JPWO2008023664A1 - Bacterial strain having antiallergic action and beverage, food and antiallergic agent containing the strain - Google Patents

Abstract

The present invention provides a strain belonging to the genus Weisella, which has a stronger antiallergic action than previously known lactic acid strains, and contains beverages and foods containing the cells of this strain as active ingredients. It aims to provide antiallergic agents. The present invention provides a strain belonging to the genus Weissella and having an antiallergic action.

Description

  The present invention relates to a strain having an antiallergic action and beverages, foods and antiallergic agents containing the strain.

  Treatment of allergic diseases is generally pharmacotherapy using antihistamines, antiallergic agents, steroids and the like. Recently, however, lactic acid bacteria that act on intestinal immunity are attracting attention because they are effective in preventing and treating allergic diseases from the viewpoint of drug therapy and preventive medicine (Patent Documents 1 to 3). Intestinal immunity is an immune mechanism that eliminates pathogenic microorganisms orally taken in orally, and if it is possible to suppress intestinal immunity that reacts excessively, it is thought that it can contribute to the prevention and treatment of allergic diseases.

  For example, strains of the genus Bifidobacterium, infantitis, breve, longum and bifidum are effective in treating food allergies (Patent document 1), Enterococcus faecalis and Lactobacillus reuteri strains (patent document 2), and Lactobacillus paracacti and Lactobacillus p Streptococcus salivarius (Streptococcus sali varius) (patent document 3) has been reported to have an effect on bronchial asthma, allergic rhinitis, atopic dermatitis and the like.

JP-A-10-309178 JP 2000-95697 A JP 2005-139160 A

  However, pharmacological treatments such as antihistamines, antiallergic agents, and steroids act directly on molecules involved in the immune response in the blood and suppress the immune response necessary for maintaining the homeostasis of the living body. There is a point. In addition, in order to prevent chronic diseases such as allergic diseases, the activity of lactic acid bacteria reported so far has been insufficient in practice. Furthermore, until now, there has been no report that a strain of lactic acid bacteria belonging to the genus Weissella has an antiallergic effect.

  Accordingly, an object of the present invention is to provide a strain belonging to the genus Weissella, which has a stronger antiallergic action than lactic acid strains known so far. Another object of the present invention is to provide beverages and foods containing the cells of the strain belonging to the genus Weissella and antiallergic agents containing these as active ingredients.

  In order to achieve the above object, the present invention provides a strain belonging to the genus Weisella and having an antiallergic action.

  The present inventors have particularly effectively induced strains belonging to the genus Weisella to induce Th1-type cytokine production from mouse spleen cells and to suppress IgE production in spleen cells of ovalbumin (OVA) immunized mice. I found out. These effects are effective for the prevention and treatment of human allergic diseases, and their activities are significantly stronger than those of lactic acid bacteria strains reported so far. Lactic acid bacteria are bacteria that have been used for fermented foods for a long time, and are much safer for the human body than chemically synthesized antiallergic agents. As such a strain, Weissella confusa BP2865 (FERM BP-2865) is particularly preferable.

  The antiallergic action is preferably a production promoting action of interferon γ and / or interleukin 12.

  Interferon γ is a cytokine secreted by Th1 cells, which inhibits B cells from producing IgE and also has cellular immunity such as killer T cells and macrophages that attack viruses, filamentous fungi, and tuberculosis bacteria. Has an increasing effect. In addition, interleukin 12 is a cytokine secreted by antigen-presenting cells such as macrophages, has an action of inducing Th1 cells by stimulating NK cells, and also induces production of interferon γ by Th1 cells. Since the strain of the present invention can promote the production of interferon γ and / or interleukin 12 and suppress the production of IgE, it is possible to suppress the type I allergic reaction.

  The antiallergic action is preferably an IgE production inhibitory action.

  IgE is a causative substance that causes allergic diseases. That is, when allergen invades, IgE is produced in response to this, and IgE binds to mast cells and basophils to establish sensitization. Thereafter, when the same allergen enters, IgE recognizes the allergen, and inflammatory substances such as histamine are released from mast cells and basophils. This allergic reaction exhibits various symptoms such as bronchoconstriction and urticaria, and causes allergic diseases such as hay fever, allergic rhinitis, atopic dermatitis, and asthma depending on the site of onset. Since the strain of the present invention can suppress the production of IgE, it can suppress allergic reactions and can be used for the prevention and treatment of the above diseases.

  Moreover, this invention provides the drink and foodstuff containing the microbial cell of the said strain.

  The bacterial cells of the above strains have an antiallergic action and are safe for the human body, so that they can be used as a health food material in beverages and foods. Furthermore, since the said strain produces (gamma) -aminobutyric acid (GABA), it has an anti-stress effect | action, a blood pressure lowering effect | action, and a tranquilization effect | action, and its utilization value is high as a health food material.

  Moreover, this invention provides the antiallergic agent which contains the microbial cell of the said strain as an active ingredient.

  Since the bacterial cells of the above strain have an interleukin 12 and interferon γ production promoting action and an IgE production inhibitory action, if an antiallergic agent containing this bacterial cell as an active ingredient is produced, a chemically synthesized pharmaceutical product It can be used as an antiallergic agent with superior safety.

  ADVANTAGE OF THE INVENTION According to this invention, the strain which belongs to the genus Weissella which has stronger antiallergic action than the lactic acid strain known until now can be provided. Moreover, according to this invention, the drink, foodstuff, and antiallergic agent which contain the microbial cell of the said strain, are excellent in safety | security, and have an antiallergic action can be provided.

It shows the amount of interferon γ produced by the spleen cells of OVA immunized mice by the addition of OVA and the cell suspension of each strain. It shows the amount of interleukin 12 produced by the spleen cells of OVA immunized mice by the addition of OVA and the cell suspension of each strain. It shows the amount of interleukin 4 produced by the spleen cells of OVA immunized mice by the addition of OVA and the cell suspension of each strain. Interferon γ / interleukin 4 is calculated and graphed as an index of Th1 / Th2 balance of spleen cells of OVA immunized mice. It shows the effect of the cell suspension of each strain on the production of total IgE induced in the spleen cells of OVA-immunized mice by the addition of OVA. The effect of the cell suspension of each strain on the production of OVA-specific IgE induced in the spleen cells of OVA-immunized mice by the addition of OVA is shown. The amount of total IgE secreted into the peripheral blood of OVA immunized mice administered intraperitoneally with each strain is shown. It shows the production amount of OVA-specific IgE secreted into the peripheral blood of OVA immunized mice to which each strain was administered intraperitoneally.

  Hereinafter, preferred embodiments of the present invention will be described in detail.

  The strain of the present invention belongs to the genus Weissella and is characterized by having an antiallergic action.

  The genus Weissella includes the species Confusa, paramesenteroides, siberia, halotolerans, hellenica, kandelii, and kandeli. Kimchii species, Koreensis species, minor species, soli species, thairandensis species, viridescen species, but they are strains of Confucosa and Paramesenteroides species Preferably, it is a strain of Confucosa species.

  “Anti-allergic action” refers to the action of suppressing an allergic reaction. Here, allergy refers to a state in which an antibody is produced in a living body by ingestion or contact of a certain substance, and an excessive antigen-antibody reaction occurs due to reuptake or recontact of the same substance, thereby causing pathological symptoms. An allergic reaction refers to a phenomenon in which an immune reaction, which is a defense mechanism of a living body, attacks own cells that should not be excluded or food that has been ingested. The immune reaction involves antigen-presenting cells, T cells and B cells, and IgG and IgA are mainly produced as humoral immunity, whereas the allergic reaction mainly involves Th2 cells of T cells, IgE is made at concentrations 100 to 10,000 times higher than the normal immune response. This large amount of IgE binds to mast cells and promotes the release of inflammatory substances such as histamine and leukotriene in the mast cells.

  Anti-allergic effects include, for example, the action of acting on antigen-presenting cells, T cells or mast cells to suppress the production of IgE and the release of the inflammatory substances, or the action of shifting the Th1 / Th2 balance to the Th1 side. More specifically, IgE production inhibitory action, interferon γ and interleukin 12 production promotion action, and the like can be mentioned.

  The antiallergic action of the above strain is preferably an IgE production inhibitory action, an interferon γ production promoting action and / or an interleukin 12 production promoting action, more preferably two or more of these actions.

  Strains belonging to the genus Weisella can be easily separated from nature and can be identified by examining the base sequence of 16S ribosomal DNA. It can also be purchased from a cell bank such as ATCC.

  Among the strains belonging to the genus Weissella, the strains having antiallergic activity are, for example, i) interferon γ and interleukin 12 production promoting action on mouse spleen cells, and ii) spleen cells of ovalbumin (hereinafter referred to as OVA) immunized mice. It can be selected by testing the inhibitory effect of induced IgE production, iii) the inhibitory effect of IgE production secreted into the peripheral blood of OVA immunized mice, and the like.

  In i), spleen cells are isolated from the spleen of a mouse and cultured, and then a cell suspension prepared by sterilizing the cells of the test strain is added thereto and cultured for a certain period of time, which is secreted from the spleen cells What is necessary is just to measure the production amount of interferon (gamma) and interleukin 12 by ELISA etc., and to investigate the presence or absence of the production promoting effect of interferon (gamma) and interleukin 12.

  In ii), spleen cells of an OVA immunized mouse 2 weeks after the booster were isolated and cultured, and a cell suspension prepared by sterilizing the cells of the test strain and OVA were added thereto for a certain period of time. Culture may be performed, and the production amount of IgE secreted from spleen cells may be measured by ELISA or the like to examine the IgE production inhibitory action.

  In iii), a cell suspension prepared by sterilizing the cells of the test strain is administered into the abdominal cavity of an OVA mouse, reared for a certain period of time, and the amount of IgE secreted into the peripheral blood is determined by ELISA or the like. What is necessary is just to measure and to compare with the production amount of IgE secreted in the peripheral blood of the OVA mouse which is not administering the bacterial cell suspension intraperitoneally.

  Incidentally, Weissella confusa BP2865 (date of accession: April 13, 1990; accession number: FERM BP-2865), which belongs to the genus Weissella and has an antiallergic action, is an international deposit. Microorganism Industrial Technology Research Institute, Ministry of International Trade and Industry, Ministry of International Trade and Industry (currently, National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center; 1-6 Higashi 1-chome, Tsukuba City, Ibaraki, Japan )) And is available.

  The beverages, foods and antiallergic agents of the present invention are strains belonging to the genus Weissella and are characterized by containing the above strains having an antiallergic action.

  The bacterial cells of the strain can be added to beverages and foods for the purpose of preventing and treating allergic diseases. The said drink and food may consist only of microbial cells of said strain, and may contain the additive normally used in the said field | area. Examples of this additive include apple fiber, soybean fiber, meat extract, black vinegar extract, gelatin, corn starch, honey, animal and vegetable oils and fats, monosaccharides such as glucose, disaccharides such as sucrose, fructose and mannitol, dextrose and starch, etc. Polysaccharides, sugar alcohols such as erythritol, xylitol, and sorbitol, and vitamins such as vitamin C. These additives may be used alone or in combination.

  In addition, for the purpose of preventing allergic diseases and alleviating symptoms, food additives such as foods for specified health use, special nutritional foods, nutritional supplements, health foods, functional foods and sick foods You can also

  The anti-allergic agent of the present invention contains the cells of the above-mentioned strain as an active ingredient, and can be used as an anti-allergic agent with excellent safety if formulated with the addition of carriers, excipients and / or other additives it can. Examples of pharmaceutically acceptable additives include monosaccharides such as glucose, disaccharides such as sucrose, fructose and mannitol, polysaccharides such as dextrose and starch, sugar alcohols such as erythritol, xylitol and sorbitol, vitamin C Vitamins such as acacia gum, calcium phosphate, alginate, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose derivatives, tragacanth, gelatin, syrup, methyl hydroxybenzoate, talc, magnesium stearate, water, mineral oil These additives may be single species or plural species.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated concretely, this invention is not limited to these Examples.

1) About each strain used in the experiment Four strains (strains 1 to 4) belonging to Weicera confusa were isolated by the inventors and stored at 4 ° C. as lyophilized cells until used in the experiment. In addition, strain X belonging to Lactobacillus rhamnosus was isolated from commercially available yogurt and stored at 4 ° C. as lyophilized cells until used in experiments.

2) Evaluation of antiallergic action
i) Preparation of Cell Suspension In order to evaluate the antiallergic action of each of the above strains, a cell suspension of each strain was prepared. First, each strain was subjected to anaerobic conditions (gas composition of N ₂ : CO ₂ : H ₂ = 90: 5: 5), MRS liquid medium (manufactured by Difco, composition: 1% proteose peptone, 1% beef extract, 0.5% yeast extract, 2% glucose, 0.1% Tween 80, 0.5% ammonium citrate, 0.01% magnesium sulfate, 0.005% manganese sulfate, 0.2% dipotassium phosphate) After stationary culture for 3 days, the culture was centrifuged at 1,500 rpm for 10 minutes to recover the bacterial cells of each strain. The obtained microbial cells were washed with PBS, freeze-dried, and suspended in PBS to 1 mg / mL. The bacterial cell suspension thus obtained was autoclaved (121 ° C., 15 minutes) and used in the following experiments.

ii) Th1 cytokine production promoting action, Th2 cytokine production inhibiting action and IgE production inhibiting action of strains 1 to 4 on spleen cells of OVA immunized mice (in vitro)
A suspension of 4 strains belonging to the genus Weissella was added to the spleen cells of OVA immunized mice and cultured for a certain period of time. Interleukin 4) production inhibitory action and IgE production inhibitory action were evaluated. At that time, the action of the strain X belonging to Lactobacillus rhamnosus was also investigated and compared with the action of 4 strains belonging to the genus Weissera.

(Preparation of OVA immunized mice)
OVA immunized mice were prepared by intraperitoneally administering OVA to 6-week-old BALB / c mice (female) to artificially induce allergies. Specifically, first, 100 μg of OVA (Ovalbumin, Eggwhite, Purified; Worthington Biochemical Corporation) and 10 mg of aluminum hydroxide were dissolved in 1 mL of PBS to prepare an OVA antigen solution. The first immunization was performed after administration. One week later, an OVA antigen solution was prepared in the same manner as described above, and 200 μL of the solution was again administered into the abdominal cavity of the mouse for booster immunization. The allergic mice were boosted about 1 week later, and this mouse was used as an OVA immunized mouse in the following experiment.

(Preparation of spleen cells of OVA immunized mice)
First, the spleen was aseptically removed from an OVA immunized mouse 2 weeks after the booster immunization, and immersed in an RPMI1640 medium containing 10% FBS. Thereafter, the spleen was transferred to a petri dish and crushed with a pestle, and the suspension of mouse spleen cells was passed through a nylon filter net (Nippon Riken Instruments Co., Ltd.) having an opening of 70 μm and a wire diameter of 39 μm. The suspension of mouse spleen cells that passed through the nylon filter net was centrifuged at 1,500 rpm for 10 minutes, and the supernatant was discarded, followed by erythrocyte hemolysis reagent (0.16M ammonium chloride, Tris-HCl, pH 7.2). ) Was added to the precipitated mouse spleen cells and allowed to stand at room temperature for 5 minutes. Thereafter, fresh 10% FBS-containing RPMI1640 medium was added to wash the mouse spleen cells, centrifuged at 1,500 rpm for 10 minutes, and after discarding the supernatant, the number of cells was 5 × 10 ⁶ cells / mL. RPMI 1640 medium containing 10% FBS was added. The spleen cells of the OVA immunized mouse thus obtained were used for the following experiments.

(Measurement by ELISA of interferon γ, interleukin 12 and interleukin 4)
In order to measure interferon γ, interleukin 12 and interleukin 4, spleen cells of 2.5 × 10 ⁵ OVA immunized mice were seeded in a 96-well plate (cell density was 2.5 × 10 ⁶ cells / cell). mL), 37 ° C., 5% CO ₂ and cultured in 10% FBS-containing RPMI1640 medium. OVA (final concentration: 100 μg / mL) and cell suspension of each strain (final concentration: 1 μg / mL) were added to each well in which spleen cells of OVA-immunized mice were cultured, and cultured after 72 hours. The amount of each cytokine secreted into the supernatant was quantified by ELISA. At that time, a control in which PBS was added instead of the cell suspension was provided.

  The ELISA for quantifying interferon γ was performed as follows. First, 50 μL of a primary antibody (Rabbit anti-mouse / rat interferon-γ; BIO SOURCE) prepared to 1.25 μg / mL was added to each well of a 96-well plate (Maxisorp immunoplate; NUNC) at 4 ° C. The solid phase was allowed to stand overnight. Then, this 96-well plate was washed 3 times with Wash Buffer and blocked with 1% bovine serum albumin (BSA; Sigma). Subsequently, 50 μL of each culture supernatant or a preparation of interferon γ whose concentration was previously known was added to each well, reacted with the anti-interferon γ antibody as the primary antibody for 90 minutes, and washed 3 times with Wash Buffer. Subsequently, 50 μL of a secondary antibody (Anti-mouse / rat interferon-γ biotin conjugate; BIO SOURCE) prepared to 0.5 μg / mL was added to each well and allowed to react at room temperature for 90 minutes. Thereafter, each well was washed 5 times with Wash Buffer, streptavidin-HRP (BIO SOURCE) was added to react, washed 5 times with Wash Buffer, and then washed with TMB (tetramethylbenzidine) substrate solution (3, 3 ′, 5 , 5′-Tetramethylbenzidine (TMB) Liquid Substrate System (SIGMA)) was added and reacted. After sufficient color development, 50 μL of 2N sulfuric acid was added to each well to stop the reaction, and the absorbance at 450 nm was measured. A standard curve was prepared from the absorbance of the interferon γ preparation, and the amount of interferon γ produced by spleen cells of OVA mice was quantified from the standard curve.

  For interleukin-12 quantification, 1 μg / mL Purified anti-mouse IL-12 (p40 / p70) (BD Pharmingen) was used as the primary antibody, and 1 μg / mL Biotin anti-mouse IL-12 was used as the secondary antibody. Using (p40 / p70) (BD Pharmingen), ELISA was performed in the same procedure as the quantification of interferon γ. For the quantification of interleukin-4, 1 μg / mL Monoclonal Anti-mouse IL-4 Antibody (R & D Systems Inc.) was used for the primary antibody, and 1 μg / mL Biotinylated Anti-mouse IL-4AidDyDB (Systems Inc.) and ELISA was performed in the same procedure as the quantification of interferon γ.

  FIG. 1 shows the amount of interferon γ produced by the spleen cells of OVA-immunized mice by the addition of OVA and a cell suspension of each strain, FIG. 2 shows the amount of interleukin 12, and FIG. 3 shows the amount of interleukin 4. Is shown. FIG. 4 is a graph obtained by calculating interferon γ / interleukin 4 as an index of Th1 / Th2 balance. As a result, in the control to which OVA and PBS were added, production of interferon γ and interleukin 12 as Th1 type cytokines was hardly observed, and production of interleukin 4 as a Th2 type cytokine was remarkably induced. This suggested that the spleen cells of OVA-immunized mice had a Th1 / Th2 balance shifted to the Th2 side, and allergic reaction was enhanced.

  On the other hand, in the treatment group to which the cell suspension of OVA and strains 1 to 4 was added, the production of Th1-type cytokines interferon γ and interleukin 12 was promoted compared to the strain X belonging to control and Lactobacillus rhamnosus. Production of interleukin 4, which is a Th2-type cytokine, was remarkably suppressed. Moreover, the action of the bacterial cell suspensions of strains 1 to 4 significantly shifted the Th1 / Th2 balance of the spleen cells of the OVA immunized mouse to the Th1 side. From this result, it was suggested that the strains 1 to 4 have a strong antiallergic action.

(Measurement of total IgE and OVA-specific IgE by ELISA)
In order to measure total IgE, spleen cells of 2.5 × 10 ⁵ OVA-immunized mice were seeded in a 96-well plate (cell density was 2.5 × 10 ⁶ cells / mL), 37 ° C., 5% The cells were cultured in RPMI 1640 medium containing 10% FBS under CO ₂ conditions. Thereafter, OVA (final concentration: 100 μg / mL) and cell suspension of each strain (final concentration: 1 μg / mL) were added to each well in which spleen cells of OVA immunized mice were cultured, and 14 days had elapsed. Later, the amount of total IgE secreted into the culture supernatant was quantified by ELISA. For control, PBS was added instead of the bacterial cell suspension, and the amount was similarly determined by ELISA.

On the other hand, in order to measure OVA-specific IgE, a 48-well plate was seeded with 2.5 × 10 ⁶ spleen cells (cell density was 2.5 × 10 ⁶ cells / mL), 37 ° C., 5% The cells were cultured in RPMI 1640 medium containing 10% FBS under CO ₂ conditions. OVA (final concentration: 100 μg / mL) and cell suspension of each strain (final concentration: 1 μg / mL) were added to the spleen cells and cultured for 3 days. The spleen cells were washed three times with a new medium and OVA was performed. The bacterial suspension of each strain was added again to 1 μg / mL and cultured for 11 days, and IgE secreted in the culture supernatant was added to the OVA-specific IgE. As quantified by ELISA. For control, PBS was added instead of the bacterial cell suspension, and the amount was similarly determined by ELISA.

  ELISA for quantifying total IgE and OVA-specific IgE was performed as follows. First, 50 μL of anti-mouse IgE antibody (Mouse IgE ELISA Quantitation Kit; BETHYL Laboratories Inc.) prepared to 10 μg / mL was added to each well of a 96-well plate (Maxisorp immunoplate; NUNC) overnight at 4 ° C. It was solidified by placing. Then, this 96-well plate was washed 3 times with Wash Buffer and blocked with 1% bovine serum albumin (BSA; Sigma). Subsequently, each culture supernatant or a preparation of IgE whose concentration was previously known was added to each well, reacted with anti-mouse IgE antibody for 90 minutes, washed 3 times with Wash Buffer, and then biotinylated kit (Cygnus Technologies, Inc.). 50) of biotinylated OVA prepared in.) Was added to each well and allowed to react at room temperature for 90 minutes. Thereafter, each well was washed 5 times with Wash Buffer, streptavidin-HRP (BIO SOURCE) was added to react, washed 5 times with Wash Buffer, and then washed with TMB (tetramethylbenzidine) substrate solution (3, 3 ′, 5 , 5′-Tetramethylbenzidine (TMB) Liquid Substrate System (SIGMA)) was added and reacted. After sufficient color development, 50 μL of 2N sulfuric acid was added to each well to stop the reaction, and the absorbance at 450 nm was measured. A standard curve was prepared from the absorbance of the IgE preparation, and the amount of IgE produced by spleen cells of OVA mice was quantified from this standard curve. The amount of OVA-specific IgE was expressed as a relative value to the absorbance of the control (negative control) because there was no OVA-specific IgE preparation.

  FIG. 5 shows the effect of the cell suspension of each strain on the production of total IgE induced in the spleen cells of OVA-immunized mice by the addition of OVA. FIG. 6 shows the effect of the cell suspension of each strain on the production of OVA-specific IgE induced in the spleen cells of OVA-immunized mice by the addition of OVA.

  As a result, by adding the cell suspensions of strains 1 to 4, the total amount of IgE induced by OVA and the amount of OVA-specific IgE were remarkably suppressed. No. 1 was remarkable and almost 100% was suppressed.

  On the other hand, although strain X belonging to Lactobacillus rhamnosus suppressed the total amount of IgE induced by OVA and the production of OVA-specific IgE, it had a weaker effect than strain 1.

  From the above results, the strain 1 belonging to the genus Weissella has an interferon γ and interleukin 12 production promoting action and an IgE production inhibitory action, and exhibits a strong antiallergic action as compared with known lactic acid bacteria strains. Turned out to be. Strain 1 is named BP2865 and is an international depositary authority, Ministry of International Trade and Industry, Ministry of Industry and Technology, Institute of Microbial Technology (currently National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary Center; 1-chome East 1st, Tsukuba, Ibaraki, Japan The strain was deposited at the address No. 1 Chuo No. 6 (zip code 305-8666) (date of deposit: April 13, 1990; deposit number: FERM BP-2865).

iii) Inhibition of IgE production by BP2865 on OVA immunized mice (in vivo)
BP2865, which was found to have a strong antiallergic effect in in vitro experiments, was evaluated in vivo for its IgE production inhibitory effect on OVA immunized mice. At that time, the action of strain X belonging to Lactobacillus rhamnosus was also investigated and compared with the action of BP2865.

  OVA immunized mice were prepared by initial immunization and booster immunization with OVA (Ovalbumin, Eggwhite, Purified; Worthington Biochemical Corporation) as described above, and BP2865 or strain X bacterial suspension or PBS (control) Was administered intraperitoneally 200 μL once every two days from one week before the first immunization to one week after the booster immunization. Each bacterial cell suspension was prepared by suspending the lyophilized bacterial cells as described above in PBS to 1 mg / mL and autoclaving (121 ° C., 15 minutes).

  One week after booster immunization, blood was collected from the tail vein of mice, and the amount of total IgE and OVA-specific IgE in the serum separated therefrom was measured by ELISA. Measurement of total IgE and OVA-specific IgE in serum by LISA was performed in the same manner as described above.

  FIG. 7 shows the amount of total IgE secreted into the peripheral blood of OVA immunized mice to which each strain was administered intraperitoneally. FIG. 8 shows the production amount of OVA-specific IgE secreted into the peripheral blood of OVA immunized mice similarly administered intraperitoneally with each strain. * In the graph indicates that the control is statistically significant at p <0.05.

  As a result, the addition of the BP2865 cell suspension suppressed the total IgE amount secreted into the peripheral blood of the OVA mice by about 80% and the OVA-specific IgE by about 45%. The inhibitory effect of total IgE was statistically significant compared to the control administered with PBS.

  On the other hand, strain X belonging to Lactobacillus rhamnosus suppresses the production of total IgE and OVA-specific IgE secreted in the peripheral blood of OVA mice, but has a weaker action than BP2865, and is statistically different from control. It was not significant.

  From the above results, BP2865 belonging to the genus Weissella was found to be a strain having a strong antiallergic action.

3) Sensory test of fruit juice lactic acid fermentation broth manufactured using BP2865 A fruit juice lactic acid fermentation broth was manufactured using BP2865 having an antiallergic action, and sensory tests were performed on the aroma and flavor of the obtained fruit juice lactic acid fermentation broth.

  Table 1 shows the fruit type, sugar content and pH of the fruit juice used for lactic acid fermentation.

Each fruit juice was prepared by diluting grapefruit concentrated juice, apple concentrated fruit juice, white grape concentrated fruit juice and lemon concentrated fruit juice with sterilized water to have a predetermined sugar content, and the pH was adjusted by adding NaOH. Lactic acid fermentation was performed by inoculating 5.74 × 10 ⁸ cells of BP2865 into 100 mL of each fruit juice, stirring once a day, and standing at 30 ° C. for 72 hours under stationary conditions. After completion of fermentation, the turbidity of each lactic acid fermentation broth was measured with a spectrophotometer (Tytec Corp.), and the amount of lactic acid was measured with F-kit D- / L-lactic acid (JK International Corp.). The turbidity of each lactic acid fermentation broth was used as an index of BP2865 growth, and the amount of lactic acid was used as an index of the degree of lactic acid fermentation.

  Table 2 shows the turbidity, the amount of lactic acid, and the results of sensory test for each lactic acid fermentation broth.

  As a result, lactic acid fermentation of BP2865 progressed with all the fruit juices used in the test, and the fruit juice lactic acid fermentation broth was produced. In particular, in GF-10, AP-10, and LE-5, the flavor of the base fruit juice was maintained, and provided with a preferable property as a lactic acid fermented beverage.

  ADVANTAGE OF THE INVENTION According to this invention, the strain which belongs to the genus Weissella which has stronger antiallergic action than the lactic acid strain known until now can be provided. Moreover, according to this invention, the drink, foodstuff, and antiallergic agent which contain the microbial cell of the said strain, are excellent in safety | security, and have an antiallergic action can be provided.

Claims (7)

  A strain belonging to the genus Weissella and having an antiallergic action.   The strain according to claim 1, wherein the antiallergic action is an action of promoting production of interferon γ and / or interleukin 12.   The strain according to claim 1, wherein the antiallergic action is an IgE antibody production inhibitory action.   The strain according to any one of claims 1 to 3, which is Weissella confusa BP2865 (FERM BP-2865).   The drink containing the microbial cell of the strain according to any one of claims 1 to 4.   The foodstuff containing the microbial cell of the strain as described in any one of Claims 1-4.   The antiallergic agent which contains the microbial cell of the strain as described in any one of Claims 1-4 as an active ingredient.

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