 2006, 1 month and 18 days

Lactobacillus gasseri OLL2809 strain (accession number: NITE BP-72) is gram-positive bacillus, and the clone on Lactobacillus MRS agar Difco is round, light yellow and flat. The physiological characteristics include the same type of lactic acid fermentation (also called homolactic fermentation) and the development at 45 ℃, and the fermentation properties for glucose, mannose, fructose, galactose, sucrose, cellobiose, lactose, and trehalose.

As a medium for culturing Lactobacillus gasseri OLL2809 (accession number: NITE BP-72), a medium generally used in the culture of lactic acid bacteria can be used. That is, any medium may be used as long as it contains other nutrients such as nitrogen sources and inorganic substances in an appropriate amount in addition to the main carbon source. As the carbon source, lactose, glucose, sucrose, fructose, starch hydrolysate, molasses and the like can be used depending on the absorption ability of the bacteria used. As the nitrogen source, organic nitrogen-containing substances such as casein hydrolysate, whey protein hydrolysate, and soybean protein hydrolysate can be used. In addition, as the proliferation promoter, meat extract, fish meat extract, yeast extract, and the like can be used.

The culture is preferably carried out under anaerobic conditions, but may be carried out under microaerobic conditions by static culture using a liquid commonly used, or the like. The anaerobic culture may be performed by a known method such as a method of culturing under a carbon atmosphere, or may be performed by other methods. The culture temperature is preferably 30 to 40 ℃ in general, but other temperature conditions may be used as long as the temperature is a temperature at which the bacteria grow. The pH of the medium during culture is preferably maintained at 6.0 to 7.0, but other pH conditions may be used as long as the pH is a pH at which the bacteria grow. In addition, it can be cultured under batch culture conditions. The culture time is preferably 10 to 24 hours in general, but may be other culture times as long as the bacteria can grow.

Other lactobacillus gasseri that can be used in the present invention also exhibit the same characteristics as lactobacillus gasseri OLL2809 in terms of gram-positive bacilli and other physiological characteristics. Such Lactobacillus gasseri can be isolated from human feces and the like according to physiological characteristics by those skilled in the art. The effect of preventing and/or treating allergy can be confirmed by the examples of the test for evaluating the effect of promoting IL-12 production and the effect of improving Th1/Th2 balance, the test for evaluating the effect of suppressing antigen-specific IgE production (i.e., intraperitoneal administration and oral administration of antigen), the test for evaluating the ability of natural killer cells to activate, and the test for evaluating the ability of suppressing eosinophilia, specifically, the methods shown in the examples. The biological residue after in vivo administration can be confirmed by a gastric acid/bile acid tolerance test, specifically, by the method described in examples. The culture can also be performed by the same method as the above-described culture of Lactobacillus gasseri OLL 2809.

The allergy-preventing and/or-treating agent of the present invention may contain the aforementioned lactobacillus gasseri in various states, and may contain, for example, a suspension of lactic acid bacteria, a culture of lactic acid bacteria (cells, culture supernatant (containing a culture medium component)), a fermented lactic acid bacteria (lactic acid bacteria beverage, yogurt, etc.), and the like.

The allergy-preventing and/or-treating agent of the present invention may contain the aforementioned lactobacillus gasseri as it is, or may contain the lactobacillus gasseri as a treated product of lactobacillus gasseri subjected to some treatment. Examples of the treated lactic acid bacteria used in the agent for preventing and/or treating an allergic reaction of the present invention include lactic acid bacteria, lactic acid bacteria-containing products, concentrates of fermented milk, pastes, dried products (at least one selected from spray-dried products, freeze-dried products, vacuum-dried products, and roller-dried products), liquid products, diluted products, and crushed products. As the lactic acid bacteria, living bacteria, moist bacteria, dry bacteria, and the like can be suitably used. The microorganism may be a dead microorganism subjected to sterilization, that is, to heat sterilization, radiation sterilization, or disruption treatment. The extract can also be added into medicines and/or beverages and foods with biological specifications such as milk powder, and can be applied to various medicines and/or beverages and foods regardless of the forms of the medicines and/or beverages and foods.

The agent for preventing and/or treating allergic reactions of the present invention can be orally administered alone, or orally administered after mixing with other ingredients commonly used in pharmaceuticals or foods, or used in combination with other compounds or microorganisms having anti-allergic activity, and is effective for preventing allergic reactions and reducing (treating) allergic symptoms in humans and animals. Can be used for preventing and/or treating anaphylaxis. It has been determined that a shift in the Th1/Th2 balance toward the Th2 side is one of the causes of allergic diseases such as atopic dermatitis or allergic rhinitis (Hopkin, J.M.2002.the ise of atom anchors to infection. allergy 57: 5-9. Preschott, S.L., C.Macaubas, T.Smallacombe, B.J.Holt, P.D.Sly, and P.G.Holt.1999.development of environmental-specific T-cell memory in and normal chip. Lance 353: 196-200.Shirakawa, T.Enomoto, S.Shimizu, and J.M.Hopkin.1997. infection therapy and 275.79. science). The agent for preventing and/or treating anaphylaxis of the present invention has (1) an effect of increasing the production of IL-12, which is produced by dendritic cells and has a function of differentiating into Th 1-promoting cells, in cells obtained from an animal model for anaphylaxis; (2) since the effect of suppressing the production of the Th2 cytokine (IL-4) is expected, the effect of correcting the Th1/Th2 balance biased toward the Th2 side to the Th1 side can be expected. In addition, NK cells have the property of being activated by Th1 cytokine (IL-2) and IL-12, and the allergy preventing and/or treating agent of the present invention has been shown to have the NK cell activating effect, and from this point of view, the Th1/Th2 balance improving effect has also been confirmed. The allergy-preventing and/or-treating agent of the present invention has an inhibitory effect on eosinophilia, and it can be said that the agent has an effect on the prevention and/or treatment of allergy. Eosinophils are known as leukocytes which are increased as one of inflammatory cells in type I allergy, and are also increased in parasitic infections. As diseases in which eosinophils show high values, allergic rhinitis, allergic conjunctivitis, bronchial asthma, atopic dermatitis, food allergy, allergy to foreign proteins, parasitic infection, paroxysmal pulmonary eosinophilia syndrome, and the like are known (Howanitz JH, Howanitz PJ editors, river field monitoring, Laboratory Medicine bed resistance determination と interpretation , japanese patent publication, pp.583 (1995)). Furthermore, since IgE production is also increased in type I allergy, the present invention having an effect of inhibiting eosinophilia or IgE production is expected to have an effect of alleviating symptoms such as allergic rhinitis, allergic conjunctivitis, bronchial asthma, atopic dermatitis, food allergy, allergy to foreign proteins, and allergic reaction (allergic reaction). The type, symptoms and diseases of anaphylaxis to which the agent for preventing and/or treating anaphylaxis of the present invention can be applied are not particularly limited, and examples thereof include type I to type IV anaphylaxis, food anaphylaxis, pollinosis, atopic dermatitis, bronchial asthma, allergic conjunctivitis, allergic rhinitis, allergic gastroenteritis, anaphylactic reaction, drug anaphylaxis, urticaria, serum sickness, hemolytic anemia, contact dermatitis, myasthenia gravis, Goodpasture's syndrome (also referred to as Goodpasture's syndrome), glomerulonephritis (glomorulonephritis), and the like. The allergen is not particularly limited, and examples thereof include foods (wheat, barley, oat, rye, buckwheat, egg, milk, cheese, peanut, rice, corn, millet, barnyard grass, soybean, potato, yam, garlic, onion, carrot, parsley, celery, tomato, orange, peach, apple, kiwi, melon, strawberry, banana, walnut, sesame, mushroom, abalone, squid, salty salmon, shrimp, crab, salmon, mackerel, tuna, sardine, cod, squid, octopus, scallop, beef, chicken, pork, gelatin, etc.), animals (dog, cat, mouse, rat, pigeon, etc., or skin, body hair, feces, feather, etc.), insects (moth, butterfly, chironomid, hornet, bumblech, etc., and secretions of these insects, squash), mites, parasites (anis, fir, etc.), weeds (roundworm, etc.), ascarid, etc., koxiong, etc., grass (tree, pine, etc., and the like, Cypress, ragweed, gramineae, mugwort, sumac, alder, etc. or pollen, sap, etc. of these plants), mold, dust, house dust, rubber, metal, chemical, medicine, etc.

The agent for preventing and/or treating allergic reactions of the present invention may be used as it is, or may be used by concentrating a culture of lactic acid bacteria after completion of the culture to give a concentrate, or may be used by further drying the concentrate. The cell concentration is not particularly limited, but is preferably 4X 10 in the concentrated solution¹⁰The amount of the active ingredient is preferably 5X 10 in dry matter¹¹More than one per gram.

The amount of the agent for preventing and/or treating allergy of the present invention to be incorporated in a pharmaceutical product, a food or drink is not particularly limited, and may be, for example, 0.001 to 100% (w/w), preferably 0.01 to 100% (w/w), more preferably 0.1 to 100% (w/w), depending on the form, the symptom, the body weight, the use, and the like.

The daily dose of the drug, drink or food for the agent for preventing and/or treating anaphylaxis of the present invention is not particularly limited, and may be, for example, 0.1 to 10000mg/kg body weight, preferably 0.1 to 1000mg/kg body weight, because it varies depending on age, symptoms, body weight, use, etc.

The agent for preventing and/or treating allergic reaction of the present invention can be used in the form of any of drugs, beverages, and foods. For example, it is expected that various allergic reactions can be prevented and/or treated by direct administration as a pharmaceutical product, or by direct ingestion as a food for special use such as a food for specified health care or a food with nutritional function. In addition, it can be added to various foods and beverages (such as milk, processed milk, milk beverage, soft drink, fermented milk, yogurt, cheese, bread, biscuit, cracker, pizza, ice cream, candy, modified milk powder, liquid food, food for patients, food for infants, food such as milk powder for women in lactation period, and nutritional food) for ingestion.

The food or drink containing the agent for preventing and/or treating allergic reaction of the present invention may be used by mixing water, protein, sugar, lipid, vitamins, minerals, organic acids, organic bases, fruit juices, flavors, and the like. Examples of the protein include animal and plant proteins such as whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein concentrate, whey protein isolate, α -casein, β -casein, κ -casein, β -lactoglobulin, α -lactalbumin, lactoferrin, soybean protein, egg protein, meat protein, and hydrolysates thereof; and various milk-derived components such as butter, whey minerals, cream, whey, nonprotein nitrogen, sialic acid, phospholipids, and lactose. Examples of the saccharide include processed starch (soluble starch, British starch, oxidized starch, starch ester, starch ether, etc., in addition to dextrin), dietary fiber, and the like. Examples of the lipid include animal fats and oils such as lard, fish oil, and the like, and oils isolated therefrom, hydrogenated oils, and transesterified oils thereof; vegetable oils and fats such as palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, their separated oils, hydrogenated oils, and transesterified oils. Examples of the vitamins include vitamin a, carotenes, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, nicotinic acid, pantothenic acid, biotin, inositol, choline, and folic acid. Examples of the minerals include calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, and selenium. Examples of the organic acid include malic acid, citric acid, lactic acid, and tartaric acid. In the production of a food or drink containing the agent for preventing and/or treating allergic reaction of the present invention, these components may be either synthetic products or natural products, and/or a food containing a large amount of these components may be used as a raw material. These components may be used in combination of 2 or more.

When the allergy-preventing and/or-treating agent of the present invention is used in the production of a food composition or a pharmaceutical, the production method can be carried out by a method known to those skilled in the art. Those skilled in the art can appropriately combine the step of mixing the Lactobacillus gasseri cells or treated product of the present invention with other components, the molding step, the sterilization step, the fermentation step, the baking step, the drying step, the cooling step, the granulation step, the packaging step, and the like, and can produce desired foods or medicines.

When the Lactobacillus gasseri of the present invention is used for the production of various dairy products, desired dairy products can be produced by a method known to those skilled in the art. When an example of yogurt is given, yogurt can be produced through a step of preparing a starting material using Lactobacillus gasseri of the present invention, a step of adding the starting material to pretreated milk and culturing the milk, a cooling step, a seasoning step, a filling step, and the like. In the case of cheese, for example, the cheese can be produced by a step of adding the lactobacillus gasseri of the present invention as a starting material to milk subjected to a pretreatment such as sterilization and the like to lactic-ferment the milk, a step of adding rennet to produce cheese curd, a curd cutting step, a whey discharging step, a salting step, a ripening step, and the like. Alternatively, in the production of the various dairy products, other lactic acid bacteria may be used as a starting material in place of the Lactobacillus gasseri of the present invention, and then the cells or treated products of the Lactobacillus gasseri of the present invention may be added in the production process.

When the agent for preventing and/or treating anaphylaxis of the present invention is used as a pharmaceutical, it can be administered in various forms. Examples of the form of the drug include oral administration by tablets, capsules, granules, powders, syrups, and the like. These various preparations can be prepared by using known adjuvants which can be generally used in the field of pharmaceutical preparation technology, such as excipients, binders, disintegrants, lubricants, deodorants, dissolution aids, suspensions, and coating agents, in a main agent according to a conventional method.

In addition, all prior art documents cited in the present specification are incorporated herein by reference.

Examples

The present invention will be described below with reference to examples, but the present invention is not limited to these examples.

[ example 1] (gastric acid resistance test and bile acid resistance test)

Lactobacillus 273 strain isolated from human feces was subjected to the test.

1) Gastric acid resistance test

The lactic acid bacteria cells to be tested were washed 2 times with physiological saline, and 1ml of the lactic acid bacteria suspension was added to 9ml of filter-sterilized artificial gastric juice [ NaCl (0.2%), pepsin (1: 5000, Tokyo chemical industry) (0.35%) ] (pH 2). After aerobic culture at 37 ℃ for 2 hours, 1ml of the artificial gastric juice containing the lactic acid bacteria was taken out and added to 9ml of a phosphate buffer (67mM, pH 6.5) to stop the reaction. The number of viable bacteria before and after contact with the artificial gastric juice was measured using lactobacillus MRS agar (DIFCO) to determine the survival rate (%) as the gastric acid resistance (%).

2) Bile acid tolerance test

10. mu.l of lactic acid bacteria pre-cultured (activated ) (37 ℃ C., 18 hours) with Lactobacillus MRS broth 2 times were inoculated into 5ml of Lactobacillus MRS broth containing 0.9% bovine biliary powder (DIFCO) and anaerobically cultured at 37 ℃. After 18 hours of culture, the turbidity (OD650) of the medium at a wavelength of 650nm was measured, and this value was used as an index of bile acid tolerance.

The results of the gastric acid resistance test and the bile acid resistance test are shown in table 1. As a result of examining gastric acid tolerance and bile acid tolerance of 273 strains of lactobacillus, the following tendency was found: strains with high gastric acid tolerance, which have low bile acid tolerance; the bacterial strain with high bile acid tolerance has low gastric acid tolerance. And through strain separation and observation, the gastric acid tolerance is high in Lactobacillus gasseri, and the bile acid tolerance is high in Lactobacillus plantarum. Bile acid tolerance is 1.0 or less in most strains. As strains having high bile acid tolerance, Lactobacillus plantarum 6 strain, Lactobacillus crispatus 2 strain, Lactobacillus amylovorus (Lactobacillus amylovorus)2 strain, Lactobacillus casei (Lactobacillus casei)1 strain, and Lactobacillus brevis (Lactobacillus brevis)1 strain were selected. In addition, gastric acid tolerance is 0.5% or less in most strains. Among strains having high gastric acid tolerance (0.5% or more), lactobacillus gasseri 8 strain was selected as a strain having relatively high bile acid tolerance (0.1 or more).

TABLE 1

Table 1 shows the results of gastric acid tolerance (%), bile acid tolerance (OD650) of the selected lactobacillus 20 strain.

Example 2 preparation of lyophilized lactic acid bacteria

For 20 strains of lactobacillus selected in example 1, pre-cultured 2 times (37 ℃, 18 hours) with lactobacillus MRS broth. 1% of the cells obtained by the preculture were inoculated into the same medium and cultured at 37 ℃ for 18 hours. After collecting the bacteria, the cells were washed 2 times with physiological saline and 1 time with sterilized distilled water. Heated at 75 ℃ for 60 minutes for sterilization and freeze-dried. The freeze-dried powder was used in the following in vitro test and animal administration test.

[ example 3] (test for promoting IL-12 production from mouse-derived spleen cells and evaluating Th1/Th2 balance improving Effect)

The gastric acid-resistant, bile acid-resistant lactobacillus 20 strain selected in example 1 was evaluated by the following method.

To 6-week-old male BALB/c mice (n-4, Japanese SLC), 20. mu.g of ovalbumin (hereinafter, also referred to as OVA: Wako pure chemical industries) and 2mg of aluminum hydroxide (Wako pure chemical industries) were administered intraperitoneally, and the spleens were removed 8 days later. The red blood cells-removed splenocytes were suspended in 10% FCS-RPMI1640 medium (Gibco Co.) containing 1. mu.g/ml of the lactic acid bacteria heat-treated cells prepared in example 2 and 100. mu.g/ml of OVA to 2.5X 10⁶Per ml and at 5% CO₂Culturing in an incubator for 6 days. As a comparative control, a standard strain of Lactobacillus (Lactobacillus plantarum JCM 1149) was used^TLactobacillus gasseri JCM1131^TLactobacillus crispatus JCM1185^TLactobacillus amylovorus JCM 1126^T) And standard strains of other bacteria (Bifidobacterium bifidum JCM 1255)^T(Bifidobacterium bifidum JCM 1255^T) Bifidobacterium longum JCM 1217^T(Bifidobacterium longum JCM 1217^T) Lactobacillus delbrueckii subsp lactis JCM 1248^T(Lactococcus delbrueckii subsp.lactis JCM 1248^T) Enterococcus faecalis IFO 3971(Enterococcus faecalis IFO 3971), Bacteroides vulgatus JCM 5826^T(Bacteroides vulgatus JCM 5826^T) Escherichia coli JCM 1649^T(Escherichiacoli JCM 1649^T). Among the above strains, the strain described as JCM in the strain name was a standard strain obtained from the development room for microbial materials at the center of the institute of independent administrative sciences and physicochemical research, the strain described as IFO in the strain name was a standard strain obtained from the research institute of fermentation by the treasury corporation, and the strain wasThe strain named MEP is a strain owned by Mingzhi Dairy Co. In addition, a product prepared in the same manner as described above except that no microorganism was added was used as a control. Culture supernatants were assayed for IFN-. gamma., IL-12 and IL-4 by ELISA (BD OptEIATM ELISA set, Becton Dickinson). The significance difference relative to the control group was tested at a significance level (significant level) of 5% by Student's t-test. The results are shown in Table 2.

TABLE 2

^*: p < 0.05 (relative to control, Student's t-test)

Table 2 shows the effect of the Lactobacillus-strain 20 selected in example 1 on the production of cytokines (IL-12, IFN-. gamma., IL-4) by splenocytes. The mean value. + -. standard deviation of each measurement value is shown.^*: p < 0.05 (relative to control, Student's t-test)

With Lactobacillus plantarum, Lactobacillus gasseri, etc. as the center, 13 out of 30 strains strongly promoted IL-12 production significantly (p < 0.05). However, the IL-12-inducing activity was very potent and completely inactive in the same species, and was strain-specific regardless of the species. In addition, positive correlation (n-31, r-0.8047, p < 0.01) and negative correlation (n-31, r-0.6544, p < 0.01) were observed between IFN- γ and IL-4 and the amount of IL-12 produced, respectively. That is, the strain strongly inducing IL-12 production also had a high IFN-. gamma.production promoting effect and a high IL-4 production inhibiting effect, and the balance Th1/Th2 was improved. From the above results, it was found that specific strains belonging to Lactobacillus gasseri, Lactobacillus plantarum and the like including Lactobacillus gasseri MEP170409 (Lactobacillus gasseri OLL2809 (accession number: NITE BP-72)) had strong IL-12 production promoting activity and Th1/Th2 balance improving effect.

Thus, 4 strains of Lactobacillus plantarum MEP170402, Lactobacillus gasseri MEP170407, Lactobacillus gasseri MEP170409 (Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72)), and Lactobacillus gasseri MEP170413, which had high IL-12 production promoting activity and improved Th1/Th2 balance, were selected.

Example 4 (test for evaluating the inhibitory Effect of antigen-specific IgE production induced by intraperitoneal administration of ovalbumin to BALB/c mice)

The lactobacillus 4 strain selected in example 3 was evaluated by the following method.

Male BALB/c mice (groups n ═ 10, japan SLC) at 6 weeks of age were intraperitoneally administered with 0.2 μ g/g-body weight OVA (and Wako pure chemical industries) and 0.1 mg/g-body weight aluminum hydroxide (and Wako pure chemical industries) (primary immunization). After 2 weeks OVA and aluminium hydroxide were immunised twice under the same conditions. The mice were allowed to freely take MF feed containing 0.1% of each of the freeze-dried powders of lactic acid bacteria prepared in example 2 (Oriental yeast industry) within 21 days after 1 week from the primary immunization to the secondary immunization. The lactic acid bacterium was Lactobacillus crispatus JCM1185, which had low IL-12-inducing activity in the results of example 3^TLactobacillus plantarum JCM 1149^TLactobacillus gasseri JCM1131 with high IL-12 inducing activity^TAs comparative controls for lactic acid bacteria, the inhibitory effects against the production of OVA-specific IgE by Lactobacillus plantarum MEP170402, Lactobacillus gasseri MEP170407, Lactobacillus gasseri MEP170409 (Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72)), and Lactobacillus gasseri MEP170413 were examined. The control was MF feed only. Blood was collected 1 week after the second immunization, and the concentration of anti-OVA-specific IgE antibody in serum was measured by ELISA. As for the concentration of anti-OVA-specific IgE antibody, the concentration of anti-OVA-specific IgE contained in serum obtained by intraperitoneally administering 20. mu.g of OVA and 2mg of aluminum hydroxide at 2-week intervals to 6-week-old male BALB/c mice (Japanese SLC) and further 1 week later was defined as 10000 AU. The concentration of anti-OVA-specific IgE was determined by varying the method of Ito et al (Ito, K., K.Inagaki-Ohara, S.Murosaki, H.Nishimura, T.Shimokata, S.Torrii, T.Matsuda, and Y.Yoshikai, Murine modelof IgE production with immunogenic Th2-response by feeding protein antigen with out added mutations, Eur.J.Immunol.27: 3427-3437(1997), using biotinylated rat anti-mouse IgE monoclonal antibodies (BD biosciences) and streptavidin-horseradish peroxidase (BDbiosciences). Results were subjected to One-way Analysis of variance (One-way Analysis of variance), and further to multiple tests at a significance level of 5% by Fisher's PLSD test. The results are shown in FIG. 1.

Lactobacillus crispatus JCM1185 compared to a control^TAnd Lactobacillus plantarum JCM 1149^TNo difference was observed. Lactobacillus plantarum MEP170402, which showed a higher IL-12 induction effect in vitro, was also not different from the control. Among the strains tested, Lactobacillus gasseri OLL2809 (accession number: NITE BP-72) showed the strongest inhibitory effect on the production of anti-OVA-specific IgE, and a significant decrease (p < 0.05) was observed compared with the control. In addition, Lactobacillus gasseri JCM1131^TAnd lactobacillus gasseri MEP170407 showed a tendency to inhibit antigen-specific IgE induction (p 0.072, p 0.064, respectively). In vivo, lactobacillus plantarum does not show an inhibitory effect on IgE production, whereas lactobacillus gasseri is highly effective. As is clear from the above, the L2809 strain of Lactobacillus gasseri (accession number: NITE BP-72) has a strong inhibitory effect against the production of OVA-specific IgE.

Example 5 (test for evaluating the inhibitory Effect of antigen-specific IgE production induced by oral administration of food antigen to C57BL/6N mice)

The gastric acid-resistant and bile acid-resistant lactobacillus 20 strains selected in example 1 were evaluated by the following methods.

A 3-week-old female C57BL/6N mouse (control N20, test cell administration group N10, CLEA, japan) was allowed to freely take a feed (test cell administration group) having a composition shown in table 3, to which 0.1% of each of the freeze-dried powders of lactic acid bacteria prepared in example 2 was added, for 2 weeks. The control was given to a feed of the composition shown in table 3. All groups were given MF feed (Oriental yeast) 7 weeks later. Further, the test cell administration group was allowed to freely take 2 weeks of the feed having the composition shown in table 3 to which 0.1% of each of the freeze-dried powders of lactic acid bacteria prepared in example 2 was added, and the control group was allowed to freely take 2 weeks of the feed having the composition shown in table 3. After completion of the raising, total blood collection was performed, and an acute allergic reaction in the skin of a rat (hereinafter, also referred to as PCA (passive skin allergy) reaction) test was performed using the serum of each mouse, and the positive rate of anti-casein specific IgE was compared between the control and test cell administration groups.

TABLE 3

The AIN-76 vitamin mixture and AIN-76 mineral mixture described in table 3 refer to vitamin mixtures and mineral mixtures blended in AIN-76, which is a standard refined feed for nutritional studies of mice or rats published in 1977 by the american national institute for nutrition (AIN).

[ determination of IgE production-inhibitory Effect by PCA reaction ]

The PCA reaction was performed by injecting the serum of mice into the skin of a 7-week-old male SD rat (Japanese SLC) 25. mu.l each after unhairing the back. After 24 hours of intradermal injection of mouse serum, 0.5ml of a solution containing 2mg/ml of casein and 1% Evans Blue (and Wako pure chemical industries) was injected into the tail vein of the rat. Further, after 30 minutes of intravenous injection of casein and evans blue, the presence or absence of leakage of evans blue pigment under the back skin was visually judged, and a person with leakage of visible pigment was judged to be positive for PCA reaction. The PCA reaction positive numbers in the respective groups were determined, and the significance test of the PCA reaction positive numbers of the control group and the test cell administration group was performed at a significance level of 5% by Chi-square test. The results are shown in Table 4.

TABLE 4

^*: p < 0.05 (relative to control, chi fang test)

Table 4 shows the effect of oral administration of the Lactobacillus 20 strain selected in example 1 on IgE production in food-allergic mice. Shows the number of PCA reaction positives/number of tests.^*: p < 0.05 (relative to control, chi-square test).

In the control group, 18 out of 20 were positive. On the other hand, 5 of the group administered with Lactobacillus gasseri OLL2809 (accession number: NITE BP-72) and 10 positive numbers of Lactobacillus amylovorus MEP 170417 were significantly inhibited (p < 0.05). No significant effect was seen in the other strains.

As is clear from the above, the Lactobacillus gasseri OLL2809 (accession number: NITE BP-72) strain had a higher inhibitory effect on the production of antigen-specific IgE when orally administered to food allergy model mice by oral administration of casein, as compared to other strains.

[ example 6] (evaluation test for activation ability of Natural killer cell)

The gastric acid-resistant and bile acid-resistant lactobacillus 20 strains selected in example 1 were evaluated by the following methods.

Spleen cells were collected from 10-week-old female BALB/c mice (Japanese SLC). As the target cells, YAC-1 cell lines (cell lines showing high sensitivity to natural killer cells) were used. As fluorescent markers for YAC-1 cells, 1X 10⁶Calcein (Dojindo) was added to each ml of the cells at a final concentration of 20. mu.M, and the cells were left to stand for 1 hour and washed. 5X 10 cells were added to a 96-well plate in an amount of 100. mu.l per well of spleen cell fluid⁵To each of the lactic acid bacteria, 0.20. mu.g of the lyophilized powder of each of the lactic acid bacteria prepared in example 2 (final concentration: 1.3. mu.g/ml) was added in an amount of 50. mu.l, and the mixture was cultured overnight. Then, 1X 10 of the above-mentioned fluorescently labeled YAC-1 cells were added in a volume of 50. mu.l⁴One (splenocytes: YAC-1 cells at a ratio of 50: 1) and cultured for 3 hours. The control wells were prepared in the same manner except that no freeze-dried powder of lactic acid bacteria was added to the splenocytes. In addition, as a positive control, lactobacillus rhamnosus ATCC53103 strain (lactobacillus rhamnosus GG strain) was used. After the incubation, the fluorescence intensity (excitation: 485nm, emission: 535nm) in the supernatant was measured, and the value was used as an experimental measurement value. Furthermore, the fluorescence intensities in the culture supernatants obtained by culturing splenocytes alone or YAC-1 cells alone were determined as the spleen cell spontaneous measurement value and YAC-1 spontaneous measurement value, respectively. In addition, the fluorescence intensity at the time of destroying cells after adding Triton X100 (Wako pure chemical industries) after culturing YAC-1 cells alone was used as a total measurement value. These values were substituted into the following formula to calculate the cell-damaging activity. Statistical treatment the Dunnet multiple comparison test was performed, and the significance test was performed on Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72) strain at a significance level of 5%. The results are shown in FIG. 2.

Math formula (1)

In the above formula, Test represents a Test measurement value, Total represents a Total measurement value, S represents a spleen cell spontaneous measurement value, and Y represents a YAC-1 spontaneous measurement value.

Among the 20 strains studied, the cell-barrier activity was particularly high when strains belonging to Lactobacillus plantarum or Lactobacillus gasseri were added. In addition, the sample containing Lactobacillus gasseri OLL2809 (accession number: NITE BP-72) strain showed significantly (p < 0.05) high cell-damaging activity compared to the positive control sample.

As is clear from the above, the Lactobacillus gasseri OLL2809 (accession number: NITE BP-72) strain showed a high natural killer cell activation ability in a system using spleen cells.

From the above, it was found that Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72) strain as a lactic acid bacterium showed higher activity in the following tests: the test was conducted on (1) gastric acid/bile acid tolerance, (2) an evaluation test of the promotion effect of IL-12 production from mouse-derived spleen cells and the improvement effect of Th1/Th2 balance, (3) an evaluation test of inhibition of antigen-specific IgE production induced by intraperitoneal administration of ovalbumin to BALB/C mice, (4) an evaluation test of ability of inhibition of antigen-specific IgE production induced by oral administration of food antigen to C57BL/6N mice, and (5) an evaluation test of the activation ability of natural killer cells.

Example 7 (evaluation test of Th1/Th2 balance-improving Effect of splenocytes and mesenteric lymph node cells in BALB/c mice administered OVA intraperitoneally)

The lactobacillus 2 strain selected in example 3 was evaluated by the following method.

Male BALB/c mice (groups n ═ 10, japan SLC) at 6 weeks of age were intraperitoneally administered with 0.2 μ g/g-body weight OVA (and Wako pure chemical industries) and 0.1 mg/g-body weight aluminum hydroxide (and Wako pure chemical industries) (primary immunization). After 2 weeks OVA and aluminium hydroxide were immunised twice under the same conditions. The mice were allowed to freely take MF feed containing 0.1% of each of the freeze-dried powders of lactic acid bacteria prepared in example 2 (Oriental yeast industry) within 21 days after 1 week from the primary immunization to the secondary immunization. Splenocytes and mesenteric lymph node cells were prepared from each group of mice 1 week after the secondary immunization. The spleen cells were removed from the red blood cells at 2.5X 10⁶Per well concentration was seeded in 24-well plates with mesenteric lymph node cells at 1.25X 10⁶The concentration per well was seeded in 48-well plates. Each cell was cultured in 10% FCS-RPMI1640 medium (Gibco, 1 ml/well) containing 100. mu.g/ml OVA at a concentration of 5% CO₂The culture was carried out in an incubator at 37 ℃ for 2 days or 6 days. IL-12(p70) in the culture supernatant after 2 days of culture and IFN-. gamma.and IL-4 concentrations in the culture supernatant after 6 days of culture were measured by ELISA (BD OptEIATM ELISA kit, Becton Dickinson), respectively. Results when single-factor analysis of variance was performed and the significance was confirmed to be significant at a significance level of 5%, the results were further tested by Fisher's PLSD test at a significance level of 5%Multiple tests were performed.

Lactic acid bacteria Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72), Lactobacillus gasseri MEP170413 and Lactobacillus crispatus JCM1185 with low IL-12-inducing activity as a comparative control, which were selected by the results of example 3, were used^T. The control was MF feed only. The results are shown in FIG. 3.

For IL-12 production from spleen cells (p70), a significant increase (p < 0.05) was observed in the group to which Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72) strain was administered, as compared with the control. In addition, a significant (p < 0.05) decrease in the amount of IL-4 produced from spleen cells and mesenteric lymph node cells in the group to which Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72) strain was administered was also observed, as compared with the control.

In Lactobacillus gasseri MEP170413, a significant (p < 0.05) increase in IL-12 production from splenocytes was seen (p70), but IL-4 production was not affected in both splenocytes and mesenteric lymph node cells. Lactobacillus crispatus JCM1185^TNo changes were seen in cytokine production for these cells. As is clear from the above, L.gasseri OLL2809 (accession number: NITE BP-72) strain has an effect of improving the balance of Th1/Th2 between spleen cells and mesenteric lymph node cells when orally administered to BALB/c mice administered with OVA intraperitoneally.

(test for evaluating the inhibitory Effect of eosinophilia Using Cunninghamia lanceolata pollen extract antigen-induced eosinophilia model)

Japanese SLC mice (n 9 or 10/group) 7 weeks old female BALB/c mice were subcutaneously administered with 0.2ml of a Japanese cedar pollen-extracted antigen solution (containing 0.4. mu.g/ml Cry j 1) per one on the dorsal side on the 0 th, 1 th, 6 th, 8 th, and 14 th days from the start of the experiment, and were sensitized with an antigen. On day 20, the antigen solution extracted from cedar pollen was administered in the same manner to induce an increase in eosinophils. The negative control group was not sensitized, and only induced. For each group, water (control) or the Lactobacillus gasseri OLL2809 (accession No.: NITE BP-72) strain prepared in example 2 was forcibly administered to the stomach at various doses (0.5, 1.0, 2.0mg) every day for 21 days from the start of the experiment. The composition of the groups is as follows. On day 21, 5mL of phosphate buffered saline containing 1% FCS was intraperitoneally injected, and abdominal cells were collected. The total cell concentration of the abdominal cells was measured by an automatic hemocytometer (F-800, Sysmex Co.), and then an abdominal cell smear was prepared by a collection cell centrifuge (Cytospin3, Shandon Co.). Peritoneal cells were stained with a Hemacolor rapid staining kit (Merck) and the number of eosinophils was measured by a microscope. The eosinophil inhibitory rate was determined by the following equation.

Math formula (2)

TABLE 5

[0130]In the control group, the antigen was sensitized and induced by extraction of cedar pollen, and the total cell concentration, the eosinophil concentration and the ratio of eosinophils (number of eosinophils/total number of cells) in the recovered abdominal cavity were significantly increased (p < 0.01) as compared with the negative control group. In the group to which the L2809 strain of Lactobacillus gasseri (accession number: NITE BP-72) was administered, the proportion of eosinophils was significantly suppressed (p < 0.05 or 0.01) compared to the control group at any dose. In addition, no significant difference in total cell concentration of the peritoneal fluid was observed in all the groups subjected to sensitization and induction, and only the 2.0 mg-administered group showed a significantly low value of eosinophil concentration (p < 0.05) and significantly inhibited the increase of eosinophils (p < 0.05) by about 44% as compared with the control group.

From the above results, it was found that the OLL2809 strain inhibited the increase of eosinophils by oral administration to mice of a hypereosinophilia model.

TABLE 6

Mean ± standard deviation, n-9 for control group only, and n-10 for others.^*、^**: p < 0.05, 0.01 (Dunnett multiple comparison test relative to control). Cell concentration represents the concentration in the peritoneal cavity recovery fluid.

The present invention can provide a novel agent for preventing and/or treating allergic reactions. A food composition for preventing and/or treating allergic reactions is provided by adding an effective amount of the agent to infant formula milk powder, health functional food or patient food.

Claims (17)

1. An agent for preventing and/or treating allergic reactions, which comprises as an active ingredient a Lactobacillus gasseri OLL2809 strain deposited under accession No. NITE BP-72.

2. An agent for preventing and/or treating allergic reaction, which comprises a treated product of Lactobacillus gasseri OLL2809 strain deposited under the accession number NITE BP-72 as an active ingredient, wherein the treated product of the lactic acid bacterium is at least one selected from the group consisting of a culture, a concentrate, a paste, a spray-dried product, a freeze-dried product, a vacuum-dried product, a roller-dried product, a liquid product, a diluted product and a crushed product of the lactic acid bacterium.

3. The agent for the prophylaxis and/or treatment of allergic reaction according to any one of claims 1 to 2, wherein the prophylaxis and/or treatment of allergic reaction is carried out by inhibiting the production of antigen-specific IgE.

4. The agent for the prophylaxis and/or treatment of allergic reaction according to any one of claims 1 to 2, wherein the prophylaxis and/or treatment of allergic reaction is carried out by promoting IL-12 production and improving the balance Th1/Th 2.

5. The agent for the prophylaxis and/or treatment of allergic reaction according to any one of claims 1 to 2, wherein the prophylaxis and/or treatment of allergic reaction is carried out by activation of natural killer cells.

6. The agent for the prevention and/or treatment of allergic reaction according to any one of claims 1 to 2, wherein the prevention and/or treatment of allergic reaction is carried out by inhibiting the increase of eosinophils.

7.The agent for the prevention and/or treatment of allergic reactions according to any one of claims 1 to 2, wherein the prevention and/or treatment of allergic reactions is performed by at least one of inhibiting the production of antigen-specific IgE, promoting the production of IL-12 and improving the Th1/Th2 balance, activating natural killer cells, and inhibiting the increase of eosinophils.

8. A food composition for preventing and/or treating allergic reactions, which comprises an effective amount of the agent for preventing and/or treating allergic reactions according to any one of claims 1 to 7.

9. The food composition for preventing and/or treating allergic reactions according to claim 8, which is formulated milk powder for infants, milk powder for lactating women, health functional food, food for patients, or fermented milk.

10. Use of the agent for preventing and/or treating allergic reaction according to any one of claims 1 to 7 for the production of a food composition for preventing and/or treating allergic reaction.

11. The use according to claim 10, wherein the food composition for preventing and/or treating allergic reactions is modified milk powder for infants, milk powder for lactating women, health functional food, food for patients, or fermented milk.

12. Use of the strain Lactobacillus gasseri OLL2809 deposited under accession number NITE BP-72 for the preparation of an allergy preventive and/or therapeutic agent.

13. A process for producing a food composition for preventing and/or treating allergic reactions, which comprises using Lactobacillus gasseri OLL2809 strain deposited under accession number NITE BP-72 or at least one treated product selected from the group consisting of a culture, a concentrate, a paste, a spray-dried product, a freeze-dried product, a vacuum-dried product, a roller-dried product, a liquid product, a diluted product and a crushed product thereof.

14. Lactobacillus gasseri OLL2809 strain deposited under accession number NITE BP-72 (Lactobacillus gasseri OLL 2809).

15. The treated product of lactic acid bacteria is a treated product of the strain Lactobacillus gasseri OLL2809 deposited under the accession number NITE BP-72, and is at least one selected from the group consisting of a culture, a concentrate, a paste, a spray-dried product, a freeze-dried product, a vacuum-dried product, a roller-dried product, a liquid product, a diluted product, and a crushed product of the lactic acid bacteria.

16. A food composition or a beverage containing the Lactobacillus gasseri OLL2809 strain according to claim 14 or the treated lactic acid bacterium according to claim 15.

17. A pharmaceutical composition comprising the Lactobacillus gasseri OLL2809 strain of claim 14 or the treated lactic acid bacterium of claim 15.