CN116407569A - Antiallergic probiotic composition and application thereof - Google Patents
Antiallergic probiotic composition and application thereof Download PDFInfo
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- CN116407569A CN116407569A CN202111659393.3A CN202111659393A CN116407569A CN 116407569 A CN116407569 A CN 116407569A CN 202111659393 A CN202111659393 A CN 202111659393A CN 116407569 A CN116407569 A CN 116407569A
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- lactobacillus
- allergens
- probiotic composition
- bifidobacterium
- allergen
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Abstract
The invention provides the use of a probiotic composition comprising bifidobacteria, lactobacilli, cocci and bacilli in a ratio of viable bacteria count of (50-150): 5-15): 0.5-1.5 in the manufacture of a medicament for the prevention and/or treatment of diseases caused by allergic reactions. The invention also provides a pharmaceutical composition for preventing and/or treating diseases caused by anaphylactic reaction and a kit containing the pharmaceutical composition, which comprises a probiotic composition and an allergen preparation such as mite pathogen inhibitor and the like. The probiotic composition can remarkably treat anaphylactic reaction, effectively resist external allergen invasion and reduce the probability of allergic diseases of organisms. Meanwhile, the probiotic microecological preparation contained in the composition has harmless ingredients and no side effect on the body.
Description
Technical Field
The invention belongs to the field of medicines, and relates to an antiallergic probiotic composition, application thereof in preparing medicines for preventing and/or treating diseases caused by anaphylaxis, a pharmaceutical composition containing the probiotic composition and an allergen preparation, and a kit containing the pharmaceutical composition.
Background
Currently, allergic diseases have become a common public health problem. In 2005 epidemiological investigation of allergic diseases in 30 countries by the World Allergic Organization (WAO) showed that 22% of the total population in these countries had allergic diseases such as allergic rhinitis, asthma, conjunctivitis, eczema, food allergy, drug allergy, etc. Moreover, the incidence of allergic diseases is increasing year by year, creating a huge medical burden. How to effectively prevent and treat allergic diseases has become a current research focus.
Allergy refers to an inappropriate immune response of the body to substances which are otherwise harmless in the environment, belongs to systemic diseases, and has the following characteristics: multiple children, recurrent episodes, multiple allergens, multiple organ involvement. Allergic diseases occur at a high rate in industrialized developed countries, one of the reasons being that miniaturization of the family scale and reduced incidence of childhood infection reduce the chances of contact of microorganisms which play an important role in the process of host immune maturation in infants. Common allergens pollen, house dust mites, moulds, pharmaceuticals, foods and animal hair and dander. The most common allergic diseases are rhinitis, asthma and atopic dermatitis. Side reactions caused by currently common allergic reactions include: edema, increased vascular permeability, airway obstruction, localized congestion, nasal mucosa edema, nasal mucosa irritation, bronchoconstriction, increased mucus secretion, airway inflammation, vomiting, diarrhea, itching, urticaria, rash.
Allergic asthma is a chronic inflammatory disease. Symptomatic treatment of allergic diseases is achieved by improving allergic symptoms through the use of antihistamines, beta-agonists and corticosteroids. Many asthmatics whose symptoms are difficult to control are frequently administered orally with glucocorticoids, which cause a number of adverse effects. Children in many countries in the world have about 10% of the probability of suffering from asthma, which is caused by various factors, and the disease is easy to repeatedly attack, so that the physical health and normal learning of the children are seriously affected. At present, the infants suffering from this kind of diseases are often treated clinically by inhaling glucocorticoids and long-acting beta receptor agonists, but the effect is still not ideal, and a need exists for searching for more effective treatment schemes. The immunotherapy regimen for allergic diseases includes: subcutaneous immunotherapy, such as U.S. TO-204 dust mite extract preparation, denmark An Tuo Data-house dust mite allergen preparation, arroger-mite allergen injection in Germany, for use in subcutaneous immunotherapy in the United states, japan; sublingual immunotherapy, such as chang di-dust mite drops, odaCtra house dust mite inhibitors in the united states; oral immunotherapeutic means, such as ragweed, birch pollen preparation; intranasal local specific immunotherapy; intralymphatic immunotherapy, etc.
Mites are one of the most common allergens for children in China, and can cause allergic rhinitis and asthma. Treatment of allergic rhinitis, asthma includes avoiding contact with allergens, drug management, AIT, patient education, etc. Treatment with inhaled glucocorticoid-based drugs can rapidly and effectively alleviate symptoms, but does not alter the natural course of allergic rhinitis and asthma. Specific immunotherapy (specific immunotherapy, SIT) is currently the only causal treatment that can affect the natural course of allergy, but there are still reports of adverse effects of this treatment. For example, the local response is manifested as local redness and itching around the injection site; systemic reactions are manifested by symptoms associated with asthma (cough, wheezing, chest distress, decreased peak flow rate of exhalation), upper respiratory symptoms (itching throat), vomiting, abdominal pain, etc. The mite allergen preparation is prepared from inactivated mite pure culture, such as mite body, insect body fragments, mite excrement, larva, ovum, etc., and can be used for in vivo diagnosis or desensitization treatment of allergic diseases caused by mite allergen. Currently, commercial dust mite allergen preparations Andes contain preparations of mite allergen extracts, which are indicated for the desensitization treatment of patients with mild-moderate allergic asthma and/or allergic rhinitis with history of house dust mite sensitization. The treatment is carried out in two stages, namely an initial treatment stage and a maintenance treatment stage, wherein the initial treatment stage is 1-15 weeks, the maintenance treatment stage is 17-51 weeks, the treatment period is too long, the clinical application is not facilitated, and inconvenience is brought to patients. In addition, the use of dust mite allergen preparation for safe and effective treatment may cause adverse reactions such as local anaphylactic reaction, systemic anaphylactic reaction, anaphylactic shock, etc., for example, local anaphylactic reaction is mainly manifested by local swelling, redness and itching at the injection site of the medicine; systemic allergic reactions manifest mildly as hay fever symptoms and moderately as urticaria or asthma; anaphylactic shock is manifested by dyspnea, systemic urticaria, angioedema, laryngeal edema accompanied by wheezing, asthma, hypotension, nausea, vomiting, diarrhea, abdominal pain, loss of consciousness, convulsion, coma, etc. No drug capable of reducing adverse reactions while treating antiallergic reactions has been reported in the prior art.
Lactobacillus belongs to the genus of the accepted safety (Generally Regarded as Safe, g.r.a.s), which is currently known to be a group of microorganisms living in the body beneficial to host health, and can play a role in maintaining human health and regulating immune function. Specific strains of lactobacillus have been found to colonize the intestinal mucosa and assist in maintaining human and animal health, typically at live bacteria levels of over 10 billion in concentration. In recent years, various probiotics living bacteria preparations are developed at home and abroad, the basic guiding idea is that strains of normal physiological flora of human beings or animals are screened and cultured to prepare living bacteria preparations in various dosage forms through various ways, and then the living bacteria preparations are returned to the original environment in a throwing mode, so that the natural physiological effect of the living bacteria preparations is exerted. The research of probiotics for health-care foods and medicines in China is started later, and bifidobacteria are used for various health-care foods only in the 90 th century. The probiotic medicine formulations appearing on the market at present are roughly divided into three types, namely capsules, such as Mei Chang An, poly-g, intestinal metaplasia, bei Fei Dai, lizhu intestinal music and the like; granules and powders, such as mammy, pefeikang, chang Lekang, etc.; tablets, such as pro-bacteria, jin Shuangqi, probiotic compositions, lactic acid bacteria, etc., which are mainly used for intestinal related digestive diseases.
For some allergens, no medicine for treating corresponding allergic diseases has been developed at present, and the most main treatment mode is to use medicines for controlling allergic symptoms (i.e. symptomatic medication); however, the medicine cannot change the progress of allergic diseases, so that the curative effect cannot be achieved fundamentally.
Disclosure of Invention
The invention aims to provide a probiotic composition which is safe to use and has an antiallergic effect, and is used for resisting external allergen invasion and reducing the probability of allergic diseases of organisms. Allergen-induced specific IgE secreted by plasma cells is a prerequisite for type I hypersensitivity and is also an important evaluation index of the level of hypersensitivity. IgE is combined with receptors on the surfaces of basophils or mast cells to release hypersensitive response factors such as histamine, cytokines, chemokines, lipid mediators and the like to cause hypersensitive response, and the hypersensitive response state of animals is measured by detecting the IgE level in the bodies of test animals through immune detection means such as ELISA and the like. IL-2 is a cytokine produced by T cells and acting mainly on T cells, and has the main functions of promoting proliferation and differentiation of T cells, promoting synthesis of cytokines and promoting regulation of development of T cells; can lead to proliferation and activation of NK cells, proliferation of B cells and antibody synthesis; the ability to stimulate activation of cytotoxic lymphocytes and macrophages is an important factor in mediating and maintaining the immune protective function of the body, but is not directly involved in type I hypersensitivity. We have also studied the body level of IL-2 to investigate the effect of drugs on the protective immune function of the body while treating allergic symptoms. Through a great deal of experimental screening and research, the inventor of the invention surprisingly discovers that the existing four-linked live bacteria preparation mainly used for treating digestive system diseases such as diarrhea, constipation, dyspepsia and the like caused by dysbacteriosis of intestinal tracts has remarkable antiallergic activity, can be used for preventing and treating allergic diseases, and does not have side effect on human bodies by adopting the active probiotics without harmful ingredients. In addition, the inventor of the invention also unexpectedly discovers that the combination of the four-combined live bacteria preparation and the allergen preparation can change the treatment process of allergic diseases, realize better treatment of allergic reaction and radically achieve cure effect; in addition, the adoption of the treatment means can greatly shorten the antiallergic treatment period of the tetrad live bacteria preparation or the allergen preparation; meanwhile, the composition can obviously reduce adverse reaction in the process of treating anaphylactic reaction, and provides great favorable conditions for antiallergic treatment.
In one aspect, the present invention provides the use of a probiotic composition in the manufacture of a medicament for the prevention and/or treatment of a disease caused by an allergic reaction, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli.
According to an embodiment of the use of the present invention, the probiotic composition may comprise bifidobacteria, lactobacilli, cocci and bacilli in a ratio of the number of viable bacteria of (50-150): (50-150): (5-15): (0.5-1.5).
According to a preferred embodiment of the use according to the invention, the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2).
According to a preferred embodiment of the use according to the invention, the probiotic composition comprises a total number of viable bacteria not lower than 1 x 10 5 cfu/g. More preferably, the viable count of bifidobacteria in the probiotic composition is at least 1 x 10 6 cfu/g; the viable count of Lactobacillus is at least 1×10 6 cfu/g; the number of living bacteria of the coccus is at least 1×10 6 cfu/g; and the viable count of the bacillus is at least 1 multiplied by 10 5 cfu/g. According to an embodiment of the use according to the invention, the bifidobacteria may be selected from one or more of bifidobacterium adolescentis (Bifidobacterium adolescentis), bifidobacterium animalis (Bifidobacterium animalis), bifidobacterium bifidum (Bifidobacterium bifidum), bifidobacterium breve (Bifidobacterium breve), bifidobacterium infantis (Bifidobacterium infantis) or bifidobacterium longum (Bifidobacterium longum). According to a preferred embodiment of the use according to the invention, the bifidobacteria are bifidobacteria infantis.
According to an embodiment of the use according to the invention, the lactobacillus may be selected from one or more of lactobacillus acidophilus (Lactobacillus acidophilus), lactobacillus casei (Lactobacillus casei), lactobacillus crispatus (Lactobacillus crispatus), lactobacillus delbrueckii subspecies bulgaricus (bulgaricus) (Lactobacillus delbrueckii subsp. Bulgarisus) (Lactobacillus bulgaricus)), lactobacillus delbrueckii subsp. Lactis (Lactobacillus delbrueckii subsp. Lactis), lactobacillus fermentum (Lactobacillus fermentium), lactobacillus grignard (Lactobacillus gasseri), lactobacillus helveticus (Lactobacillus helveticus), lactobacillus johnsonii (Lactobacillus johnsonii), lactobacillus paracasei (Lactobacillus paracasei), lactobacillus plantarum (Lactobacillus plantarum), lactobacillus reuteri (Lactobacillus reuteri), lactobacillus rhamnosus (Lactobacillus rhamnosus), lactobacillus salivarius (Lactobacillus salivarius), lactobacillus sake (Lactobacillus sakei) or lactobacillus curvatus (Lactobacillus curvatus). According to a preferred embodiment of the use according to the invention, the lactobacillus is lactobacillus acidophilus.
According to an embodiment of the use according to the invention, the coccus may be selected from enterococcus faecalis (Enterococcus faecalis); streptococcus (Streptococcus), such as Streptococcus thermophilus (Streptococcus thermophilus); lactococcus (lactococcus) such as lactococcus lactis subspecies lactis (Lactococcus Lactis subsp. Lactis), lactococcus lactis subspecies milk fat (Lactococcus Lactis subsp. Cremoris), lactococcus diacetyl subspecies lactis (Lactococcus Lactis subsp. Diacetylactis); leuconostoc (Leuconostoc), such as Leuconostoc mesenteroides (Leuconostoc mesenteroides subsp. Mesenteroides); pediococcus (Pediococcus), such as Pediococcus lactis (Pediococcus acidilactici), pediococcus pentosaceus (Pediococcus pentosaceus); or staphylococci (Staphylococcus) such as one or more of Staphylococcus aureus (Staphylococcus vitulinus), staphylococcus xylosus (Staphylococcus xylosus), and Staphylococcus sarcodactylis (Staphylococcus carnosus). According to a preferred embodiment of the use according to the invention, the coccus is enterococcus faecalis.
According to an embodiment of the use according to the invention, the Bacillus is selected from one or more of Bacillus cereus or Bacillus coagulans (Bacillus coagulans). According to a preferred embodiment of the use according to the invention, the bacillus is bacillus cereus.
Preferably, the bifidobacterium infantis has a collection number of CGMCC No.0460.1; the lactobacillus acidophilus has a strain preservation number of CGMCC No.0460.2; the preservation number of the enterococcus faecalis is CGMCC No.0460.3; the bacillus cereus has a strain preservation number of CGMCC No.0460.4.
According to a specific embodiment of the use according to the invention, the probiotic composition comprises bifidobacterium infantis, lactobacillus acidophilus, enterococcus faecalis and bacillus cereus in a ratio of the number of viable bacteria of (80-120): 8-12): 0.8-1.2.
According to an embodiment of the use of the invention, the disease caused by the allergic reaction may be selected from one or more of systemic allergic diseases, wind mass, allergic rhinitis, allergic asthma or food allergy.
According to an embodiment of the use according to the invention, the allergic reaction may be caused by one or more allergens selected from the group consisting of: pollen allergens, milk allergens, venom allergens, egg allergens, weed allergens, pasture allergens, tree allergens, shrubs allergens, flower allergens, vegetable allergens, cereal allergens, fungal allergens, fruit allergens, berry allergens, nut allergens, seed allergens, soybean allergens, fish allergens, shellfish allergens, seafood allergens, meat allergens, fragrance allergens, insect allergens, mite allergens (e.g., house dust mites), mold allergens, animal allergens, pigeon ticks allergens, helminth allergens, soft coral allergens, animal dander allergens, nematode allergens or rubber tree allergens.
According to an embodiment of the use according to the invention, the allergic reaction may be caused by one or more allergens selected from the group consisting of: inhalant allergens such as pollen particles, dust mite faeces, fungal hyphae and spores, insect venom, animal fur, etc.; food allergens such as milk, eggs, fish, shrimp, crab, shellfish, etc. food proteins or peptides; enzymes such as cysteine proteins in dust mites, and the like.
In a preferred embodiment of the use according to the invention, the allergic reaction is caused by mite allergens.
According to an embodiment of the use according to the invention, the probiotic composition may further comprise Propionibacterium (Propionibacterium), such as one or more of the sub-species Propionibacterium freudenreichii (Propionibacterium freudenreichii subsp. Shermanii), propionibacterium propionicum (Propionibacterium acidipropionici), wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and propionibacteria in a ratio of viable bacteria number of (50-150): 2.5-7.5): 0.5-1.5: 2.5-7.5. According to a preferred embodiment of the use according to the invention, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and propionibacteria in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (4-6).
According to an embodiment of the use of the present invention, the probiotic composition may further comprise one or more of Kluyveromyces (Kluyveromyces), such as Kluyveromyces marxianus (Kluyveromyces marxianus), wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and Kluyveromyces in a ratio of viable bacteria number of (50-150): 5-15): 0.25-0.75. According to a preferred embodiment of the use according to the invention, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces in a ratio of the number of viable bacteria of (80-120): 8-12): 0.8-1.2: (0.4-0.6.
According to an embodiment of the use according to the invention, the medicament may further comprise an allergen preparation.
According to an embodiment of the use of the invention, the ratio of the probiotic composition to the allergen preparation may be (1×10) 5 Up to 1X 10 9 cfu) (20 to 2X 10 5 SQ-U), preferably (1X 10) 6 Up to 1X 10 8 cfu) (1000 to 2X 10 5 SQ-U)。
According to an embodiment of the use according to the invention, the allergen preparation may be selected from one or more of a microbial allergen preparation, a mite allergen preparation, a parasite allergen preparation, a pollen allergen preparation, a xenogeneic animal serum allergen preparation, a pharmaceutical allergen preparation or a chemical agent allergen preparation. According to a preferred embodiment of the use according to the invention, the allergen preparation is a mite allergen preparation, such as a house dust mite allergen preparation or a dust mite allergen preparation.
According to a preferred embodiment of the use according to the invention, the house dust mite allergen preparation is a mite allergen extract, for example an extract of mite bodies, mite body fragments, mite faeces, larvae and/or eggs.
According to a preferred embodiment of the use according to the invention, the probiotic composition is placed separately from the allergen preparation. Further preferably, the probiotic composition is an oral formulation and the allergen formulation is an injection.
In another aspect, the present invention also provides a pharmaceutical composition for preventing and/or treating a disease caused by an allergic reaction, comprising a probiotic composition and an allergen preparation, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of the number of viable bacteria of (50-150): 5-15): 0.5-1.5; and the ratio of the probiotic composition to the allergen preparation is (1×10) 5 Up to 1X 10 9 cfu) (20 to 2X 10 5 SQ-U)。
According to a preferred embodiment of the pharmaceutical composition according to the invention, the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2).
According to a preferred embodiment of the pharmaceutical composition according to the invention, the probiotic composition comprises a total number of viable bacteria not lower than 1 x 10 5 cfu/g. More preferably, the viable count of bifidobacteria in the probiotic composition is at least 1 x 10 6 cfu/g; the viable count of Lactobacillus is at least 1×10 6 cfu/g; the number of living bacteria of the coccus is at least 1×10 6 cfu/g; and the viable count of the bacillus is at least 1 multiplied by 10 5 cfu/g. According to an embodiment of the pharmaceutical composition of the present invention, the bifidobacterium may be selected from one or more of bifidobacterium adolescentis (Bifidobacterium adolescentis), bifidobacterium animalis (Bifidobacterium animalis), bifidobacterium bifidum (Bifidobacterium bifidum), bifidobacterium breve (Bifidobacterium breve), bifidobacterium infantis (Bifidobacterium infantis) or bifidobacterium longum (Bifidobacterium longum). According to a preferred embodiment of the pharmaceutical composition according to the invention, the bifidobacteria are bifidobacteria infantis.
According to an embodiment of the pharmaceutical composition of the invention, the lactobacillus may be selected from one or more of lactobacillus acidophilus (Lactobacillus acidophilus), lactobacillus casei (Lactobacillus casei), lactobacillus crispatus (Lactobacillus crispatus), lactobacillus delbrueckii subspecies bulgaricus (bulgaricus) (Lactobacillus delbrueckii subsp.Bulgarica) (Lactobacillus bulgaricus)), lactobacillus delbrueckii subsp (Lactobacillus delbrueckii subsp.Lactis), lactobacillus fermentum (Lactobacillus fermentium), lactobacillus grignard (Lactobacillus gasseri), lactobacillus helveticus (Lactobacillus helveticus), lactobacillus johnsonii (Lactobacillus johnsonii), lactobacillus paracasei (Lactobacillus paracasei), lactobacillus plantarum (Lactobacillus plantarum), lactobacillus reuteri (Lactobacillus reuteri), lactobacillus rhamnosus (Lactobacillus rhamnosus), lactobacillus salivarius (Lactobacillus salivarius), lactobacillus sake (Lactobacillus sakei) or lactobacillus curvatus (Lactobacillus curvatus). According to a preferred embodiment of the pharmaceutical composition according to the invention, the lactobacillus is lactobacillus acidophilus.
According to an embodiment of the pharmaceutical composition of the invention, the coccus may be selected from enterococcus faecalis (Enterococcus faecalis); streptococcus (Streptococcus), such as Streptococcus thermophilus (Streptococcus thermophilus); lactococcus (lactococcus) such as lactococcus lactis subspecies lactis (Lactococcus Lactis subsp. Lactis), lactococcus lactis subspecies milk fat (Lactococcus Lactis subsp. Cremoris), lactococcus diacetyl subspecies lactis (Lactococcus Lactis subsp. Diacetylactis); leuconostoc (Leuconostoc), such as Leuconostoc mesenteroides (Leuconostoc mesenteroides subsp. Mesenteroides); pediococcus (Pediococcus), such as Pediococcus lactis (Pediococcus acidilactici), pediococcus pentosaceus (Pediococcus pentosaceus); or staphylococci (Staphylococcus) such as one or more of Staphylococcus aureus (Staphylococcus vitulinus), staphylococcus xylosus (Staphylococcus xylosus), and Staphylococcus sarcodactylis (Staphylococcus carnosus). According to a preferred embodiment of the pharmaceutical composition according to the invention, the coccus is enterococcus faecalis.
According to an embodiment of the pharmaceutical composition of the invention, the Bacillus is selected from one or more of Bacillus cereus or Bacillus coagulans (Bacillus coagulans). According to a preferred embodiment of the pharmaceutical composition according to the invention, the bacillus is bacillus cereus.
According to a preferred embodiment of the pharmaceutical composition according to the invention, the probiotic combinationThe ratio of the number of the living bacteria is (80-120), (8-12), and (0.8-1.2) of bifidobacterium infantis, lactobacillus acidophilus, enterococcus faecalis and bacillus cereus; and the ratio of the probiotic composition to the allergen preparation is 1×10 6 Up to 1X 10 8 cfu) (1000 to 2X 10 5 SQ-U)。
Preferably, the bifidobacterium infantis has a collection number of CGMCC No.0460.1; the lactobacillus acidophilus has a strain preservation number of CGMCC No.0460.2; the preservation number of the enterococcus faecalis is CGMCC No.0460.3; the bacillus cereus has a strain preservation number of CGMCC No.0460.4.
According to an embodiment of the pharmaceutical composition of the present invention, the disease caused by the allergic reaction may be selected from one or more of systemic allergic diseases, wind mass, allergic rhinitis, allergic asthma or food allergy.
According to an embodiment of the pharmaceutical composition of the invention, the allergic reaction may be caused by one or more allergens selected from the group consisting of: pollen allergens, milk allergens, venom allergens, egg allergens, weed allergens, pasture allergens, tree allergens, shrubs allergens, flower allergens, vegetable allergens, cereal allergens, fungal allergens, fruit allergens, berry allergens, nut allergens, seed allergens, soybean allergens, fish allergens, shellfish allergens, seafood allergens, meat allergens, fragrance allergens, insect allergens, mite allergens (e.g., house dust mites), mold allergens, animal allergens, pigeon ticks allergens, helminth allergens, soft coral allergens, animal dander allergens, nematode allergens or rubber tree allergens.
According to an embodiment of the pharmaceutical composition according to the invention, the probiotic composition may further comprise Propionibacterium (Propionibacterium), such as one or more of the sub-species Propionibacterium freudenreichii (Propionibacterium freudenreichii subsp. Shermanii), propionibacterium propionicum (Propionibacterium acidipropionici), wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and propionibacteria in a ratio of viable bacteria number of (50-150): 2.5-7.5): 0.5-1.5): 2.5-7.5. In a preferred embodiment of the pharmaceutical composition according to the invention, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and propionibacteria in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (4-6).
According to an embodiment of the pharmaceutical composition of the present invention, the probiotic composition may further comprise Kluyveromyces (Kluyveromyces), such as one or more of Kluyveromyces marxianus (Kluyveromyces marxianus), wherein the probiotic composition comprises a ratio of viable bacteria amounts of (50-150): (50-150): (5-15): (0.25-0.75): (0.25-0.75) bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces. In a preferred embodiment of the pharmaceutical composition according to the invention, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (0.4-0.6).
According to an embodiment of the pharmaceutical composition of the present invention, the allergen preparation may be selected from one or more of a microbial allergen preparation, a mite allergen preparation, a parasite allergen preparation, a pollen allergen preparation, a xenogeneic animal serum allergen preparation, a pharmaceutical allergen preparation or a chemical agent allergen preparation. According to a preferred embodiment of the pharmaceutical composition according to the invention, the allergen preparation is a mite allergen preparation, such as a house dust mite allergen preparation or a dust mite allergen preparation.
In a preferred embodiment of the pharmaceutical composition according to the invention, the probiotic composition is placed separately from the allergen preparation. Further preferably, the probiotic composition is an oral formulation and the allergen formulation is an injection.
In yet another aspect, the present invention also provides a kit for preventing and/or treating a disease caused by an allergic reaction, comprising the pharmaceutical composition of the present invention.
According to a preferred embodiment of the kit according to the invention, the probiotic composition and the allergen preparation are placed separately. Further preferably, the probiotic composition is an oral formulation and the allergen formulation is an injection.
The invention successfully expands the four-linkage or more than four-linkage probiotic composition containing bifidobacterium, lactobacillus, coccus or bacillus from the digestive system field to the new antiallergic treatment field, and has important clinical application significance and value.
The tetrad viable bacteria in the antiallergic probiotic composition has low content and good treatment effect, can remarkably treat anaphylactic reaction, and reduces the probability of anaphylactic diseases of organisms. Meanwhile, the probiotic microecological preparation components contained in the probiotic composition are harmless, and have no side effects on the body. More importantly, the invention also combines the four-combined live bacteria preparation with the allergen preparation, can change the progress of allergic diseases, obviously improves the drug effect of immunotherapy, realizes the optimal treatment means of anaphylactic reaction and can radically achieve the curative effect; in addition, the adoption of the treatment means can greatly shorten the antiallergic treatment period of the tetrad live bacteria preparation or the allergen preparation; meanwhile, the composition can greatly relieve the allergic symptoms under the existing allergen preparation treatment scheme in the process of treating allergic reaction, thereby reducing the occurrence of adverse reactions. In addition, the composition can obviously promote the level of IL-2 while treating anaphylactic reaction, thereby enhancing the immune protection capability of organisms, eliminating the side effect of immunity reduction often accompanied by the existing anaphylactic treatment scheme, and providing a valuable composition for treating clinical antiallergic diseases and enhancing immunity.
Drawings
Embodiments of the present invention are described in detail below with reference to the attached drawing figures, wherein:
FIG. 1 shows the variation of Derp specific IgE in mice after modeling;
FIG. 2 shows changes in Derp specific IgE in mice before/after 4 weeks of challenge;
FIG. 3 shows changes in Derp specific IgE in mice before/after 12 weeks of challenge;
FIG. 4 shows changes in Derp specific IgE in mice before and after challenge for different treatment cycles;
FIG. 5 shows allergy symptom scores for mice treated for 12 weeks;
FIG. 6 shows the changes in secretion of Derp specific cytokines by splenic lymphocytes from mice treated for 12 weeks.
Detailed Description
The following detailed description of the invention is provided in connection with the accompanying drawings that are presented to illustrate the invention and not to limit the scope thereof.
The reagents and preparation methods used in the following examples are described below:
1. house dust mite allergen preparation (AIT): andesda, manufacturer: ALK-Abello A/S. The components are as follows: mite allergen extract (Mites Allergen Product), adjuvants: aluminum hydroxide, sodium chloride, sodium bicarbonate, phenol 5mg/ml, water for injection.
Dosage of administration:
each mouse of house dust mite allergen system is dosed at 10000SQ-U (standard 100000 SQ-U/ml) which is about one tenth of the dose of clinical human.
2. Mite allergen (Derp), manufactured by Zhejiang I WUBio Inc. of Biotech Co., ltd.
3. The preparation method of the single or multiple viable bacteria preparation comprises the following steps:
(1) Dosage of the four-way probiotic suspension for administration of the experimental group:
the strain preservation number of the bifidobacterium infantis is CGMCC No.0460.1; the preservation number of lactobacillus acidophilus is CGMCC No.0460.2; the preservation number of the enterococcus faecalis is CGMCC No.0460.3; the preservation number of the bacillus cereus is CGMCC No.0460.4.
Bifidobacterium infantis (Y) was administered at a concentration of 1.36X 10 per mouse 6 cfu; lactobacillus acidophilus (S) was administered at a concentration of 1.36X 10 per mouse 6 cfu; enterococcus faecalis (F) was administered at a concentration of 1.36X 10 per mouse 5 cfu; bacillus cereus (L) was administered at a concentration of 1.36×10 per mouse 4 cfu (administered dose approximately equal to one tenth of the dose administered to a clinical human).
Example 1 constructionVertical mouse allergy model
SPF-class Balb/c mice, females, 5-8 weeks old were selected for testing. The molding was performed using mite allergens. After the mice were acclimatized, derp (20. Mu.g/ml, 0.1 ml/mouse) was sensitized by subcutaneous injection. The sensitization period is sensitization every other day, and the sensitization is totally 3 times. Mice were sensitized, 1 week apart. Derp (80. Mu.g/ml, 40. Mu.l/mouse) was given to mice by nasal drip. The excitation period was 1 time/day for 7 consecutive days.
After the molding was completed, mice were subjected to facial blood sampling, centrifugation, serum separation, and changes in the Derp-specific IgE in the serum were detected by ELISA, and the results are shown in table 1 and fig. 1.
TABLE 1 variation of Derp specific IgE (MEAN+ -SD) in mice after modeling was completed
| Group of | MEAN | SD | n |
| Normal control group | 0.08 | 0.00 | 10 |
| Model control group | 0.64 | 0.16 | 10 |
The results showed that after mice were sensitized and challenged by de rp, the serum de rp IgE levels of the mice were significantly elevated (P < 0.001) in the model control group compared to the normal control group, indicating successful establishment of the mite allergen-induced mice allergy model.
Example 2 4 week treatment experiment
1. A mouse allergy model was established as in example 1.
2. Model animals were randomly divided into a model control group, an AIT group (andersoprol mite allergen preparation), an SLK group (tetrad probiotic powder suspension), an slk+ait group (combination treatment group: tetrad probiotic powder suspension+andersoprol mite allergen preparation), 10 of each group, alternatively 10 normal mice as normal control groups, and were treated as follows:
a. subcutaneous injection: physiological saline (control group) or house dust mite allergen preparation (experimental group), 0.1 ml/dose (gauge 100000 SQ-U/ml), 1 time per week, continuous treatment for 4 weeks;
b. gastric lavage: physiological saline (control group) or probiotic suspension (experimental group), 0.2 ml/dose, 1 time per day, for 4 weeks.
After 4 weeks of desensitization treatment, the changes in the Derp specific IgE in the serum of each group of mice were detected by ELISA, and the results are shown in Table 2 and FIG. 2.
The results showed that after 4 weeks, the serum de rp IgE remained high in the model control mice compared to the normal control (P < 0.001) (see figure 2).
At the end of 4 weeks of treatment, mice of each group were subjected to Derp nasal drop challenge. The results showed that after challenge, the serum de rp specific IgE was significantly increased (P < 0.001) in each of the remaining mice except for the normal control group, wherein the IgE ratio was changed 3.77-fold after challenge compared to the IgE ratio before challenge in the model control group, the IgE ratio was changed 2.43-fold after challenge compared to the IgE ratio before challenge in the AIT group, the IgE ratio was changed 3.28-fold after challenge in the SLK group, and the IgE ratio was changed 2.22-fold after challenge in the slk+ait group. Mice in the AIT group (P < 0.01), SLK group (P < 0.05), slk+ait group (P < 0.01) had significantly reduced serum Derp IgE compared to the model control group (see figure 2). As can be seen from table 2, the allergic treatment was followed by 4 weeks of challenge, and it can be seen from the IgE variation level of each experimental group that all of the AIT group, the SLK group, and the slk+ait group had a certain desensitizing effect, wherein the slk+ait group had a better effect than the AIT group and the SLK group.
TABLE 2 changes in Derp specific IgE (MEAN+ -SD) in mice after 4 weeks of treatment
And (3) injection: (1) P <0.05, < P <0.01, < P <0.001, compared to the model control group; (2) After challenge, compared to before challenge, #p <0.05, #p <0.01, #p <0.001.
Example 3 12 week treatment experiment
1. A mouse allergy model was established as in example 1.
2. Model animals were randomly divided into model control group, AIT group, SLK, slk+ait group, 10 of each group, alternatively 10 normal mice as normal control group, and the following treatments were performed:
a. subcutaneous injection: physiological saline (control group) or house dust mite allergen preparation (experimental group), 0.1 ml/dose (gauge 100000 SQ-U/ml), 1 time per week, continuous treatment for 12 weeks;
b. gastric lavage: physiological saline (control group) or probiotic suspension (experimental group), 0.2 ml/dose, 1 time per day for 12 weeks.
3. Therapeutic results
(1) Changes in Derp-specific IgE in mice after 12 weeks of treatment
After 12 weeks of desensitization treatment, the changes in the Derp specific IgE in the serum of each group of mice were detected by ELISA, and the results are shown in Table 3 and FIG. 3.
At the end of 12 weeks of treatment, mice of each group were subjected to Derp nasal drop challenge. The results showed that after challenge, the serum de rp specific IgE was significantly elevated (P < 0.001) for each of the remaining groups of mice except the normal control group and the slk+ait group (fig. 3). Wherein the IgE ratio of the model control group after excitation is 3.71 times changed, the IgE ratio of the AIT group after excitation is 2.04 times changed, the IgE ratio of the SLK group after excitation is 1.83 times changed, and the IgE ratio of the SLK+AIT group after excitation is 1.57 times changed. Mice from the SLK and SLK+AIT groups showed significantly reduced serum Derp-specific IgE compared to the model control group (SLK group: P <0.01; SLK+AIT group: P < 0.001) (FIG. 3). Combining the effects of each group of treatments after 12 weeks of stimulation, it can be seen (table 3) that all of the AIT, SLK, slk+ait groups had varying degrees of desensitization, with the slk+ait groups being seen from the level of specific IgE, already achieved desensitization; the desensitization effect of SLK group is slightly better than AIT group.
TABLE 3 changes in Derp specific IgE (MEAN+ -SD) in mice after 12 weeks of treatment
Note that: (1) P <0.05, < P <0.01, < P <0.001, compared to the model control group; (2) After challenge, compared to before challenge, #p <0.05, #p <0.01, #p <0.001.
Figure 4 shows the change in de rp specific IgE in mice before and after challenge for 4 weeks, 12 weeks of desensitization treatment with different treatment cycles. As can be seen from the upper graph (a: pre-challenge) of fig. 4, the pre-challenge AIT group, SLK group de rp-specific IgE is on an ascending trend with increasing treatment cycle, indicating that the therapeutic effect was not achieved; the slk+ait group remained stable IgE during the course of treatment, indicating that therapeutic effect was achieved. As can be seen from the lower panel of fig. 4 (B: post challenge), igE in AIT group mice after challenge still showed an increasing trend during treatment; whereas IgE in mice of SLK group, SLK+AIT group, was decreased during the course of treatment. At 12 weeks of treatment, slk+ait group mice had significantly reduced serum de rp specific IgE (P < 0.01) compared to the model control group. It was shown that SLK in combination with AIT helps to alleviate IgE elevation.
(2) Grading of allergic symptoms in mice after 12 weeks of treatment
After 12 weeks of treatment, mice in each group were subjected to continuous 7-day nasal drop challenge, and mice allergic symptoms were counted on day 7. Figure 5 shows allergy symptoms scores for mice treated for 12 weeks. As can be seen from fig. 5, the slk+ait group had significantly reduced allergy symptoms score (P < 0.01) compared to the model control group, and the mice were significantly less scratched and wheezing; the allergic reaction symptoms of the allergen preparation AIT group are obviously more serious than those of the four-combined live bacteria preparation SLK group and SLK+AIT group, so that the four-combined live bacteria preparation SLK group and SLK+AIT group can obviously reduce the occurrence of the allergic reaction symptoms, thereby reducing the incidence rate of adverse reactions.
(3) Mouse spleen lymphocytes secrete Derp specific IL-2 after 12 weeks of treatment
After 12 weeks of treatment, mice were subjected to eyeball-picking, blood sampling, neck-removing and sacrifice to obtain spleen lymphocytes. The secretion of Derp specific IL-2 by mouse spleen lymphocytes was detected by enzyme-linked immunosorbent assay (enzyme-linked immunospot, ELISPOT) and the results are shown in FIG. 6. As can be seen from fig. 6 (a), the levels of Derp-specific IL-2 secretion by mouse spleen lymphocytes can be significantly increased after the combination therapy of SLK and AIT. The results indicate that the SLK group and SLK+AIT group can achieve desensitization treatment by regulating the balance of Th1/Th 2.
The test results show that the probiotic composition or the probiotic composition combined mite allergen preparation can effectively treat or relieve anaphylactic reaction in the treatment effect, and meanwhile, the probiotic composition combined mite allergen preparation can achieve the treatment effect faster than the mite allergen preparation alone, and is more convenient to treat. In addition, on the allergic symptom score, the probiotic combination or the probiotic composition combined with the mite allergen preparation can obviously reduce the allergic symptom occurrence and thus reduce the adverse reaction incidence. The probiotic composition or the combination of the probiotic composition and the mite allergen preparation can achieve the treatment effect better and faster than the single treatment of the mite allergen preparation, and is expected to reduce the treatment expense of patients.
Claims (15)
1. Use of a probiotic composition comprising bifidobacteria, lactobacilli, cocci and bacilli for the manufacture of a medicament for the prevention and/or treatment of diseases caused by allergic reactions.
2. Use according to claim 1, wherein the probiotic composition comprises live bacteriaThe total number is not less than 1×10 5 cfu/g; preferably, the viable count of bifidobacteria in the probiotic composition is at least 1 x 10 6 cfu/g; the viable count of Lactobacillus is at least 1×10 6 cfu/g; the number of living bacteria of the coccus is at least 1×10 6 cfu/g; and the viable count of the bacillus is at least 1 multiplied by 10 5 cfu/g。
3. The use according to claim 1, wherein the bifidobacterium is selected from one or more of bifidobacterium adolescentis, bifidobacterium animalis, bifidobacterium bifidum, bifidobacterium breve, bifidobacterium infantis or bifidobacterium longum; preferably, the bifidobacterium is bifidobacterium infantis;
the lactobacillus is selected from one or more of lactobacillus acidophilus, lactobacillus casei, lactobacillus crispatus, lactobacillus bulgaricus, lactobacillus delbrueckii subspecies lactis, lactobacillus fermentum, lactobacillus grignard, lactobacillus helveticus, lactobacillus johnsonii, lactobacillus paracasei, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus salivarius, lactobacillus sake or lactobacillus curvatus; preferably, the lactobacillus is lactobacillus acidophilus;
The coccus is selected from enterococcus faecalis; streptococci, such as streptococcus thermophilus; lactococcus, such as lactococcus lactis subspecies lactis, lactococcus lactis subspecies cremoris, lactococcus lactis diacetyl subspecies lactis; leuconostoc, such as Leuconostoc mesenteroides subspecies; pediococcus such as Pediococcus acidilactici, pediococcus pentosaceus; staphylococci such as one or more of staphylococcus calf, staphylococcus xylosus, staphylococcus sarcodactylis; preferably, the coccus is enterococcus faecalis;
the bacillus is selected from one or more of bacillus cereus and bacillus coagulans; preferably, the bacillus is bacillus cereus;
preferably, the bifidobacterium infantis has a collection number of CGMCC No.0460.1; the lactobacillus acidophilus has a strain preservation number of CGMCC No.0460.2; the preservation number of the enterococcus faecalis is CGMCC No.0460.3; the bacillus cereus has a strain preservation number of CGMCC No.0460.4.
4. The use according to any one of claims 1 to 3, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of viable bacteria number of (50-150): (50-150): (5-15): (0.5-1.5);
preferably, the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2);
More preferably, the probiotic composition comprises bifidobacterium infantis, lactobacillus acidophilus, enterococcus faecalis and bacillus cereus in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2).
5. The use according to any one of claims 1 to 4, wherein the probiotic composition further comprises propionibacteria, such as one or more of the sub-species propionibacterium freudenreichii, propionibacterium propionicum, wherein the probiotic composition comprises bifidobacterium, lactobacillus, cocci, bacillus and propionibacteria in a ratio of viable bacteria count of (50-150): 2.5-7.5: 0.5-1.5: 2.5-7.5; preferably, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and propionibacteria in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (4-6); and/or
The probiotic composition further comprises kluyveromyces, such as one or more of kluyveromyces marxianus, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces in a ratio of viable count of (50-150): (50-150): (5-15): (0.25-0.75): (0.25-0.75); preferably, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (0.4-0.6).
6. The use according to any one of claims 1 to 5, wherein the medicament further comprises an allergen preparation selected from one or more of a microbial allergen preparation, a mite allergen preparation, a parasite allergen preparation, a pollen allergen preparation, a xenogeneic animal serum allergen preparation, a pharmaceutical allergen preparation or a chemical agent allergen preparation; most preferably, the allergen preparation is a mite allergen preparation, such as a house dust mite allergen preparation or a dust mite allergen preparation;
preferably, the ratio of the probiotic composition to allergen preparation is (1×10) 5 Up to 1X 10 9 cfu) (20 to 2X 10 5 SQ-U), preferably (1X 10) 6 Up to 1X 10 8 cfu) (1000 to 2X 10 5 SQ-U)。
7. The use according to any one of claims 1 to 6, wherein the disease caused by an allergic reaction is selected from one or more of systemic allergic disease, wind mass, allergic rhinitis, allergic asthma or food allergy;
preferably, the allergic reaction is caused by one or more allergens selected from the group consisting of: pollen allergens, milk allergens, venom allergens, egg allergens, weed allergens, pasture allergens, tree allergens, shrub allergens, flower allergens, vegetable allergens, cereal allergens, fungal allergens, fruit allergens, berry allergens, nut allergens, seed allergens, soybean allergens, fish allergens, shellfish allergens, seafood allergens, meat allergens, spice allergens, insect allergens, mite allergens, mould allergens, animal allergens, pigeon ticks allergens, helminth allergens, soft coral allergens, animal dander allergens, nematode allergens or rubber tree allergens; preferably, the allergic reaction is caused by a mite allergen.
8. A pharmaceutical composition for preventing and/or treating a disease caused by an allergic reaction, comprising a probiotic composition and an allergen preparation, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of viable bacteria count of (50-150): 5-15): 0.5-1.5.
9. The pharmaceutical composition of claim 8, wherein the allergen formulation is selected from one or more of a microbial allergen formulation, a mite allergen formulation, a parasite allergen formulation, a pollen allergen formulation, a xenogeneic animal serum allergen formulation, a pharmaceutical allergen formulation, or a chemical agent allergen formulation; most preferably, the allergen preparation is a mite allergen preparation, such as a house dust mite allergen preparation or a dust mite allergen preparation;
preferably, the ratio of the probiotic composition to the allergen preparation is (1×10) 5 Up to 1X 10 9 cfu) (20 to 2X 10 5 SQ-U), more preferably the ratio of the probiotic composition to the allergen preparation is (1X 10) 6 Up to 1X 10 8 cfu) (1000 to 2X 10 5 SQ-U)。
10. The pharmaceutical composition of claim 8, wherein the probiotic composition comprises a total number of viable bacteria of not less than 1 x 10 5 cfu/g; preferably, the viable count of bifidobacteria in the probiotic composition is at least 1 x 10 6 cfu/g; the viable count of Lactobacillus is at least 1×10 6 cfu/g; the number of living bacteria of the coccus is at least 1×10 6 cfu/g; and the viable count of the bacillus is at least 1 multiplied by 10 5 cfu/g。
11. The pharmaceutical composition of claim 8, wherein the bifidobacterium is selected from one or more of bifidobacterium adolescentis, bifidobacterium animalis, bifidobacterium bifidum, bifidobacterium breve, bifidobacterium infantis, or bifidobacterium longum; preferably, the bifidobacterium is bifidobacterium infantis;
the lactobacillus is selected from one or more of lactobacillus acidophilus, lactobacillus casei, lactobacillus crispatus, lactobacillus bulgaricus, lactobacillus delbrueckii subspecies lactis, lactobacillus fermentum, lactobacillus grignard, lactobacillus helveticus, lactobacillus johnsonii, lactobacillus paracasei, lactobacillus plantarum, lactobacillus reuteri, lactobacillus rhamnosus, lactobacillus salivarius, lactobacillus sake or lactobacillus curvatus; preferably, the lactobacillus is lactobacillus acidophilus;
the coccus is selected from enterococcus faecalis; streptococci, such as streptococcus thermophilus; lactococcus, such as lactococcus lactis subspecies lactis, lactococcus lactis subspecies cremoris, lactococcus lactis diacetyl subspecies lactis; leuconostoc, such as Leuconostoc mesenteroides subspecies; pediococcus such as Pediococcus acidilactici, pediococcus pentosaceus; staphylococci such as one or more of staphylococcus calf, staphylococcus xylosus, staphylococcus sarcodactylis; preferably, the coccus is enterococcus faecalis;
The bacillus is selected from one or more of bacillus cereus and bacillus coagulans; preferably, the bacillus is bacillus cereus;
preferably, the bifidobacterium infantis has a collection number of CGMCC No.0460.1; the lactobacillus acidophilus has a strain preservation number of CGMCC No.0460.2; the preservation number of the enterococcus faecalis is CGMCC No.0460.3; the bacillus cereus has a strain preservation number of CGMCC No.0460.4.
12. The pharmaceutical composition according to any one of claims 8 to 11, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci and bacilli in a ratio of viable bacteria number of (80-120): (80-120): (8-12): (0.8-1.2); preferably, the probiotic composition comprises bifidobacterium infantis, lactobacillus acidophilus, enterococcus faecalis and bacillus cereus in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2).
13. The pharmaceutical composition according to any one of claims 8 to 12, wherein the probiotic composition further comprises propionibacteria, such as one or more of the sub-species propionibacterium freudenreichii, propionibacterium propionicum, wherein the probiotic composition comprises bifidobacterium, lactobacillus, cocci, bacillus and propionibacteria in a ratio of viable bacteria count of (50-150): 2.5-7.5: 0.5-1.5: 2.5-7.5; preferably, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and propionibacteria in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (4-6); and/or
The probiotic composition further comprises kluyveromyces, such as one or more of kluyveromyces marxianus, wherein the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces in a ratio of viable count of (50-150): (50-150): (5-15): (0.25-0.75): (0.25-0.75); preferably, the probiotic composition comprises bifidobacteria, lactobacilli, cocci, bacilli and kluyveromyces in a ratio of the number of viable bacteria of (80-120): (80-120): (8-12): (0.8-1.2): (0.4-0.6).
14. A kit for preventing and/or treating a disease caused by an allergic reaction, comprising the pharmaceutical composition according to any one of claims 8 to 13.
15. Kit according to claim 14, wherein the probiotic composition and the allergen preparation are placed separately, preferably the probiotic composition is an oral preparation and the allergen preparation is an injection.
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