HK1081432B - Antiallergic agent, utilization thereof in the manufacture of a medicament for reducing allergy - Google Patents

Description

Antiallergic agent and pharmaceutical use thereof

Technical Field

The present invention relates to an antiallergic agent. The invention also relates to the use of anti-allergic agents for attenuating allergy, and to methods of attenuating allergy.

Background

In many countries such as japan, the number of allergic patients increases year by year, and it is said that one of every three adults is an allergic disease patient in japan. Allergic diseases are divided into four major classes, i.e. types I to IV, according to the mechanism of their action. Allergic rhinitis, such as certain types of pollinosis, bronchial asthma, and atopic dermatitis are known as immunoglobulin E (IgE) -dependent type I allergy, and an increase in antigen-specific IgE antibodies in the blood increases the risk of inducing allergic symptoms.

The mechanism for initiating the type I allergy is as follows. The invasion of antigens such as pollen, house dust or mites into the body produces IgE antibodies specific to these antigens and binds to the fce receptors on mast cells or on the surface of basophils to sensitize the subject. Thereafter, the antigen binds to the IgE antibody once invaded into the body to form a complex, thereby inducing degranulation, and histamine in the particles and chemical mediators such as the mouth イコトリエン are released, thereby showing allergic reaction symptoms.

At present, antagonists of chemical mediators, mainly represented by antihistamines, and steroid agents used as anti-inflammatory agents are used for the treatment of allergic diseases. However, both agents provide only symptomatic treatment and steroids suppress the overall immune response and are therefore associated with side effects. In addition, agents that inhibit the release of chemical mediators by inhibiting degranulation have also been used, but there has not been a fundamental therapeutic agent that specifically reduces the main factor causing the onset of disease, i.e., IgE antibodies.

Further, when it is necessary to administer an antiallergic agent for a long period of time, an antiallergic agent which is easy to take and highly safe is desired. Therefore, there is a need for new anti-allergic agents with these properties.

Summary of The Invention

The object of the present invention is to provide an antiallergic agent which can reduce the amount of IgE antibodies involved in the induction of type I allergy, improve allergic constitution, can be administered easily and is highly safe, and a method for attenuating allergy.

To solve the above problems, the present inventors constructed a mouse model in which antigen-specific IgE levels were significantly increased, but IgG levels were not greatly increased. In this model, the suppression of IgE levels by various strains of lactic acid bacteria that may affect the intestinal immune system was investigated, and it was found that some of the various lactic acid bacteria tested had particularly good inhibitory effects on the production of IgE, thereby completing the present invention.

According to the present invention, there is provided an antiallergic agent comprising as an active ingredient a lactic acid bacterium selected from the group consisting of lactic acid bacteria belonging to Lactobacillus acidophilus (Lactobacillus acidophilus), lactic acid bacteria belonging to Lactobacillus fermentum (Lactobacillus fermentum), and combinations thereof.

The invention also provides that said lactic acid bacteria are antiallergic agents selected from the group consisting of the Lactobacillus acidophilus strain CL0062 (deposited in the depositary center under the number FERM BP-4980), CL92 (deposited in the depositary center under the number FERM BP-4981), belonging to the species Lactobacillus acidophilus, and combinations thereof.

The invention also provides an antiallergic agent wherein the lactic acid bacterium is Lactobacillus fermentum strain CP34 (deposited in the center of the International patent organism under the accession number FERM BP-8383) belonging to Lactobacillus fermentum.

The present invention also provides an antiallergic agent which is characterized in that the amount of antigen-specific IgE antibodies in blood is increased by continuing antigen stimulation of the nose in a mouse rhinitis model, and the amount of antigen-specific IgE antibodies in blood can be decreased by orally administering the antiallergic agent of the present invention.

The invention also provides the application of the specific lactic acid bacteria in preparing the medicine for weakening the allergic reaction.

The present invention further provides a method of attenuating an allergic response comprising administering to a subject in need of such attenuation an effective dose of the anti-allergic agent.

Brief Description of Drawings

FIG. 1 is a graph showing the change in immunoglobulin levels in the blood of high IgE mice in example 1.

FIG. 2 is a graph showing the results of experiments in example 2 in which administration of fermented milk to high IgE mice inhibited the OVA-IgE levels.

FIG. 3 is a graph showing the results of experiments in example 3 in which administration of fermented milk to high IgE mice inhibited the OVA-IgE levels.

FIG. 4 is a graph showing the results of experiments in example 4 in which administration of fermented milk to high IgE mice inhibited the OVA-IgE levels.

FIG. 5 is a graph showing the results of experiments in which fermented milk was administered to humans to inhibit OVA-IgE levels in example 5.

FIG. 6 is a graph showing the results of experiments in which fermented milk was administered to humans to inhibit OVA-IgE levels in example 5.

Embodiments of the invention

The antiallergic agent of the present invention comprises, as an effective ingredient, a lactic acid bacterium selected from the group consisting of Lactobacillus acidophilus and Lactobacillus fermentum belonging to the lactic acid bacterium group, and a combination thereof.

The lactobacillus belonging to Lactobacillus acidophilus is preferably Lactobacillus acidophilus strain CL0062 (deposited at the central agency of franchised biologies (central agency of 5-1-1, No. 6 at 3/4 th of Kyowa, Japan) under the accession number FERM BP-4980), Lactobacillus acidophilus strain CL92 (deposited at the central agency of franchised biologies under the accession number FERM BP-4981 at 3/4 th of 1994), or a combination thereof. Further, Lactobacillus fermentum strain CP34 (deposited in the International patent organism depositary, No. FERM BP-8383, 5/23, 2002) is preferably used as the Lactobacillus fermentum. These three strains have been deposited according to the budapest treaty on the international recognition of the deposit of microorganisms for patent procedures. Once the patent rights were granted, the restrictions on public access to Lactobacillus fermentum strain CP34 were removed altogether. In addition, the Lactobacillus acidophilus strains CL0062 and Lactobacillus acidophilus CL92 are now publicly available.

The lactobacillus acidophilus strain CL0062 has the following bacteriological properties:

(morphological characteristics)

1) Cell shape: the bacillus is a bacillus which is capable of producing,

2) presence or absence of motility: the number of the Chinese characters is zero,

3) existence of spores: the number of the Chinese characters is zero,

4) gram staining: the protein is positive and the protein is negative,

(physiological Properties)

1) Catalase: the negative result is negative, and the negative result is negative,

2) indole production: the negative result is negative, and the negative result is negative,

3) nitrate reduction: the negative result is negative, and the negative result is negative,

4) demand for oxygen: the anaerobic biological fertilizer is a mixture of a facultative anaerobic fertilizer,

5) growth at 15 ℃: the number of the Chinese characters is zero,

6) DL-lactic acid is formed from glucose by homolactic fermentation without gas formation,

7) whether or not to form acids from sugars

Glucose: + melibiose: +

Lactose: + raffinose: +

Mannose: + mannitol: -

Fructose: + sorbitol: -

Galactose: + esculin: +

Sucrose: + salicin: +

Arabinose: -N-acetylglucosamine: +

Maltose: + amygdalin: +

Xylose: -gentiobiose: +

Rhamnose: -melezitose: -

Cellobiose: + dextrin: +

Trehalose: + starch: -

The lactobacillus acidophilus strain CL92 has the following bacteriological properties:

(morphological characteristics)

1) Cell shape: the bacillus is a bacillus which is capable of producing,

2) mobility: the number of the Chinese characters is zero,

3) spore formation: the number of the Chinese characters is zero,

4) gram staining: the protein is positive and the protein is negative,

(physiological Properties)

1) Catalase: the negative result is negative, and the negative result is negative,

2) production of indole: the negative result is negative, and the negative result is negative,

3) nitrate reduction: the negative result is negative, and the negative result is negative,

4) demand for oxygen: the anaerobic biological fertilizer is a mixture of a facultative anaerobic fertilizer,

5) growth at 15 ℃: the number of the Chinese characters is zero,

6) DL-lactic acid is formed from glucose by homolactic fermentation without gas formation,

7) whether or not to form acids from sugars

Glucose: + melibiose: -

Lactose: + raffinose: +

Mannose: + mannitol: -

Fructose: + sorbitol: -

Galactose: + esculin: +

Sucrose: + salicin: +

Arabinose: -N-acetylglucosamine: +

Maltose: + amygdalin: +

Xylose: -gentiobiose: +

Rhamnose: -melezitose: -

Cellobiose: + dextrin: -

Trehalose: + starch: -

Lactobacillus fermentum strain CP34 has the following bacteriological properties:

(morphological characteristics)

1) Cell shape: the bacillus is a bacillus which is capable of producing,

2) mobility: the number of the Chinese characters is zero,

3) spore formation: the number of the Chinese characters is zero,

4) gram staining: the protein is positive and the protein is negative,

(physiological Properties)

1) Catalase: the negative result is negative, and the negative result is negative,

2) demand for oxygen: the anaerobic biological fertilizer is a mixture of a facultative anaerobic fertilizer,

3) DL-lactic acid is formed from glucose, and gas (+),

4) formation of acids from sugars

Arabinose: -cellobiose: \ u

Xylose: -lactose: +

Melibiose: -trehalose: \ u

Rhamnose: -amygdalin: \ u

Ribose: + raffinose: \ u

Glucose: + melezitose: -

Mannose: -mannitol:

fructose: + sorbitol: -

Sucrose: + esculin: -

Maltose: + salicin: -

In the antiallergic agent of the present invention, the content of the lactic acid bacteria is not particularly limited, and may be suitably adjusted depending on the convenience of production, the preferable daily dose, etc., and for example, 1X 10 is preferable for a liquid formulation⁷Cells/ml to 1X 10¹⁰Cells/ml.

The antiallergic agent of the present invention may contain other components in addition to the lactic acid bacteria. Examples of such other components may include additives such as excipients, and the components of the culture medium discussed later.

The antiallergic agent of the present invention can be prepared by culturing lactic acid bacteria in a medium.

As the culture medium, any medium can be used as long as the lactic acid bacteria can grow therein, and synthetic media such as animal milk, skim milk, whey, MRS medium, GAM medium, BL medium, Briggs liver broth medium, and the like can be used. The temperature for the cultivation may be 25 ℃ to 50 ℃, preferably 35 ℃ to 42 ℃. The culture time may be 3 hours to 48 hours, preferably 8 hours to 12 hours. The culture medium after the culture or the medium treated as necessary can be used as the antiallergic agent of the present invention. For example, cells harvested from a cultured medium by centrifugation or filtration, lyophilized products thereof, heat-treated cells or ground cells can be used as the antiallergic agent of the present invention. Moreover, the microbial cells in the above form can be further formulated or incorporated into various food materials such as beverages, tablets, pastes or breads, and used as the antiallergic agent of the present invention.

The method of administering the antiallergic agent of the present invention is not particularly limited, but oral administration is preferred. For example, when the composition is orally administered to a human daily, the number of cells in the composition may be 2X 10⁹One or more, preferably 2X 10¹⁰These doses may be administered once a day or multiple times a day.

As shown in the examples below, the antiallergic agent of the present invention effectively suppresses the amount of IgE antibodies, and on the other hand, it is considered to be highly safe because it contains edible fungi as an active ingredient.

The method of attenuating allergy of the present invention comprises the step of administering to a subject in need of such attenuation an effective amount of an anti-allergic agent as described above. The subject can be an animal, such as a human or other mammal.

The antiallergic agent of the present invention can effectively inhibit the amount of IgE antibodies in the living body, and is easy to take and highly safe. Thus, the agent is useful for inhibiting allergic reactions involving excessive amounts of IgE antibodies.

Examples

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

Example 1

(preparation of high IgE mice)

Male BALB/c mice were purchased from Charles River, Japan and were bred under conditions in which CE-2 (manufactured by CLEA, Japan) was taken as a feed ad libitum. Mu.g of ovalbumin (hereinafter abbreviated as OVA, produced by SIGMA CHEMICAL CO.) and 2mg of aluminum hydroxide (WAKO PURE CHEMICALINDUSTRIES, LTD.) as an adjuvant were suspended in 300. mu.l of physiological saline. The first immunization was performed by intraperitoneal injection of this suspension into 10 or more mice aged six weeks on the first and fourth days of sensitization. For the second sensitization, the nose of each mouse was immersed in a 25mg/ml OVA antigen solution formed by dissolving in physiological saline for three seconds, and this immersion operation was repeated three times as one cycle. High IgE mice were prepared by daily immersion twice daily from day 10 to 16.

Blood was partially collected from the fundus vein of the high IgE mouse on the 1 st and 17 th days of sensitization, and serum samples were obtained from the collected blood. Serum samples were measured for OVA-specific IgE (hereinafter abbreviated as OVA-IgE), total IgE, and total IgG according to the methods described below. The results are shown in FIGS. 1(a) to 1 (c).

From the results shown in FIGS. 1(a) to 1(c), it is understood that the increase in total IgE and OVA-IgE in blood by sensitization is significantly larger than that of IgG. Thus, a mouse allergy model in which the overall immune function was not changed and the amount of IgE in blood and antigen-specific IgE were increased was constructed.

(measurement of blood OVA-IgE)

The assay was performed by sandwich ELISA. Mu.l of a physiological saline solution (trade name: AAM11, manufactured by Dainippon pharmaceutical Co., Ltd.) containing 10. mu.g/ml of goat anti-mouse IgE polyclonal antibody was added to each well of a 96-well immunoplate (manufactured by CORNING INCORPORATED), and incubated overnight at 4 ℃. The plate was washed with phosphate buffer (containing 137mM NaCl, 2.7mM KCl, 8.1mM Na)₂HPO₄And 1.5mMKH₂PO₄Hereinafter abbreviated as PBS) was washed three times, each well was filled with PBS containing 0.5% casein and incubated at room temperature for 3 hours, and then washed three times with PBS. 100. mu.l of a serum sample diluted 1/10 with PBS was added to each well and reacted at 4 ℃ overnight. After washing the plate four times with PBS, 100. mu.l of 0.5% -casein-PBS containing 10. mu.g/ml of biotinylated OVA (biotin-labeled OVA) labeled with the Biotinylation kit (produced by AMERICAN QUALEX INTERNATIONAL INC.) was added to each well and reacted at room temperature for 2 hours. After the plate was washed five times with PBS, 100. mu.l of a PBS solution containing 1. mu.g/ml of streptavidin-peroxidase (manufactured by SIGMA CHEMICAL CO.) and 0.5% casein was added to each well, and reacted at room temperature for 1 hour. After the plate was washed five times with PBS containing 0.1% Tween20, 100. mu.l of 0.2M citric acid buffer (prepared by mixing 0.2M citric acid and 0.2M sodium citrate and adjusting pH to 5) containing 600. mu.g/ml of 2, 2' -azino-bis- (3-ethylbenzthiazoline-6-sulfonic acid) (hereinafter abbreviated as ABTS, produced by BOEHRINGER MANNHEIM) and 0.006% hydrogen peroxide was added to each well, and color development was performed at 37 ℃ for 3 hours under masking. After completion of the reaction, OD was measured₄₀₅And OD₄₉₂By OD₄₀₅value-OD₄₉₂The value obtains a true color rendering value.

In one aspect, blood samples are taken from mice that have been injected intraperitoneally five times (once a week) with 25mg/ml ova in saline. Serum prepared from the blood sample was used as standard serum. This standard serum was 1/10 diluted with PBS and then stepwise diluted two-fold with non-immunized serum to prepare a dilution for plotting a detection curve. The resulting dilutions were subjected to color value measurement according to the above method to obtain detection curves. Based on this detection curve, the amount of OVA-IgE in the serum sample was obtained, and the relative amount was determined with the amount of OVA-IgE in the standard serum as 1.

(measurement of Total IgE in blood)

Mu.l of a physiological saline solution containing 10. mu.g/ml of goat anti-mouse IgE polyclonal antibody (trade name AAM11, manufactured by Dainippon pharmaceutical Co., Ltd.) was added to each well of a 96-well immunoplate (manufactured by CORNING INCORPORATED), and incubated overnight at 4 ℃. Plates were washed three times with PBS, wells were filled with PBS containing 0.5% casein, incubated at room temperature for 3 hours, and plates were washed three times with PBS. Then 50. mu.l of 1/25-diluted serum sample in PBS containing 0.5% casein was added to each well and reacted at 4 ℃ overnight. After the plate was washed four times with PBS, 50. mu.l of a PBS solution containing 2. mu.g/ml of a biotin-labeled anti-mouse IgE antibody (produced by YAMASA CORPORATION) and 0.5% casein was added to each well, and reacted at room temperature for 2 hours. After the plate was washed five times with PBS containing 0.1% Tween20, 50. mu.l of a PBS solution containing 1. mu.g/ml of streptavidin-peroxidase and 0.5% casein was added to each well, and reacted at room temperature for 1 hour. After the plate was washed five times with PBS containing 0.1% Tween20, 50. mu.l of 0.2M citric acid buffer (pH5) containing 300. mu.g/ml ABTS and 0.006% hydrogen peroxide was added to each well, and the reaction was masked at room temperature for 20 to 30 minutes. Subsequent measurement of OD₄₀₅。

On the other hand, mouse anti-DNP-IgE (manufactured by YAMASA CORPORATION) replacement serum samples were dissolved in PBS containing 0.5% casein at various concentrations, and the same procedure as above was performed to obtain detection curves. Based on this detection curve, the total IgE amount in the serum sample was calculated.

(measurement of Total IgG in blood)

Mu.l of a physiological saline solution containing 1. mu.g/ml of goat anti-mouse IgG (H + L) antibody (trade name 62-6500, ZYMED LABORATORIES, INC.) was added to each well of a 96-well immunoplate (manufactured by CORNING INCORPORATED), and incubated overnight at 4 ℃. Plates were washed three times with PBS and each well was filled withPBS containing 0.5% casein and incubated for 3 hours at room temperature, and the plates were washed three times with PBS. To each well 50 μ l of serum samples diluted with 5% casein in PBS1/1000 were added and reacted overnight at 4 ℃ and the plates were washed four times with PBS. To each well, 50. mu.l of a PBS solution containing peroxidase-labeled anti-mouse IgG (. gamma.) antibody (CAPPEL LABORATORIES, manufactured by INC.) at 2. mu.g/ml and 0.5% casein was added and reacted at room temperature for 2 hours. After the plate was washed five times with PBS containing 0.1% Tween20, 50. mu.l of 0.2M citric acid buffer (pH5) containing 300. mu.g/ml ABTS and 0.006% hydrogen peroxide was added to each well, and the reaction was masked at room temperature for 20 to 30 minutes. Subsequent measurement of OD₄₀₅。

On the other hand, instead of serum samples, purified mouse IgG (produced by CAPPEL LABORATORIES) was dissolved in PBS containing 0.5% casein at various concentrations, and the same operation as above was carried out to obtain detection curves. Based on this detection curve, the total IgG amount in the serum sample was calculated.

Example 2

(comparison of Effect of various lactic acid bacteria)

Each of the lactic acid bacterial strains shown in Table 1 was pre-cultured in MRS medium at 37 ℃ overnight, and the cells were collected by centrifugation at 3000 rpm for 10 minutes. The collected cells were fermented at 37 ℃ by adding 9% (W/V) reduced de-esterified milk containing 0.1% (W/V) yeast extract (produced by DIFCO) until the milk coagulated. After fermentation, the total number of bacteria was measured for each fermented milk. The results are shown in Table 1.

TABLE 1

Bacterial strains	Total bacteria count (cells/ml)
		Lactobacillus acidophilus (Lactobacillus acidophilus) CL92(BP-4981)	1.9×10
Lactobacillus bulgaricus (Lactobacillus bulgaricus) CP1812	1.5×10
		Lactobacillus fermentum (Lactobacillus fermentum) CP34	5.3×10
Lactobacillus helveticus (Lactobacillus helveticus) CP790	2.4×10
		Lactobacillus johnsonii (Lactobacillus johnsonii) CP2551	2.7×10
Lactobacillus plantarum (Lactobacillus plantarum) CP2172	5.9×10
		Lactobacillus rhamnosus ATCC53103	1.0×10

Subsequently, hyper-IgE mice were prepared in the same manner as in example 1, and OVA-IgE in blood on day 18 of sensitization was measured in the same manner as in example 1. The mice were divided into groups of 10 mice each, and the amount of OVA-IgE in the blood of each group was equalized. From day 19 to day 21 of sensitization, each of the fermented milks described above, unfermented 9% (W/V) reduced de-esterified milk or unfermented 9% (W/V) reduced de-esterified milk containing 750 μ g of cyclophosphamide was forcibly administered to each group of mice at a dose of 1ml per day through a gastric tube for three days. On day 22 of sensitization, a blood sample of the mouse was collected from the fundus vein, and a serum sample was prepared. Blood levels of OVA-IgE and total IgG were measured. As a control, a blood sample of a mouse which had been sensitized in the same manner but had not been given fermented milk or the like was collected in the same manner, and blood OVA-IgE and total IgG amounts were measured. The results are shown in FIG. 2.

As shown in FIG. 2, a significant suppressive effect (p < 0.01) on the amount of OVA-IgE was observed in the group of mice given Lactobacillus acidophilus or Lactobacillus fermentum fermented milk compared to the group given unfermented skim milk. No significant difference was observed in total IgG levels in blood (not shown).

Example 3

The procedure of example 2 was followed except that the lactic acid bacteria strains shown in Table 2 were used. The results of the total bacteria count measurement in each fermented milk are shown in Table 2. The results of the blood OVA-IgE assay are shown in FIG. 3.

TABLE 2

Bacterial strains	Total bacteria count (cells/ml)
		Lactobacillus acidophilus (Lactobacillus acidophilus) CL0062(BP-4980)	4.40×10
Lactobacillus gasseri CP2209	4.30×10
		Lactobacillus reuteri ATTCC23272	9.60×10
Bifidobacterium breve (Lactobacillus) CP2425	1.30×10

As shown in FIG. 3, a significant suppressive effect (p < 0.01) on the amount of OVA-IgE was observed in the group of mice given Lactobacillus acidophilus fermented milk compared to the group given unfermented skim milk. No significant difference was observed in total IgG levels in blood (not shown).

Example 4

(comparison of the Effect of lower doses)

Lactobacillus acidophilus strain CL92 and Lactobacillus fermentum strain CP34 were pre-cultured overnight at 37 ℃ in MRS medium, respectively, and the cells were collected by centrifugation at 3000 rpm for 10 minutes. The collected cells were cultured overnight at 37 ℃ in MRS-supplemented medium, and the cells were collected by centrifugation at 3000 rpm for 10 minutes. The cell number of each strain was measured, and the cells were treated with 9% skim milk at 1X 10⁶The cells/ml concentration were suspended to obtain a suspension.

Subsequently, hyper IgE mice were prepared in the same manner as in example 1, and blood OVA-IgE at day 18 of sensitization was measured in the same manner as in example 1. The mice were divided into groups of 10 mice each, and the amount of blood OVA-IgE was equalized among the groups. From day 19 to day 21 of sensitization, the above suspension was forcibly administered to each group of mice at a daily dose of 1ml for three days through a gastric tube. On day 22 of sensitization, a blood sample of the mouse was collected from the fundus vein, and a serum sample was prepared. The amount of OVA-IgE and total IgG in the blood was measured. The results are shown in FIG. 4.

As shown in fig. 4, a significant suppressive effect (p < 0.01) on the amount of OVA-IgE was observed in the group of mice administered with lactobacillus acidophilus strain CL92 and lactobacillus fermentum strain CP34, compared to the group administered with unfermented skim milk. No significant difference was observed in total IgG levels in blood (not shown).

The reduction rate d of OVA-IgE when each suspension was administered was obtained by the formula d ═ 1- (b/a), where a represents the standard rate of OVA-IgE when fed unfermented skim milk and b represents the standard rate of OVA-IgE when fed each suspension. The number of cells in suspension x (cells/ml) necessary for a half reduction of OVA-IgE in this experimental system was obtained by the formula x ═ (s × 0.5)/d, where s (cells/ml) represents the bacterial concentration of the suspension administered to mice, and is assumed to be proportional to the reduction rate d. Using this formula, the number of cells x of each of the strains used in examples 2 and 3 was obtained. The results are shown in table 3.

TABLE 3

Bacterial strains	Number of cells required (cells/ml)
		Lactobacillus acidophilus CL92(BP-4981)	1.0×10
Lactobacillus bulgaricus CP1812	2.0×10
		Lactobacillus fermentum CP34	1.4×10
Lactobacillus helveticus CP790	3.3×10
		Lactobacillus johnsonii CP2551	3.5×10
Lactobacillus plantarum CP2172	7.0×10
		Lactobacillus rhamnosus ATCC53103	2.9×10
Lactobacillus acidophilus CL0062(BP-4980)	5.0×10
		Lactobacillus gasseri CP2209	3.1×10
Lactobacillus reuteri ATTCC23272	3.3×10
		Bifidobacterium breve CP2425	1.1×10

Example 5

(clinical Effect for human)

13 subjects suffering from perennial allergic rhinitis (average age 22.9. + -. 6.1 years, 6 women and 7 men) were given an observation period of 2 weeks, after which time Lactobacillus acidophilus strain CL 928.0X 10 was administered⁸To 1.3X 10⁹Cells/ml fermented milk 100 ml/day for 4 weeks. During which a questionnaire for subjective symptoms was issued and symptoms were scored on the basis of responses according to the "severity classification of allergic rhinitis" provided by the japanese society of allergy. The symptoms of rhinitis were diagnosed at intervals according to the Japanese society of allergy index. Further, blood was collected at regular intervals, and the IgE antibody titer in blood was measured. Also, the lowest temperature of the day was recorded during the test. The severity of nasal congestion, the frequency of nose blowing and the minimum temperature of the subjects during the test are shown in figures 5 and 6.

During the test period, the daily minimum temperature fluctuated greatly, decreasing by more than 10 ℃ from 14 ℃ on day 1 of ingestion (11 months and 15 days) to 3.7 ℃ on the last 1 day of ingestion (12 months and 13 days). Even under such conditions that tend to exacerbate the symptoms of rhinitis, nasal congestion shows a tendency to improve two weeks after the start of ingestion (Wilcoxon test: p < 0.1), and a significant improvement is observed four weeks after the start (Wilcoxon test: p < 0.05). The frequency of blowing shows a tendency to decline three weeks after the beginning of ingestion (Wilcoxon test: p < 0.1). During ingestion, a downward trend in sneezing frequency, a reduction in lower turbinate swelling, and a reduction in total IgE antibody titers in the blood were observed.

Claims (4)

1. An antiallergic agent comprising Lactobacillus acidophilus strain CL92 deposited in the trusted center for franchises under accession number FERM BP-4981 as an effective ingredient.

2. Use of the Lactobacillus acidophilus strain CL92 deposited in the registered biosignal center under the accession number FERM BP-4981 for the preparation of a medicament for attenuating allergic reactions.

3. The use of claim 2, wherein the allergic reaction is an IgE-dependent type I allergy.

4. The use of claim 2, wherein the allergy is allergic rhinitis.