KR100635354B1 - Composition for Prevention and Treatment of Food Allergy containing Bifidobacterium bifidum BGN4 as Active Ingredient - Google Patents

Abstract

The present invention relates to a composition for preventing and treating food allergy containing Bifidobacterium bifidum as an active ingredient and a method for preventing and treating food allergy by administering the composition. Bifidobacterium bifiderm BGN4, an active ingredient of the composition of the present invention, is excellent in preventing an immune response due to food allergy when administered before sensitizing food allergy.

Bifidobacterium Bifidem, Food Allergy

Description

Composition for Prevention and Treatment of Food Allergy containing Bifidobacterium bifidum BGN4 as Active Ingredient}             

Figure 1 is a schematic diagram showing the experimental schedule according to the sensitization using OVA and the administration time of the Bifidobacterium Bifidum BGN4 of the present invention.

2 is a graph of IgE level measurement confirming the effect of the Bifidobacterium bifiderm BGN4 pretreatment of the present invention on the allergic reaction by OVA sensitization.

Figure 3 is a graph showing the symptoms of the tail (A) and the symptoms (B) showing the symptoms of the allergic reaction in the mouse tail by Bifidobacterium bifiderm BGN4 pretreatment of the present invention.

Figure 4 is a graph of IgA level measurement present in the feces of mice confirmed the effect of the Bifidobacterium bifiderm BGN4 pretreatment of the present invention on the allergic reaction by OVA sensitization.

Figure 5 shows the OVA-specific IgG1 level (A), IgG2a level (B) and serum total IgG2a level of serum confirming the effect of the Bifidobacterium bifidum BGN4 pretreatment of the present invention on allergic reactions by OVA sensitization ( C), IgG1 level (D) graph.

6a shows IL-2 level (A), IFN-γ level (B), and IL-18 level of splenocytes confirmed that the Bifidobacterium bifiderm BGN4 pretreatment of the present invention affects allergic reactions by OVA sensitization. C) and IL-13 level (D), FIG. 6B is a graph of IL-5 level (A), IL-6 level (B), total IgG2a level (C) and total IgG1 level (D) of splenocytes.

The present invention relates to a composition for preventing and treating food allergy.

Food allergies are characterized by abnormal immune reactivity to food ingredients. Food allergies often begin in the first 1-2 years with the sensitization process, whereby the immune system responds to certain food proteins, especially allergen-specific immunoglobulin E (IgE). Once sensitized, individuals with allergies will experience the opposite effect by eating sufficient doses of allergic foods. Over time, most food allergies disappear even if allergy to some foods is long-lived. For example, most milk and egg allergies grow significantly but most peanut and walnut allergies do not appear (Wood RA. Pediatrics 2003; 111: 1631-7). The occurrence of food allergy peaks at 6-8% a year and gradually disappears until adolescence, after which the incidence is stabilized to 1-2% (Plaut M. J. Allergy Clin. Immunol . 1997; 100 : 7-10).

It is well known that newborns have a Th2-biased immune system (Martinez FD, Holt PG. Lancet 1999; 354: 12-5). It is thought to be converted to Th1 type by exposure to gastric bacteria and pathogens after birth (Gavett SH, et al., J Exp Med 1995; 182: 1527-36). ). Rautave et al. Proposed that the immune system develops by contact with food and microbial antigens in the immature gut of a newborn. The evolving innate gut microbiota has played a significant role in the development of the immune system, and the close interaction between the microbial community and host defense mechanisms balances the ability to tolerate non-toxic antigens and raise the inflammatory response to potential pathogens. And is essential for development (Rautava S, Isolauri E. Curr Issues Intest Microbiol 2002; 3: 15-22). The above microbial population is a major source of microbial stimulation in neonatal period. Thus, in the absence of childhood infection and generally in a clean environment, there is no Th2 to Th1 type conversion of T cell populations, so there is still a Th2 response, resulting in an increased allergy incidence (Hart AL, et al., Gut ecology. London: Martin Dunitz, 2002). Allergen inoculation of the postnatal immature immune system tends to convert newborns into potentially dangerous type 2 T-helper cell (Th2) -polarized memory (Prescott SL, et al., J Immunol 1998; 160: 4730 -7). The hygiene hypothesis is based on the assumption that this sensitization is much less induced in the facilitating presence of microbial population stimuli suitable to direct the immune response to the Th1 cytokine phenotype (Kirjavainen PV, et al., Allergy 1999; 54: 909-15).

Recently, many researchers have demonstrated that the administration of probiotics has an anti-inflammatory, resistant immune response, the absence of which leads to the development of atopic diseases. Because from a variety of probiotic, Bifidobacterium (Bifidobacterium) exists predominantly in Chapter newborn is considered to be the most influential. Bidodobacterium accounts for over 90% of bacterial populations in breastfed newborns, but the number decreases gradually over time (Bezkorovany A, Miller-Catchpole R. Biochemistry and physiology of Bifidobacteria, Boca Raton: CRC press, 1989).

Bifidobacteria include macrophage and lymphocytic activation, antibody activation (Hatcher GE, Lambrecht RS. J Dairy Sci 1993; 76: 2485-92), antibody production (Lee J, et al., Biosci Biotech Biochem 1993; 54: 2127-32; Link-Amster H, et al., FEMS Immunol Med Microbiol 1994; 10: 55-64) and proliferative responses in the spleen and Peyers patches (Hosono A, Lee J, et al. , Biosci Biothch Biochem 1997; 61: 312-6; Kado-Oka Y, Fujiwara S, Hirota T. Milchwissencshaft 1991; 46: 626-30).

Regarding allergies, He et al. Have shown that newborns with allergies have Bifidobacterium herds in adult Thais with high levels of B. adolesentis , while healthy newborns have high levels of BP. It has been reported to have a typical neonatal Bifidobacterium herd with B. bifidum and B. infants (He F, et al., International comference of Intestinal bacteriology 2002; 64). In addition, the adhesion of fecal bifidobacteria from healthy neonates was significantly higher in mucosal models of the adult intestine compared to newborns with allergies.

Hattori et al. Reported a significant improvement in allergic symptoms after 1 month of Bifidobacterium administration to infants with atopic dermatitis (Hattori K, et al., Arerugi . 2003; 52: 20-30). However, they did not reveal the relationship between microbial changes in feces and allergic symptoms. Therefore, there is a need to understand the effect of using Bifidobacteria depending on the timing.

It is well known that the timing of exposure to antigen or infection of newborns is of primary importance in the phenotype of Th1 cell responses (Holt PG, and Sly PD. J Exp Med 2002; 18: 1271-5; Culley FJ, et al. , J Exp Med 2002; 18: 1381-6). The function of Th1 is little or absent in newborns compared to Th2 (Garcia AM, et al., Immunol Res 2000; 22: 177-90; Admins B. Immunol Today 1999; 20: 330-5). Since Newborn's Th2 biased immune function is based on the immaturity of the immune system, probiotics can promote and easily develop their immune system. However, the Th2-biased allergic response in adults is clearly different from the newborn's immune system. The immune system of adult allergy sufferers is already mature. Thus, it is much more difficult to change T-cell populations in allergic responses in adults than in newborns. Thus, in a recent study, three-week-old mice with immature immune systems (just after the end of the gestational age) were used for OVA-induced allergy sensitization to easily alter Th1 cell populations. From the characteristics of the immune response with age, it was predicted that various effects could be made on allergic diseases depending on the timing of administration of probiotic Bifidobacterium. However, there have been no reports that the timing of administration of Bifidobacterium would be much more effective in treating or preventing allergic diseases.

Accordingly, the present inventors examined the immune response after administering Bifidobacterium to ovalbumin (OVA) -induced mice at various time points in order to verify the importance of administering probiotics Bifidobacterium to a food allergic reaction. As a result, the present invention was completed by confirming that the allergic suppression reaction when Bifidobacterium was administered before OVA induction was most effective.

Accordingly, it is an object of the present invention to provide a composition for preventing and treating food allergy containing Bifidobacterium bifidum as an active ingredient.

Another object of the present invention is to provide a method for preventing and treating food allergy by administering a composition containing the Bifidobacterium bipiderum as an active ingredient.

According to one aspect of the present invention, the present invention provides a composition for preventing and treating food allergy, which contains Bifidobacterium bifidum BGN4 as an active ingredient.

Bifidobacterium bifidom BGN4 (Professor Geun-Keok Ji, Department of Food and Nutrition, Seoul National University; Reference Journal of Food Protection, Vol. 63, (2000) 1133-1136 / International Journal of Food Microbiology Vol. 46 (1999) 231- 241.) is Gram-positive and has biphytobacterial properties such as F6PPK positive and club-like morphological characteristics. As a result of the 16S rRNA sequencing analysis, as shown in Table 1, Bifidobacterium bifiderm BGN4 was identified.

Abberviations: breve; B. breve , longum; B. longum , infantis; B. infantis, adol; B. adolescentis , ani; B. animalis, bifidum; B. bifidum , BGN-4; Bifidobacterium sp. BGN4.

According to a preferred embodiment of the present invention, Bifidobacterium bifidem BGN4, an active ingredient of the composition for preventing and treating food allergy, has the following effects on food allergy.

One. Oral Administration of Probiotic Bacteria Inhibits Allergic Reactions in Ovalbumin-Induced Allergic Mouse Models

In the present invention, to compare the action of probiotics and non-probiotic bacteria on allergic suppression, B. bifidum BGN4, Lactobacillus cassia ( L. bifidum BGN4 ) in an obalbumin-induced allergic mouse model. casei) 911 and E. coli The effect of MC4100 was tested. All bacterial treatments reduce IgE levels in mouse serum. Allergic symptoms in the tails of mice are also reduced by the treatment of bacteria. The IgA levels in the stool samples of the Bifidobacterium and Lactobacillus treated groups were lower compared to the sensitized group, but the E. coli treated group showed no difference. In splenocyte culture supernatants, the treatment of bacteria increases the level of Th1 type cytokines but decreases the level of Th2 type cytokines. The Bifidobacterium and Lactobacillus treated groups maintained their normal body weight compared to the untreated group, but the body weight of the E. coli treated group remained lower than the untreated group during the experimental period.

These results indicate that oral administration of Bifidobacterium or lactobacterium can suppress IgE-induced ovalbumin oversensitization and maintain normal growth of ovalbumin-sensitized mice. Although the immune response is partially suppressed, general allergic symptoms can also be suppressed by administration of E. coli.

2. The timing of administration of Bifidobacterium affects the development of ovalbumin allergy in mice.

Previous studies have shown that treatment with Bifidobacterium has an inhibitory effect on allergic reactions. However, the various anti-allergic actions of many Bifidobacterium have not yet been identified. Therefore, the present inventors observed the change in immune response by administering the Bifidobacterium of the present invention two weeks before or two weeks after the sensitization of ovalbumin in order to determine the effect of the point of administration of Bifidobacterium. The ovalbumin specific IgE level in the group sensitized with ovalbumin continuously increased for 6 weeks, whereas the pretreated group of bifidobacterium of the present invention stopped increasing the IgE level. However, during the experimental period, the IgE level of the pretreatment group was as low as that of the no treatment group.

The Bifidobacterium pretreatment group and the posttreatment group of the present invention have reduced symptom severity of the tail of the mouse. The ovalbumin specific IgA level in the post-treatment group is no different from the sensitized group, but the ovalbumin specific IgA level in the pretreatment group is lower than the sensitized group. Pretreatment with Bifidobacterium of the present invention reduces IgG1 and IgG2a levels in serum, but not in the post-treatment group. In splenocyte culture supernatants, the levels of IL-2 and IFN-γ are increased by Bifidobacterium pretreatment and post-treatment of the present invention, while IL-5, IL-13 and IL-18 levels are inhibited. These results show that the Bifidobacterium pretreatment of the present invention inhibits the occurrence of allergy. The Bifidobacterium post-treatment of the present invention alters the cellular response in splenocytes but partially changes the overall response. Therefore, it can be seen that administration of the Bifidobacterium of the present invention before sensitization or before allergic development is much more effective in suppressing allergy. The allergic inhibitory reaction in the post-treatment of Bifidobacterium of the present invention is much weaker than pretreatment. Therefore, in order to alleviate allergic diseases, it is much more effective to take the Bifidobacterium of the present invention before the allergic disease is induced.

3. Viability and intactness of the Bifidobacterium of the present invention affect allergic suppression

In order to verify the effective use of the Bifidobacterium of the present invention and its immuno-modulatory action to suppress allergic reactions, the present inventors heat killed and crushed Bifidobacterium in the ovalbumin-induced mouse model. Administered. Both viable and heat killing bacteria of Bifidobacterium inhibit allergic reactions in serum levels of IgE and IgG1, symptoms in the tail and antibodies of the splenocyte supernatant. Shredding fungi reduce ovalbumin-specific IgE levels, but do not reduce IgG1 and IgG2a levels in serum, nor change IFN-γ levels in splenocyte culture supernatants. The results of the present invention can be seen that there is no difference in the anti-allergic action between live bacteria and heat killing bacteria.

Administration of surviving bacteria is expected to alter the microbial population in the gastrointestinal tract. However, bacteria tend to die by competition from gastric acid, pancreatic enzymes or other bacteria in the stomach. It is not known whether the survival of bacteria is indicative of an in vivo immune system difference. The duration of Bifidobacterium administration is not sufficient to compare the effects of viable or dead cells. Therefore, long-term experiments can help to understand the effects of antiallergic reactions.

However, the inhibitory effect in allergic reactions of Bifidobacterium crushed bacteria was much weaker in the severity of symptoms in the tail and cytokine patterns in the splenocyte culture supernatant. However, allergic reactions in serum IgE still inhibited. Although bacterial components that may affect allergies have not been fully understood, physical disruption is expected to destroy the effective components of Bifidobacteria. In addition, they may lose their active ingredients while passing through the host's digestive organs or by processes in immune cells.

In conclusion, the Bifidobacterium of the present invention can alleviate cellular and humoral allergic reactions in ovalbumin induced mice by oral administration. Heat kill bacteria are also effective in suppressing allergic reactions. The inhibitory effect of the present invention Bifidobacterium on allergy is caused by intact cellular components, so that the whole cells of Bifidobacterium are much more effective than crushed bacterium.

As described in detail above, the Bifidobacterium bifiderm BGN4 of the present invention is effective in suppressing an immune response to food allergy. Preferred food allergens for the prophylaxis or treatment by administering the composition of the present invention may be selected from the group consisting of ovalbumin, peanut protein, fish, nuts such as walnuts and chestnuts, shellfish such as shrimp and crabs, soybeans, wheat and the like. And, more preferably, ovalbumin.

In addition, the Bifidobacterium bifiderm BGN4 of the present invention is preferably a live bacterium itself or a heat killed bacterium. According to a preferred embodiment of the present invention, live or heat killing bacteria of Bifidobacterium bifidum BGN4 showed similar food allergic reaction inhibitory effect, but the physically crushed bacteria were shown to decrease the effect.

According to another aspect of the present invention, the present invention provides a method for preventing and treating food allergy by administering a composition containing Bifidobacterium bifidum BGN4 as an active ingredient.

In the method of the present invention, the characteristics of Bifidobacterium bifiderm BGN4, which is an active ingredient of a composition administered to prevent and treat food allergy, are the same as those described in the composition, and thus the description thereof is to be avoided to avoid excessive complexity of the specification. Omit.

According to a preferred embodiment of the present invention, the Bifidobacterium Bifidum BGN4 included in the composition is preferably administered before ingesting a food allergen, and is administered at least two weeks before the ingestion of the food allergen and continuously administered. More preferred.

In addition, the Bifidobacterium bifiderm BGN4 of the present invention is preferably administered at (10 ⁸ )-(10 ⁹ ) cfu per day relative to the total body weight to be treated.

Since the method of the present invention is effective in preventing and treating food allergies, the treatment target is not particularly limited since it is applied to all animals capable of causing food allergies.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are only for illustrating the present invention, and the content of the present invention is not limited by the following examples.

<Example>

Experimental Materials and Methods

mouse

Three-week-old female mice weighing 11-13 g were purchased from SLC (Hamamatsu, Japan) and maintained in an OVA-free diet. Mice were raised in plastic cages and watered freely. Temperature and humidity were adjusted to 23 ± 1 ° C. and 55 ± 10%. Mice were reared in a chamber controlled in an animal environment with a 12:12 hour light: dark cycle. Animal-related experiments were conducted according to management guidelines of Seoul National University. Mice were sensitized at 5 weeks of age.

microbe

B. bifidum BGN4, Lactobacillus, from feces of healthy people, according to previously known literature (Ji GE, Lee SK, Kim IH.Kor J Food Sci Technol 1994; 26: 526-31) L. casei 911 and E. coli MC4100 was isolated. Bifidobacterium and Lactobacillus were anaerobic for 24 hours at 37 ° C. in Lactobacilli-MRS broth (Difco, Detroit, MI, USA) containing 0.05% L-cysteine (Sigma, St Louis, MO, USA). Incubated. To prepare the mouse diet, bacterial cells were centrifuged at 4,000 g for 40 minutes at 4 ° C. using a centrifuge (Hanil, Seoul, Korea), and then washed twice with sterile phosphate buffered saline. The pellets were then vacuum dried (Ilshin, Seoul, Korea) and mixed with the diet of mice. Heated and killed bacterial cells were used to stimulate spleen cells for use in vitro testing.

Sensitization of gastric internal antigen and preparation of diet

Before oral immunization, mice were starved for 2 hours. Immunization was performed by administering 10 g of cholera toxin (CT) intragastrically (ig) at days 0, 1, 2, 7, 21 and 35 using a stainless steel blunt feeding needle. OVA (Sigma, St Louis, MO, USA) was used as antigen and CT and Concanavalin A (Con A) were purchased from Sigma (St Louis, MO, USA). Each mouse was fed with 0.2 ml of phosphate-buffered saline solution (PBS, pH 7.2) containing OVA and CT. Mice in the untreated group (Group 1) treated with PBS without OVA and CT were used as negative controls. Five groups of mice were used for this experiment (FIG. 1). Mice in groups 2-4 showed OVA-sensitized allergic reactions and mice in groups 3-4 were treated with different bacterial powders. Mice in group 2 received OVA and CT but no bacteria as sham treatment.

Bifidobacterium treated mice were dieted with lyophilized 0.2% Bifidobacterium bifidem BGN4 diet pellets. Mice were fed with bacterial powder for two weeks, starting two weeks before the first sensitization or two weeks after the first sensitization. Serum was obtained from tail blood vessels and stored at -80 ° C to verify the antibody response of the serum.

Measurement of Serum OVA-Specific IgE, IgG1 and IgG2a, Total IgE, IgG1 and IgG2a, Spleen IFN-γ, IL-2, IL-5, IL-13, IL-18, IgG1 and IgG2a Levels

After the initial sensitization, blood was collected from the tail blood vessel every week. Serum was separated from blood and stored at -80 ° C. OVA-specific-IgE, IgG1 and IgG2a levels were measured by ELISA. Specifically, Nunc-Immuno-Maxisorp plates (Nunc, Roskilde, Denmark) were coated with coating buffer pH 9.6 (Sigma, St Louis, Mo, USA) containing 5 μg / ml OVA and left overnight at 4 ° C. I was. Plates were blocked and washed. Each sample was added to the plate and allowed to react overnight at 4 ° C. The plate was washed and then treated with biotinylated mouse anti-mouse IgE, IgG1 or IgG2a monoclonal antibody (2 μg / ml) to detect OVA-specific-IgE, IgG1 and IgG2a for 1 hour at room temperature. Reaction at The reaction was developed at room temperature for 30 minutes using 3,3 ', 5,5'-TMB (tetramethylbenzidine) substrate (Fluka, Neu-Ulm, Switzerland). The color reaction was stopped using 6N H ₂ SO ₄ and measured at 450 nm. Equivalence of IgE, IgG1 or IgG2a was calculated by comparison with reference curves prepared using mouse whole IgE, IgG1 or IgG2a standards, respectively.

Total IgE, IgGl and IgG2a from serum were detected by ELISA. Plates were coated with 2 μg / ml of mouse monoclonal anti-mouse IgE, IgG1 or IgG2a, respectively, to detect the antibodies. 100 μl of biotinylated rat monoclonal anti-mouse IgE, IgG1 or IgG2a was added after the step dilution of serum was added. As mentioned above, HRP-conjugated streptavidin was used for detection and plates were developed with TMB substrate. IFN-γ, IL-2, IL-5, IL-13 and IL-18 levels from the supernatant of spleen cultures were detected by ELISA in the same manner as described above. All antibodies used in the experiments of the present invention were purchased from Pharmingen (San Diego, CA, U.S.A).

Measurement of OVA-specific IgA and Total Fecal IgA

Extraction from stool pellets was performed by the method disclosed by Marinaro et al. (Marinaro M., et al., J Exp Med 1997; 185: 415-27). Specifically, 100 mg of pellet was mixed with 1 ml of PBS containing 0.1% NaN ₃ and reacted at 4 ° C. for 2 hours. The pellet was then vortexed for 10 minutes. After centrifugation (3000 rpm, 20 minutes), the supernatant was separated and stored at -70 ° C. For analysis, plates were coated with a coating buffer containing 5 μg / ml OVA. After washing and blocking, 100 μl of fecal extract was added to each well and allowed to react overnight at 4 ° C. After washing the plate, biotinylated murine anti-mouse IgA monoglonal antibody (2 μg / ml) was added to the plate and further reacted at room temperature for 1 hour. After washing, avidin-peroxidase was added and reacted at room temperature for 1 hour. The reaction was developed with TMB (Fluka, Neu-Ulm, Switzerland) at room temperature for 30 minutes. The color reaction was stopped using 6N H ₂ SO ₄ and measured at 450 nm. Equivalence of IgA was calculated by comparison with a reference curve created using the whole mouse IgA standard.

To measure total IgA, plates were coated with a coating buffer containing murine anti-mouse IgA capture antibody (μg / ml). The plate was then blocked and washed in the same manner as described above. Stool extract (1:50 dilution) was added to the plate and reacted overnight at 4 ° C. After washing the plate, 100 μl of biotinylated murine anti-mouse IgA was added to each well. Additional steps thereafter were performed in the same manner as described above. IgA levels were calculated by comparison with reference curves made using the whole mouse IgA standard.

Preparation of Spleen Cells

Spleens were isolated from mice in each group (n = 6). At 7 weeks, the spleen cells were prepared by killing the mice and gently separating the spleens with a glass slide and passing them through a 200 gauge stainless mesh. Erythrocytes in splenocytes were lysed with 0.83% NH ₄ Cl-tris buffer (pH 7.6). Cells were cultured in RPMI 1640 culture medium (Gibco BRL, NY, USA) containing 10% fetal bovine serum (Gibco BRL) and 1% penicillin / streptomycin (Gibco BRL). The cultured cells were stimulated with Con A (10 μg / ml), lyophilized BGN4 powder (100 μg / ml) and OVA (1 mg / ml) for 48 hours in a 24-well flat bottomed plate and then 10% Cultured under 5% CO ₂ conditions in a RPMI 1640 culture medium (Gibco BRL) containing fetal serum (Gibco BRL) and 1% penicillin / streptomycin (Gibco BRL) until a concentration of 5 × 10 ⁶ cells / ml was reached. . The supernatant was separated to predict cytokine production. All cell cultures were incubated in a 5% CO ₂ (Sanyo, Japan) incubator at 37 ° C. with humidity.

Measurement of Oversensitization Response

Allergy symptoms were verified using the following scoring system after sacrifice: 0, no symptoms; 1, puffiness of the tail; 2, with 1-2 scabs in the tail; 3, with 3-4 scabies in the tail; 4, with 5-6 scabies in the tail; 5, more than 7 scabies in the tail. Scoring of symptoms was performed in a blind manner; The scores were scored by 10 well trained graduate students in determining identity between samples.

Statistical analysis

All data are expressed as mean standard standard error of mean (SEM), which is indicated directly in the figure. Data was analyzed using the SAS (Release 8.01, O, USA) program. For immunoglobulin and cytokine levels, differences between groups were analyzed by ANOVA test followed by Duncans complex range test for complex comparison. p Values <0.05 was determined to be significant.

Experiment result

1. Effect of Bifidobacterium Bifidum BGN4 on IgE Production

In order to monitor the effect of Bifidobacterium bifidem BGN4, serum was isolated from each group every two weeks after OVA-sensitization. OVA-specific IgE in serum of each group is shown in FIG. 2. The level of OVA-specific IgE in the sensitized group (abbreviated as "sham group") gradually increased up to 6 weeks after the initial sensitization and was significantly different from that in the untreated group at 6 weeks. Statistically different (untreated group, 4310 ± 263 ng / ml; Siamese group, 9887 ± 175 ng / ml; pretreated group, 4805 ± 245 ng / ml; post-treated group, 5723 ± 207 ng / ml). However, OVA-specific IgE levels in the post-treatment group were lower than the same group at 6 weeks, but did not differ significantly from the untreated group at 6 weeks. The pretreated group did not differ from the untreated group in OVA-specific IgE levels.

2. Symptoms of the tail

Mice sensitized with OVA and CT showed some damage to the tail during the experiment. Immediately after administration of OVA and CT, mice in the Siamese group began to scratch their tails and had injuries including bleeding (FIG. 3). The score of symptoms in the tail of the Siamese group was the highest and significantly different from the untreated group. No damage or scratching was observed in the pretreated group and the symptom severity was the highest among the experimental groups. The scores of symptoms in the tail of the post-treated group were significantly lower than in the Siamese group but higher than in the untreated group.

In addition, the results of the heat kill bacteriophage showed low scores compared to the live bacterium and the untreated group, and the result of the crushed bacterium showed higher scores than the untreated or live bacterium.

3. OVA-specific mucosal IgA

OVA-specific mucosal IgA levels in fecal extracts isolated at 6 weeks were measured. The IgA of the post-treated and pretreated groups and the Siamese group showed no significant difference from the OVA-specific IgA in the untreated group (FIG. 4). (Untreated group, 24.6 ± 5.6 μg / ml; Siamese group, 36.2 ± 6.4 μg / ml; pretreatment group, 48.1 ± 10.2 μg / ml; post-treated group, 54.6 ± 8.5 μg / ml).

In addition, the IgA level of the heat killing bacteria pretreatment group and the crushing killing bacteria pretreatment group also showed the same result as the pretreatment level of the live bacteria.

4. Levels of IgG1 and IgG2a in Serum

OVA-specific IgG1 levels in the serum of the co-treated group at 6 weeks were significantly higher than the untreated group (FIG. 5), while the OVA-specific IgG1 levels in the pre-treated group were similar to the untreated group. OVA-specific IgG1 levels in the post-treatment group were higher than the untreated and pre-treated groups. In contrast, OVA-specific IgG2a levels of the Siamese group were not different from the untreated group. IgG2a levels in the post-treatment group were significantly higher than in the pretreatment group. There was no difference in total IgG1 and IgG2a between all groups, and OVA-specific IgG1 levels in the sterile treatment group were higher than the untreated group.

5. Antibody and Cytokine Levels in Spleen Cell Culture

To determine if treatment of bacteria affected the systemic Th1 / Th2 response to BGN4, ConA and OVA-promoting, the levels of cytokines in the spleen cell culture supernatants were measured (FIG. 6). IFN-γ levels in the pretreatment group or the posttreatment group were higher than the Siamese group. In addition, the level of IL-2 in the BGN4 treated group was higher than the Siamese group.

IL-18 levels in the Siamese group were much higher than in the pretreatment and posttreatment groups. IL-13 and IL-5 levels in the pretreatment and posttreatment groups were lower than the Siamese group.

In contrast, the total IgG1 level in the spleen of the Siamese group was higher than the pretreatment or posttreatment group. In addition, the total IgG2a level in the spleen of the Siamese group was higher than the pretreatment or posttreatment group.

6. Effect of Treatment of Bifidobacterium Bifi4 BGN4 on Body Weight

Initial mean weights between groups did not differ significantly. The mean weight of the pretreatment group at 2 weeks after initial sensitization was significantly higher than that of the other groups, but there was no difference between the other groups at 2 weeks (Table 2). However, at 7 weeks, the final weight of the Siamese group was significantly lower than that of the untreated group. The final weight of the pretreatment group was not different from the untreated group, but the final weight of the post-treatment group was much lower than that of the untreated group. The pretreatment group maintained high levels during the experiment.

group 2 weeks 3 weeks 4 Weeks 5 Weeks 6 Weeks Week 7 No treatment 17.2 ± 0.6 ^b 18.8 ± 0.5 ^a 19.1 ± 0.8 ^a 22.4 ± 0.6 ^a 22.9 ± 0.7 ^a 23.3 ± 1.1 ^a Concurrent processing 18.8 ± 0.4 ^b 18.5 ± 0.3 ^ab 19.9 ± 0.3 ^a 20.5 ± 0.5 ^b 21.2 ± 0.4 ^b 21.0 ± 0.6 ^b Pretreatment 19.5 ± 0.3 ^a 19.7 ± 0.4 ^a 20.9 ± 0.3 ^a 21.2 ± 0.2 ^ab 21.8 ± 0.4 ^ab 22.9 ± 0.4 ^a After treatment 16.9 ± 0.3 ^b 19.0 ± 0.2 ^a 20.6 ± 0.4 ^a 21.5 ± 0.3 ^ab 21.2 ± 0.5 ^b 21.9 ± 0.1 ^ab

1) Experimental Diet:

Untreated group: 0.2% corn starch

Concurrent Treatment Group: 0.2% Corn Starch

Pretreatment group: 0.2% Bifidobacterium Bifidum BGN4 administered 2 weeks prior to initial sensitization

Post-treatment group: 0.2% Bifidobacterium Bifidum BGN4 administered 2 weeks after initial sensitization

Each value represents the mean ± SEM of 6 values for each group.

Superscript notation indicates significant differences from Duncan's composite test ( p <0.05).

As described above, Bifidobacterium bifidum BGN4, an active ingredient of the composition of the present invention, has an excellent effect of preventing an immune response caused by food allergy when administered before sensitizing food allergy, thereby alleviating allergy symptoms. It can be usefully used.

Claims (6)

Bifidobacterium bifidum ( Bifidobacterium bifidum ) A composition for preventing and treating food allergy containing BGN4 as an active ingredient. According to claim 1, wherein the Bifidobacterium bifidum ( Bifidobacterium bifidum ) BGN4 is a composition for preventing and treating food allergy, characterized in that the live or heat kill bacteria. The food allergy according to claim 1, wherein the allergen causing food allergy is selected from the group consisting of ovalbumin, peanut protein, fish, nuts such as walnuts and chestnuts, shellfish of shrimp and crabs, soybeans and wheat. Prophylactic and therapeutic compositions. delete delete delete

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