Use of lactic acid bacterial strains for the treatment of atopic dermatitis

Abstract

The present application relates to the use of lactic acid bacterial strains for treating atopic dermatitis. The present application discloses that a lactic acid bacterial strain can be used for treating atopic dermatitis, wherein the lactic acid bacterial strain is selected from the group consisting of: Salivaria lactis AP-32 (CCTCC M 2011127), Bifidobacterium animalis lactis subsp. CP-9 (CCTCC M 2014588), and combinations thereof.

Description

Technical Field

[0001] This invention relates to the use of lactic acid bacteria strains to treat atopic dermatitis, wherein the lactic acid bacteria strains are selected from the group consisting of: Lactobacillus salivarius subsp. salicinius AP-32 (CCTCC M 2011127), Bifidobacterium animalis subsp. lactis CP-9 (CCTCC M 2014588), and combinations thereof. Background Technology

[0002] Atopic dermatitis (AD) [also known as atopic eczema] is a common and recurrent allergic skin disease, believed to be mainly caused by a Th1 / Th2 dysbalance. A significant decrease in the expression of Th1 cytokines [including interferon-γ (IFN-γ)] and a significant increase in the expression of Th2 cytokines [including interleukin-13 (IL-13)] can be observed.

[0003] Common symptoms include itchy skin, redness and swelling, desquamation, cracking, and crust formation. In infants and young children, the face is the most common site, while in children and adults, the flexor surfaces of the knees and elbows are more common. In addition, many patients also develop allergic rhinitis, asthma, and allergic conjunctivitis.

[0004] Currently, the main medications used clinically to treat atopic dermatitis include oral antihistamines and antibiotics, as well as topical corticosteroids and immunosuppressants. However, these medications can cause side effects and drug resistance, leading to persistent symptoms and relapses even after prolonged treatment. Therefore, researchers in this field are dedicated to developing drugs that can effectively treat atopic dermatitis without causing unwanted side effects.

[0005] Lactic acid bacteria (LAB) are generally recognized as safe (GRAS) and are well-known and widely used probiotics. Common lactic acid bacteria include: Lactobacillus, Lactococcus, Pediococcus, Enterococcus, Streptococcus, Bifidobacterium, Bacillus, and Leuconostoc.

[0006] Current research focuses on the application of lactic acid bacteria strains in improving atopic dermatitis. For example, in Isolauri E et al. (2000), Clin. Exp. Allergy, 30:1604-1610, Isolauri E et al. added Lactobacillus strain GG (ATCC 53103) [i.e. Lactobacillus rhamnosus GG (LGG)] and Bifidobacterium lactis BB-12 [i.e. Bifidobacterium animalis subsp. lactis BB-12] to an extensively hydrolyzed whey formula and fed it to infants exhibiting atopic dermatitis during breastfeeding. The results showed that both Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB-12 effectively improved atopic dermatitis.

[0007] Although the aforementioned literature has been reported, there is still a need in this field to screen for lactic acid bacteria that can effectively treat atopic dermatitis for use by industry. Summary of the Invention

[0008] In this invention, the applicant discovered through experiments that Lactobacillus salivarius subsp. salicinius AP-32 (CCTCC M 2011127) and Bifidobacterium animalis subsp. lactis CP-9 (CCTCC M 2014588) can both effectively improve the Th1 / Th2 dysbalance in patients with atopic dermatitis (AD), and produce a synergistic effect when used in combination at a bacterial count ratio of 1:0.11 to 1:9.

[0009] Therefore, in a first aspect, the present invention provides the use of a lactic acid bacteria strain for preparing a composition for treating atopic dermatitis, wherein the lactic acid bacteria strain is selected from the group consisting of: Lactobacillus salivarius AP-32, Bifidobacterium animalis subsp. lactis CP-9, and combinations thereof.

[0010] Preferably, the lactic acid bacteria strain is a combination containing Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9.

[0011] Preferably, the ratio of Lactobacillus salivarius AP-32 to Bifidobacterium animalis subsp. lactis CP-9 falls within the range of 1:0.11 to 1:9.

[0012] More preferably, the bacterial count ratio is 1:9.

[0013] Preferably, the composition is a pharmaceutical composition.

[0014] More preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[0015] More preferably, the pharmaceutical composition is in a dosage form for oral or topical administration.

[0016] In a second aspect, the present invention provides a method for treating atopic dermatitis, comprising administering to an individual in need the lactic acid bacteria strain described above. Attached Figure Description

[0017] Figure 1 The image shows the IFN-γ concentration (pg / mL) of each group of PBMCs after treatment with different lactic acid bacteria strains. "This indicates that when compared with single-strain control group 1, p < 0.001; and " "Indicates that when compared with single-strain control group 2, p < 0.01;

[0018] Figure 2 The image shows the IL-10 concentration (pg / mL) of each group of PBMCs after treatment with different lactic acid bacteria strains. "This indicates that when compared with single-strain control group 1, p < 0.001; and " "Indicates that when compared with single-strain control group 2, p < 0.001;

[0019] Figure 3 The image shows the IL-13 concentration (pg / mL) of each group of PBMCs after treatment with different lactic acid bacteria strains. "This indicates that when compared with single-strain control group 1, p < 0.001; and " "Indicates that when compared with single-strain control group 2, p < 0.001;

[0020] Figure 4 The image shows the IFN-γ concentration (pg / mL) of each group of PBMCs after treatment with different lactic acid bacteria strains. "This indicates that when compared with single-strain group 1, p < 0.001;" "This indicates that when compared with group 2 (single bacteria), p < 0.001; and " "and" "" indicates that when compared with the compound bacterial group 4, p < 0.01 and p < 0.001, respectively;

[0021] Figure 5 The image shows the IL-10 concentration (pg / mL) of each group of PBMCs after treatment with different lactic acid bacteria strains. "This indicates that when compared with single-strain group 1, p < 0.001; and " "This indicates that when compared with group 2 (single bacteria), p < 0.001; and"

[0022] Figure 6 The image shows the IL-13 concentration (pg / mL) of each group of PBMCs after treatment with different lactic acid bacteria strains. "This indicates that when compared with single-strain group 1, p < 0.05; and " "" indicates that when compared with single bacterial group 2, p < 0.05. Detailed Implementation

[0023] For the purposes of this instruction manual, it will be clearly understood that the word "comprising" means "including but not limited to", and the word "comprises" has the corresponding meaning.

[0024] Unless otherwise defined, all technical and scientific terms used herein have the meaning commonly understood by those skilled in the art. Those skilled in the art will recognize many similar or equivalent methods and materials that can be used to practice the invention. Of course, the invention is by no means limited to the methods and materials described.

[0025] This invention provides the use of a lactic acid bacteria strain for preparing a composition for treating atopic dermatitis, wherein the lactic acid bacteria strain is selected from the group consisting of: Lactobacillus salivarius subsp. salicinius AP-32 (CCTCC M 2011127), Bifidobacterium animalis subsp. lactis CP-9 (CCTCC M 2014588), and combinations thereof.

[0026] As used in this article, “treating” or “treatment” means preventing, reducing, alleviating, ameliorating, relieving, or controlling one or more clinical signs of a disease or disorder, as well as lowering, stopping, or reversing the progression of the severity of a condition or symptom that is being treated.

[0027] In a preferred embodiment of the present invention, the lactic acid bacteria strain is a combination containing Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9.

[0028] Preferably, the ratio of Lactobacillus salivarius AP-32 to Bifidobacterium animalis subsp. lactis CP-9 falls within the range of 1:0.11 to 1:9. More preferably, this ratio is 1:9.

[0029] According to the present invention, the combination of lactic acid bacteria strains may have a range of 10 7 Up to 10 12 The bacterial concentration within CFU / mL is preferably 10. 8 Up to 10 10 CFU / mL. In a preferred embodiment of the invention, the bacterial concentration of the lactic acid bacteria strain combination is 10. 9 CFU / mL.

[0030] According to the present invention, Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 can be live or dead bacteria, concentrated or non-concentrated, liquid, paste, semi-solid, or solid [e.g., pellets, granules, or powder], and can be heat-inactivated, frozen, dried, or freeze-dried [e.g., in freeze-dried or spray / fluid bed dried form].

[0031] According to the present invention, the composition may be a pharmaceutical composition.

[0032] According to the present invention, the pharmaceutical composition may be in a dosage form suitable for oral administration or topical administration.

[0033] According to the present invention, the pharmaceutical composition may further comprise a pharmaceutically acceptable carrier widely used in pharmaceutical manufacturing techniques. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the following: solvent, buffer, emulsifier, suspending agent, decomposer, disintegrating agent, dispersing agent, binding agent, excipient, stabilizing agent, chelating agent, diluent, gelling agent, preservative, wetting agent, lubricant, absorption delaying agent, liposome, and the like. The selection and quantity of these agents fall within the scope of expertise and practice of those skilled in the art.

[0034] According to the present invention, the pharmaceutical composition can be manufactured into a dosage form suitable for oral administration using techniques known to those skilled in the art, including, but not limited to: sterile powders, tablets, troche, lozenges, pellets, capsules, dispersible powders or granules, solutions, suspensions, emulsions, syrups, elixirs, slurries, and the like.

[0035] According to the present invention, the pharmaceutical composition can also be manufactured into an external preparation suitable for topical application to the skin using techniques known to those skilled in the art, including, but not limited to: emulsion, gel, ointment, cream, patch, liniment, powder, aerosol, spray, lotion, serum, paste, foam, drop, suspension, salve, and bandage.

[0036] According to the present invention, the external formulation is prepared by mixing the pharmaceutical composition of the present invention with a base known to those skilled in the art.

[0037] According to the present invention, the substrate may contain one or more additives selected from the following: water, alcohols, glycols, hydrocarbons [such as petroleum jelly and white petrolatum], waxes [such as paraffin and yellow wax], preserving agents, antioxidants, surfactants, absorption enhancers, stabilizing agents, and gelling agents [such as carbomer]. ® 941 (carbopol) ®941), microcrystalline cellulose and carboxymethyl cellulose, active agents, humectants, odor bsorbers, fragrances, pH adjusting agents, chelating agents, emulsifiers, occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, antiirritants, colorants, and propellants, etc. The selection and quantity of these additives fall within the scope of expertise and routine practice of those skilled in the art.

[0038] According to the present invention, the pharmaceutical composition may be used in combination with one or more drugs selected from the group consisting of: anti-inflammatory agents, immunosuppressive agents, antihistamines, antiviral agents, wound-healing agents, antipruritic agents, anti-dry skin agents, humectants, and skin nutrients.

[0039] The present invention also provides a method for treating atopic dermatitis, comprising administering, to an individual in need, the lactic acid bacteria strain described above.

[0040] As used herein, the terms “administration” and “administration” may be used interchangeably and mean introducing, providing, or delivering a predetermined active ingredient to an individual by any suitable means to perform its intended effect.

[0041] As used in this article, the term "subject" refers to any mammal of interest, such as humans, monkeys, cows, sheep, horses, pigs, goats, dogs, cats, mice, and rats.

[0042] According to the present invention, the dosage and frequency of administration of the lactic acid bacteria strain will vary depending on the following factors: the severity of the condition to be improved, the route of administration, and the age, physical condition, and response of the individual to be improved. Generally, the lactic acid bacteria strain can be administered orally or locally in a single dose or in multiple doses.

[0043] The present invention will be further described with reference to the following embodiments, but it should be understood that these embodiments are for illustrative purposes only and should not be construed as limiting the implementation of the present invention.

[0044] <Example>

[0045] General experimental materials:

[0046] 1. Lactic acid bacteria strains:

[0047] The lactic acid bacteria strains used for efficacy evaluation in the following examples have been deposited at the Bioresource Collection and Research Center (BCRC) of the Food Industry Research and Development Institute (FIRDI) in Taiwan (No. 331, Food Rd., Hsinchu City, Taiwan, China 300), and are publicly available. In addition, these lactic acid bacteria strains have also been deposited at the China Center for Type Culture Collection (CCTCC) in accordance with the provisions of the Budapest Treaty. For clarity, relevant information for each lactic acid bacteria strain (including: scientific name, relevant source, accession number, and accession date) has been compiled in Table 1 below.

[0048] Table 1. Relevant information for each lactic acid bacteria strain

[0049]

[0050] For comparison, the following commercially available lactic acid bacteria strains with efficacy in improving atopic dermatitis (AD) were also used: Lactobacillus rhamnosus GG (LGG) (corresponding to ATCC 53103) and Bifidobacterium animalis subsp. lactis BB-12 (corresponding to DSM 15954), purchased from Chr. Hansen A / S, Denmark.

[0051] 2. Preparation of cultures of lactic acid bacteria strains:

[0052] First, the four lactic acid bacteria strains described in item 1 above were inoculated into MRS broth medium (Difco, Cat. No. 288130) supplemented with 0.05% cysteine ​​and cultured in an incubator (37°C, 5% CO2) for 24 hours to activate the strains. Next, the activated strains were inoculated into MRS broth medium at a 2% (v / v) inoculation rate and cultured in an incubator (37°C, 5% CO2) for 24 hours. Afterwards, the resulting cultures were adjusted to a concentration of 1×10⁻⁶ using MRS broth medium. 9 The bacterial concentration of CFU / mL (using plate counting medium for bacterial count) was used to obtain cultures of each lactic acid bacteria strain.

[0053] 3. Preparation of peripheral blood mononuclear cells (PBMCs):

[0054] First, peripheral blood was collected from 10 patients (8 males and 2 females, aged 13 years or younger) clinically diagnosed with atopic dermatitis, recruited by Kaohsiung Chang Gung Memorial Hospital. This was conducted according to procedures approved by the Institutional Review Board (IRB) of Kaohsiung Chang Gung Memorial Hospital and in accordance with the current revised version of the Declaration of Helsinki. Informed consent was obtained from all participants after a full explanation of the experiment.

[0055] Next, the obtained peripheral blood was added to Ficoll-Paque™ PLUS centrifuge tubes and subjected to density-gradient centrifugation at 720g for 30 minutes at 4°C, after which the lymphocyte layer was harvested. The red blood cells were then lysed using RBC lysis buffer. The resulting PBMCs were then adjusted to a concentration of 4 × 10⁻⁶ cells / mL using RPMI 1640 medium (Gibco) containing 10% fetal bovine serum (FBS). 6 Cells / mL for later use.

[0056] General experimental methods:

[0057] 1. Statistical analysis:

[0058] In the examples below, each group's experiment was repeated three times, and the experimental data are expressed as mean ± standard error of the mean (SEM). All data were analyzed using Student's t-test to assess differences between groups. A statistically significant result was defined as p < 0.05.

[0059] Example 1. Evaluation of the efficacy of lactic acid bacteria strains in the treatment of atopic dermatitis

[0060] A. Treating PBMCs cells with lactic acid bacteria strains:

[0061] First, the PBMCs obtained according to item 3 of the "General Experimental Materials" section above were divided into 5 groups, including 1 pathological control group, 2 single-bacterial experimental groups (i.e., single-bacterial experimental groups 1 and 2), and 2 single-bacterial comparison groups (i.e., single-bacterial comparison groups 1 and 2). Each group of cells was 4 × 10⁻⁶ cells / year. 5 Cells / wells were cultured in 96-well plates containing 0.12 mL of RPMI 1640 medium [with 10% FBS and 1% penicillin-streptomycin (PS)] and incubated in an incubator (37°C, 5% CO2) for 24 hours.

[0062] Next, each group was replaced with fresh culture medium. Then, according to Table 2 below, appropriate amounts of cultures of the four lactic acid bacteria strains obtained in item 2 of "General Experimental Materials" were added to the corresponding groups, so that each single-strain experimental group and single-strain control group contained 4 × 10⁻⁶ bacteria. 6 The number of lactic acid bacteria in CFU. The cell cultures of the pathological control group were not treated in any way.

[0063] Table 2. Cultures of lactic acid bacteria strains treated in each group

[0064]

[0065] After culturing the cells in each group in an incubator (37℃, 5% CO2) for 48 hours, the resulting culture was centrifuged at 3,000 rpm for 10 minutes, and the supernatant was collected and used for the analysis in item B below.

[0066] B. Concentration determination of interferon-γ (IFN-γ), interleukin-10 (IL-10), and interleukin-13 (IL-13):

[0067] The concentrations (pg / mL) of IFN-γ, IL-10, and IL-13 in the supernatant of each group were determined using the IFN-γ ELISA kit (ThermoFisher, catalog number KHC4021), the IL-10 ELISA kit (ThermoFisher, catalog number BMS215-2), and the IL-13 ELISA kit (ThermoFisher, catalog number BMS231-3) according to the manufacturer's instructions.

[0068] Then, analyze the obtained experimental data according to the method described in item 1 of the "General Experimental Methods" section above.

[0069] Figure 1 and Figure 2 The concentrations of IFN-γ and IL-10 in each group of PBMCs after treatment with different strains of lactic acid bacteria are shown separately. Figure 1 and Figure 2It is evident that, compared to the pathological control group, the concentrations of IFN-γ and IL-10 in each single-bacterial experimental group and the single-bacterial comparison group were significantly increased. In particular, the concentrations of IFN-γ and IL-10 in each single-bacterial experimental group were significantly higher than those in each single-bacterial comparison group. This indicates that Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 can effectively induce Th1 cells to secrete IFN-γ and IL-10 in human PBMCs, and this induction effect is superior to that of existing lactic acid bacteria strains used to improve atopic dermatitis.

[0070] Figure 3 This shows the IL-13 concentrations of PBMCs in each group after treatment with different strains of lactic acid bacteria. Figure 1 It is evident that, compared to the pathological control group, the IL-13 concentrations in each single-bacterial experimental group and the single-bacterial comparison group were significantly reduced. In particular, the IL-13 concentrations in each single-bacterial experimental group were significantly lower than those in each single-bacterial comparison group. This indicates that Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 can effectively inhibit the secretion of IL-13 by Th2 cells in human PBMCs, and this inhibitory effect is superior to that of lactic acid bacteria strains known to be used to improve atopic dermatitis.

[0071] These experimental results show that both Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 can achieve therapeutic effects on atopic dermatitis by improving the Th1 / Th2 dysbalance.

[0072] Example 2. Evaluation of the efficacy of combinations of lactic acid bacteria strains in the treatment of atopic dermatitis

[0073] A. Treating PBMCs cells with lactic acid bacteria strains:

[0074] First, the PBMCs obtained according to item 3 of the "General Experimental Materials" section above were divided into 8 groups, including 1 pathological control group, 2 single-bacterial groups (i.e., single-bacterial groups 1 and 2), and 5 complex-bacterial groups (i.e., complex-bacterial groups 1 to 5). Each group of cells was 4 × 10⁻⁶ cells long. 5 Cells / wells were cultured in 96-well plates containing 0.12 mL of RPMI 1640 medium (with 10% FBS and 1% penicillin-streptomycin) and incubated in an incubator (37°C, 5% CO2) for 24 hours.

[0075] Next, each group was replaced with fresh culture medium. Then, appropriate amounts of cultures of *Lactobacillus salivarius* AP-32 and *Bifidobacterium animalis* subsp. *lactobacter* CP-9, obtained in item 2 of the "General Experimental Materials" section above, were added to single-strain groups 1 and 2, respectively, so that each single-strain group contained 4 × 10⁻⁶ bacteria. 6 The number of lactic acid bacteria in CFU, and according to Table 3 below, cultures of *Lactobacillus salivarius* AP-32 and *Bifidobacterium animalis* subsp. *lactobacter* CP-9 were mixed at different ratios and added to compound bacterial groups 1 to 5 respectively, so that each compound bacterial group contained 4 × 10⁻⁶ bacteria. 6 The number of lactic acid bacteria in CFU. The cell cultures of the pathological control group were not treated in any way.

[0076] Table 3. Ratio of Lactic Acid Bacteria Strains Treated in Each Compound Microbial Group

[0077]

[0078] After culturing the cells in each group in an incubator (37°C, 5% CO2) for 48 hours, the resulting culture was centrifuged at 3,000 rpm for 10 minutes, and the supernatant was collected and used for the analysis in item B below.

[0079] B. Concentration determination of IFN-γ:

[0080] The IFN-γ ELISA kit (ThermoFisher, catalog number KHC4021) was used to determine the IFN-γ concentration (pg / mL) in the supernatant of each group according to the manufacturer's instructions. The experimental data were then analyzed according to the method described in section 1 of the "General Experimental Methods" above. The results are shown below. Figure 1 middle.

[0081] Figure 4 This shows the IFN-γ concentrations of PBMCs in each group after treatment with different lactic acid bacteria strains. Figure 4 It is evident that the IFN-γ concentrations in each single-bacterial group were slightly higher than those in the pathological control group, while the IFN-γ concentrations in compound bacterial groups 1 and 5 were slightly lower than those in single-bacterial group 2. Only compound bacterial groups 2 to 4 were higher than the sum of the concentrations measured in each single-bacterial group. Among them, compound bacterial group 4 unexpectedly achieved a significantly increased IFN-γ concentration, which was significantly better than that in compound bacterial groups 2 and 3. This indicates that when Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 are used in combination at a bacterial ratio of 1:0.11 to 1:9, they can synergistically induce Th1 cells to secrete IFN-γ, especially at a bacterial ratio of 1:9.

[0082] Based on the above, the applicant further took the supernatant from the pathological control group, each single bacterial group, and the compound bacterial group 4 for analysis in item C below.

[0083] C. Determination of IL-10 and IL-13 concentrations:

[0084] The concentrations (pg / mL) of IL-10 and IL-13 in the supernatant of each group were determined using the IL-10 ELISA kit (ThermoFisher, catalog number BMS215-2) and the IL-13 ELISA kit (ThermoFisher, catalog number BMS231-3) and in accordance with the manufacturer’s instructions.

[0085] Next, the obtained experimental data were analyzed according to the method described in item 1 of the "General Experimental Methods" section above. The results are shown in... Figure 5 and Figure 6 middle.

[0086] Figure 5 This shows the IL-10 concentrations of PBMCs in each group after treatment with different lactic acid bacteria strains. Figure 5 It is evident that, compared with the pathological control group, the IL-10 concentrations in each single bacterial group and the complex bacterial group 4 were significantly increased. In particular, the IL-10 concentration in the complex bacterial group 4 was significantly higher than that in each single bacterial group. This indicates that when Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 are used in combination at a bacterial count ratio of 1:9, they can synergistically induce Th1 cells to secrete IL-10.

[0087] Figure 6 This shows the IL-13 concentrations of PBMCs in each group after treatment with different strains of lactic acid bacteria. Figure 6 It is evident that, compared with the pathological control group, the IL-13 concentrations in each single bacterial group and the complex bacterial group 4 were significantly reduced. In particular, the IL-13 concentration in the complex bacterial group 4 was significantly lower than that in each single bacterial group. This indicates that when Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 are used in combination at a bacterial count ratio of 1:9, they can synergistically inhibit the secretion of IL-13 by Th2 cells.

[0088] Based on the above experimental results, it can be concluded that using Lactobacillus salivarius AP-32 and Bifidobacterium animalis subsp. lactis CP-9 in a bacterial count ratio of 1:9 can synergistically improve the Th1 / Th2 imbalance, thereby significantly enhancing the therapeutic efficacy of atopic dermatitis.

[0089] All patents and documents cited in this specification are incorporated herein by reference in their entirety. In the event of any conflict, the detailed description of this application (including limitations) shall prevail.

[0090] Although the invention has been described with reference to the specific examples described above, it is apparent that many modifications and variations can be made without departing from the scope and spirit of the invention. Therefore, it is intended that the invention be limited only to those shown in the appended claims.

[0091] Information on the Preservation of Biological Materials

[0092] Accession number: CCTCC NO: M 2011127

[0093] Classification and naming: Lactobacillus salivarius subsp. salicinius AP-32

[0094] Date of deposit: April 10, 2011

[0095] Depository Institution: China Center for Type Culture Collection

[0096] Address of the depository: Wuhan, China

[0097] Accession number: CCTCC NO: M 2014588

[0098] Classification and naming: Bifidobacterium animalis subsp. lactis CP-9

[0099] Date of deposit: November 24, 2014

[0100] Depository Institution: China Center for Type Culture Collection

[0101] Address of the depository: Wuhan, China.

Claims

1. The use of lactic acid bacteria in the preparation of pharmaceutical compositions for the treatment of atopic dermatitis, characterized in that: The lactic acid bacteria consisted of Lactobacillus salivarius subsp. salicinius AP-32 with accession number CCTCC NO: M 2011127 and Bifidobacterium animalis subsp. lactis CP-9 with accession number CCTCC NO: M 2014588. The ratio of Lactobacillus salivarius subsp. salicinius AP-32 to Bifidobacterium animalis subsp. lactis CP-9 fell within the range of 1:0.11 to 1:

9.

2. The use according to claim 1, characterized in that: The bacterial count ratio is 1:

9.

3. The use according to claim 1, characterized in that: The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

4. The use according to claim 1, characterized in that: The pharmaceutical composition is in a dosage form for oral or topical administration.

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