FPR2 agonist containing outer membrane vesicles derived from lactic acid bacterium

Abstract

The present invention provides an FPR2 agonist. More specifically, the FPR2 agonist contains, as an active ingredient, outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of lactic acid bacteria belonging to the genera Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus and Lactococcus and Streptococcus salivarius subsp. thermophilus.

Description

FPR2 agonist containing outer membrane vesicles derived from lactic acid bacteria

[0001] The present invention relates to an invention characterized by using outer membrane vesicles derived from lactic acid bacteria to activate FPR2.

[0002] Outer membrane vesicles are lipid bilayer-enclosed substances released from various cells. They can cross the blood-brain barrier (Non-Patent Documents 5 and 6). They remain stable even after being taken up by host cells (Non-Patent Document 7). Outer membrane vesicles with these characteristics are expected to be applied to drug delivery systems (Non-Patent Documents 6 and 7). Bacteria also release outer membrane vesicles. Outer membrane vesicles derived from lactic acid bacteria have physiological activities such as suppressing the secretion of inflammatory mediators (e.g., IL-6), inducing differentiation into M2-type macrophages, suppressing inflammatory responses by macrophages, and enhancing the host immune response to vancomycin-resistant enterococci (Patent Documents 1 to 6 and Non-Patent Documents 1 to 2). It has also been reported that outer membrane vesicles derived from Lactiprantibacillus plantarum induce IL-10 (Non-Patent Document 14), and that outer membrane vesicles derived from Lacticaseibacillus paracasei (formerly classified as Lactobacillus paracasei) reduce inflammatory responses in the intestinal tract (Non-Patent Document 15).

[0003] Formyl peptide receptors (FPRs) are receptors for formylmethionyl peptides, known as chemotactic factors. Humans have three FPR families (FPR1, FPR2, and FPR3). Various FPR ligands exist, but the selectivity for each FPR varies depending on the ligand (Non-Patent Document 3). Various substances are known to act as agonists or antagonists of FPR2 (Non-Patent Document 3). FPR2 regulates both innate immunity, mediated by phagocytes, and adaptive immunity, mediated by B cells and other cells (Patent Document 7). FPR2 plays an important role in host defense and inflammatory responses and is recognized as a therapeutic target for inflammatory diseases (Non-Patent Document 4). FPR2 agonists are known to treat diseases associated with FPR2, such as inflammatory diseases (Patent Document 7). FPR2 (FPRL-1) is also a functional receptor for amyloid beta-42 (Aβ42), which is important in the pathophysiology of neurotoxicity associated with Alzheimer's disease (Non-Patent Document 8). It has been suggested that the FPR2-binding peptide Humanin (HN) may exert neuroprotective effects by competitively inhibiting FPR2's access to Aβ42 (Non-Patent Document 8). Humanin (HN) acts more strongly on FPR2 than on FPR1 (Non-Patent Document 9). FPR2 is expressed in colonic epithelial cells and mediates N-formyl peptide-dependent epithelial cell proliferation and regeneration. FPR2 also plays an important role in maintaining colonic homeostasis, inflammation, and epithelial repair (Non-Patent Document 10). The therapeutic effect of FPR2 (FPRL1) ligands on ulcerative colitis suggests the utility of FPR2 in the development of therapeutics for chronic inflammatory bowel disease (Non-Patent Document 11). Agonists of FPR2 have also been shown to be beneficial in preclinical models of chronic inflammatory human diseases, including infectious diseases, psoriasis, dermatitis, inflammatory bowel syndrome, Crohn's disease, ocular inflammation, sepsis, pain, metabolic / diabetic, cancer, COPD, asthma and allergic diseases, cystic fibrosis, acute lung injury and fibrosis, rheumatoid arthritis and other joint diseases, Alzheimer's disease, renal fibrosis, and organ transplantation (Patent Document 7 and Non-Patent Documents 12 and 13).

[0004] Special table No. 2021-516054 Publication Special table No. 2021-513336 Publication Special table No. 2021-502970 Publication Special table No. 2021-501136 Publication Special table No. 2018-511583 Publication Special table No. 2013-530216 Publication Special table No. 2021-516240 Publication

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[0006] Substances that activate FPR2 are thought to be useful for treating diseases associated with FPR2. Outer membrane vesicles have properties (such as excellent stability) that are advantageous for such treatments. However, no outer membrane vesicles that activate FPR2 are known. Therefore, we set the goal of providing outer membrane vesicles that can be activated by FPR2.

[0007] As a result of extensive research into this issue, the present inventors have found that outer membrane vesicles derived from specific types of lactic acid bacteria activate FPR2, and the present invention was made based on this finding.

[0008] That is, the present invention relates to the following [1] to

[36] : [1] An FPR2 agonist containing outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.[2] The lactic acid bacteria are any of the following: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Lactococcus lactis subsp. lactis, The FPR2 agonist according to [1] above, which is at least one selected from the group consisting of Streptococcus salivarius subsp. thermophilus, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticaseibacillus casei, and Lactiplantibacillus pentosus. [3] The FPR2 agonist according to [1] or [2] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.[4] A food or beverage for activating FPR2, containing as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus. [5] A pharmaceutical composition for treating a disease associated with FPR2, comprising, as an active ingredient, outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus. [6] A cosmetic composition for activating FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.[7] A consumer product composition for activating FPR2, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus. [8] The food or beverage product according to [4], the pharmaceutical composition according to [5], the cosmetic composition according to [6], or the consumer product composition according to [7], wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity. [9] Lacticaseibacillus paracasei strain CP3526, having the accession number NITE BP-03625.

[10] A food or beverage for inducing IL-10 through FPR2 activation, comprising, as an active ingredient, outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genera Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[11] A pharmaceutical composition for treating a disease associated with IL-10 through IL-10 induction by FPR2 activator, comprising, as an active ingredient, outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[12] A cosmetic composition for inducing IL-10 through FPR2 activation, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[13] A consumer product composition for inducing IL-10 through FPR2 activation, comprising, as an active ingredient, outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[14] The food or drink according to

[10] , the pharmaceutical composition according to

[11] , the cosmetic composition according to

[12] , or the consumer product composition according to

[13] , wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[15] A food or drink for protecting the skin from ultraviolet rays, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[16] A pharmaceutical composition for protecting skin from ultraviolet rays, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[17] A cosmetic composition for protecting skin from ultraviolet rays, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[18] A consumer product composition for protecting skin from ultraviolet rays, comprising as an active ingredient outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[19] The food or beverage according to

[15] , the pharmaceutical composition according to

[16] , the cosmetic composition according to

[17] , or the consumer product composition according to

[18] , which protects skin from ultraviolet rays by FPR2 activation.

[20] The food or drink according to

[15] , the pharmaceutical composition according to

[16] , the cosmetic composition according to

[17] , or the consumer product composition according to

[18] , wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[21] A food or drink for protecting neurons from neurotoxicity, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacteria selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[22] A pharmaceutical composition for protecting neurons from neurotoxicity, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[23] A cosmetic composition for protecting nerve cells from neurotoxicity, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[24] A consumer product composition for protecting nerve cells from neurotoxicity, comprising as an active ingredient outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of Lactobacillus, Lacticaseibacillus, Lactiplantibacillus, Ligilactobacillus, Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[25] The food or drink according to

[21] , the pharmaceutical composition according to

[22] , the cosmetic composition according to

[23] , or the consumer product composition according to

[24] , which protects neurons from neuronal toxicity by activating FPR2.

[26] The food or drink according to

[21] , the pharmaceutical composition according to

[22] , the cosmetic composition according to

[23] , or the consumer product composition according to

[24] , wherein the neuronal toxicity is neuronal toxicity caused by amyloid beta.

[27] The food or drink according to

[21] , the pharmaceutical composition according to

[22] , the cosmetic composition according to

[23] , or the consumer product composition according to

[24] , wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[28] A method for activating FPR2 in a subject, the method comprising the step of administering to the subject outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[29] A method for treating a disease associated with FPR2 in a subject, comprising the step of administering to the subject outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[30] A method for inducing IL-10 in a subject by activating FPR2, comprising the step of administering to the subject outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[31] A method for protecting the skin of a subject from ultraviolet rays, comprising the step of administering to the subject outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[32] The method according to

[31] , wherein the skin is protected from ultraviolet rays by activating FPR2.

[33] A method for protecting neurons in a subject from neuronal toxicity, comprising the step of administering to the subject outer membrane vesicles derived from at least one type of lactic acid bacterium selected from the group consisting of the genus Lactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Ligilactobacillus, the genus Lactococcus, and Streptococcus salivarius subsp. thermophilus.

[34] The method according to

[33] , wherein neurons are protected from neuronal toxicity by activating FPR2.

[35] The lactic acid bacterium is selected from the group consisting of Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Ligilactobacillus salivarius, Lactococcus lactis subsp. lactis, The method according to any one of

[28] to

[34] above, wherein the lactic acid bacterium is at least one selected from the group consisting of Streptococcus salivarius subsp. thermophilus, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticaseibacillus casei, and Lactiplantibacillus pentosus.

[36] The method according to any one of

[28] to

[35] above, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonist activity.

[0009] As shown in the Examples below, outer membrane vesicles derived from specific lactic acid bacteria activate FPR2. Furthermore, as shown in the Examples below, outer membrane vesicles derived from specific lactic acid bacteria induce IL-10 through activation of FPR2, protect skin from ultraviolet rays, and protect neurons from neuronal toxicity. The present invention, which uses these outer membrane vesicles as an active ingredient, is useful for treating diseases associated with FPR2.

[0010] Figure 1 shows the FPR2 agonistic activity of 26 types of bacterial-derived outer membrane vesicles. Figure 2 shows the FPR1 agonistic activity of 26 types of bacterial-derived outer membrane vesicles. Figure 3 shows the effect of known FPR2 agonists on IL-10 production by M2 macrophages. Figure 4 shows the IL-10-inducing activity of 17 types of bacterial-derived outer membrane vesicles. Figure 5 shows the effect of an FPR2 antagonist on the IL-10-inducing ability of outer membrane vesicles with FPR2 agonistic activity. Figure 6 shows the effect of outer membrane vesicles with FPR2 agonistic activity on protecting skin from UV rays. Figure 7 shows the effect of outer membrane vesicles with FPR2 agonistic activity on protecting neurons from neuronal toxicity.

[0011] [Lactic Acid Bacteria] The outer membrane vesicles used in the present invention are derived from the following lactic acid bacteria (1) to (6): (1) Lactobacillus genus (2) Lacticaseibacillus genus (3) Lactiplantibacillus genus (4) Ligilactobacillus genus (5) Lactococcus genus (6) Streptococcus salivarius subsp. thermophilus The classification of lactic acid bacteria in this specification follows the reclassification published in 2020. Details of the reclassification are described in Int J Syst Evol Microbiol. 2020 Apr; 70(4): 2782-2858 and on the website of the Japanese Society of Enterobacteriaceae (URL: https: / / bifidus-fund.jp / keyword / kw054.shtml).

[0012] Any known lactic acid bacteria strain belonging to any of the above (1) to (6) and secreting outer membrane vesicles can be used in the present invention without any particular limitations. Since outer membrane vesicles exert their effects when taken orally or ingested, strains whose safety in animals has been confirmed are preferred.

[0013] Preferred lactic acid bacteria are those of the genus (1) Lactobacillus, (2) Lacticaseibacillus, (3) Lactipranchibacillus, (4) Rizilactobacillus, or (5) Lactococcus.

[0014] More preferred lactic acid bacteria are those of the genus (1) Lactobacillus, (2) Lacticaseibacillus, (3) Lactipranchibacillus, or (4) Rizilactobacillus.

[0015] More preferred lactic acid bacteria are those of the genus (1) Lactobacillus, (2) Lacticaseibacillus, or (4) Rizilactobacillus.

[0016] Particularly preferred lactic acid bacteria are those of the genus (1) Lactobacillus or (2) Lacticaseibacillus.

[0017] (1) Lactobacillus genus lactic acid bacteria Examples of Lactobacillus genus lactic acid bacteria include the following: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus amylovorus, Lactobacillus johnsonii

[0018] Preferred lactobacillus lactic acid bacteria are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus delbrueckii or Lactobacillus gasseri. In one aspect of the present invention, preferred lactobacillus lactic acid bacteria are Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus or Lactobacillus johnsonii. In another aspect of the present invention, preferred lactobacillus lactic acid bacteria are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus or Lactobacillus johnsonii.

[0019] More preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, or Lactobacillus acidophilus. In one embodiment of the present invention, more preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, or Lactobacillus johnsonii.

[0020] Even more preferred lactic acid bacteria of the genus Lactobacillus are Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus gasseri or Lactobacillus johnsonii.

[0021] Lactobacillus delbrueckii subsp. bulgaricus may be any available strain and may include the type strain Lactobacillus delbrueckii subsp. bulgaricus T. Lactobacillus helveticus may be any available strain and may include the type strain Lactobacillus helveticus T. Lactobacillus acidophilus may be any available strain and may include the type strain Lactobacillus acidophilus T. Lactobacillus acidophilus may include other strains, such as the JCM1023 strain and the CP1613 strain. The Lactobacillus acidophilus CP1613 strain is a species of lactic acid bacterium belonging to the Lactobacillus acidophilus genus, and was internationally deposited on January 18, 2013, at the Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818), an international depository institution under the provisions of the Budapest Treaty, under accession number NITE BP-1513. Any available strain of Lactobacillus gasseri may be used, and may include the type strain Lactobacillus gasseri T. Other strains of Lactobacillus gasseri may also be used, such as the CP2305 strain. Lactobacillus gasseri CP2305 strain is a species of lactic acid bacterium belonging to the Lactobacillus gasseri genus and has been internationally deposited under the Budapest Treaty at the International Patent Organism Depositary of the National Institute of Technology and Evaluation (Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818) under accession number FERM BP-11331 (deposit date: September 11, 2007). Lactobacillus amylovorus may be any available strain, including the type strain Lactobacillus amylovorus T. Lactobacillus crispatus may be any available strain, including the type strain Lactobacillus crispatus T. Lactobacillus johnsonii may be any available strain, including the type strain Lactobacillus johnsonii T.

[0022] (2) Lactic acid bacteria of the genus Lacticaseibacillus Examples of lactic acid bacteria of the genus Lacticaseibacillus include the following: Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lacticaseibacillus casei, and Lacticaseibacillus paracasei subsp. paracasei.

[0023] Preferred lactic acid bacteria of the genus Lacticaseibacillus are Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, and Lacticaseibacillus casei.

[0024] More preferred lactic acid bacteria of the genus Lacticaceibacillus are Lacticaceibacillus paracasei or Lacticaceibacillus casei. Lacticaceibacillus rhamnosus may be any available strain, and may include the type strain Lacticaceibacillus rhamnosus T. Lacticaceibacillus paracasei may be any available strain, and may include the type strain Lacticaceibacillus paracasei T. Lacticaceibacillus paracasei may include other strains, such as the JCM1133 strain and the CP3526 strain. The Lacticaseibacillus paracasei CP3526 strain is a species of lactic acid bacterium belonging to the Lacticaseibacillus paracasei genus, and has been internationally deposited at the Patent Microorganisms Depositary of the National Institute of Technology and Evaluation (Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818), an international depository institution under the provisions of the Budapest Treaty, under accession number NITE BP-03625, dated March 18, 2022. The Lacticaseibacillus casei may be any available strain, and may include the type strain Lacticaseibacillus casei T.

[0025] In one embodiment of the present invention, the lactic acid bacteria of the genus Lacticaseibacillus may be a lactic acid bacterium other than Lacticaseibacillus paracasei.

[0026] (3) Lactic acid bacteria of the genus Lactiplantibacillus Examples of lactic acid bacteria of the genus Lactiplantibacillus include the following: Lactiplantibacillus plantarum Lactiplantibacillus pentosus Lactiplantibacillus paraplantarum

[0027] A preferred lactic acid bacterium of the genus Lactiprancibacillus is Lactiprancibacillus plantarum or Lactiprancibacillus pentosus.

[0028] A more preferred lactic acid bacterium of the genus Lactiprancylbacillus is Lactiprancylbacillus plantarum.

[0029] Lactipranctibacillus plantarum may be any available strain, and may include the type strain Lactipranctibacillus plantarum T. Lactipranctibacillus pentosus may be any available strain, and may include the type strain Lactipranctibacillus pentosus T.

[0030] In one embodiment of the present invention, the lactic acid bacterium of the genus Lactipranchibacillus may be a lactic acid bacterium other than Lactipranchibacillus plantarum.

[0031] (4) Lactic acid bacteria of the genus Ligilactobacillus Examples of lactic acid bacteria of the genus Ligilactobacillus include the following: Ligilactobacillus salivarius Ligilactobacillus acidipiscis

[0032] A preferred lactic acid bacterium of the genus Risilactobacillus is Risilactobacillus salivarius.

[0033] The Risilactobacillus salivarius may be any available strain, and may include the type strain Risilactobacillus salivarius T.

[0034] (5) Lactic acid bacteria of the genus Lactococcus Examples of lactic acid bacteria of the genus Lactococcus include the following: Lactococcus lactis subsp. lactis Lactococcus lactis subsp. cremoris Lactococcus lactis

[0035] A preferred lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis or Lactococcus lactis subsp. cremoris.

[0036] A more preferred lactic acid bacterium of the genus Lactococcus is Lactococcus lactis subsp. lactis.

[0037] Lactococcus lactis subsp. lactis may be any available strain, and may include the type strain Lactococcus lactis subsp. lactis T.

[0038] (6) Streptococcus salivarius subsp. thermophilus Streptococcus salivarius subsp. thermophilus may be any available strain, and may include the type strain Streptococcus salivarius subsp. thermophilus T.

[0039] Mutants or derivatives of the specific strains mentioned above may also be used as long as they secrete outer membrane vesicles.

[0040] Preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticazei Bacillus rhamnosus, Lacticazei Bacillus paracasei, Lactiprantibacillus plantarum, Lactobacillus salivarius, Lactococcus lactis subsp. lactis, and Streptococcus salivarius subsp. thermophilus. In one aspect of the present invention, preferred lactic acid bacteria are at least one species selected from the group consisting of the following options. Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticaseibacillus casei, and Lactipranchibacillus pentosus. In another aspect of the present invention, preferred lactic acid bacteria are at least one selected from the group consisting of the following options: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactiprancistibacillus plantarum, Lactobacillus salivarius, Lactococcus lactis subsp. lactis, Streptococcus salivarius subsp. thermophilus, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticaseibacillus casei, and Lactiprancistibacillus pentosus

[0041] More preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticazeibacillus rhamnosus, Lacticazeibacillus paracasei, Lactipranctibacillus plantarum, Lactobacillus salivarius, and Lactococcus lactis subsp. lactis. In one aspect of the present invention, more preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticazeibacillus casei, and Lactipranctibacillus pentosus. In another aspect of the present invention, more preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticazeibacillus casei, and Lactipranctibacillus pentosus. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactipranchibacillus plantarum, Lactobacillus salivarius, Lactococcus lactis subsp. lactis, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticaseibacillus casei, and Lactipranchibacillus pentosus

[0042] More preferred lactic acid bacteria are at least one species selected from the group consisting of the following: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticazeibacillus rhamnosus, Lacticazeibacillus paracasei, Lactipranctibacillus plantarum, and Rizilactobacillus salivarius. In one aspect of the present invention, more preferred lactic acid bacteria are at least one species selected from the group consisting of the following: Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticazeibacillus casei, and Lactipranctibacillus pentosus. In another aspect of the present invention, more preferred lactic acid bacteria are at least one species selected from the group consisting of the following: Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticazeibacillus casei, and Lactipranctibacillus pentosus. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactiprancistibacillus plantarum, Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, Lacticaseibacillus casei, and Lactiprancistibacillus pentosus

[0043] An even more preferred lactic acid bacterium is at least one selected from the group consisting of the following: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticazei Bacillus rhamnosus, Lacticazei Bacillus paracasei, and Lactobacillus salivarius. In one aspect of the present invention, an even more preferred lactic acid bacterium is at least one selected from the group consisting of the following: Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, and Lacticazei Bacillus casei. In another aspect of the present invention, an even more preferred lactic acid bacterium is at least one selected from the group consisting of the following: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticazei Bacillus paracasei, and Lacticazei Bacillus casei. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactobacillus salivarius, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, and Lacticaseibacillus casei

[0044] Particularly preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticazei Bacillus rhamnosus, and Lacticazei Bacillus paracasei. In one aspect of the present invention, particularly preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, and Lacticazei Bacillus casei. In another aspect of the present invention, particularly preferred lactic acid bacteria are at least one species selected from the group consisting of the following options: Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus, Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei, Lactobacillus gasseri, Lactobacillus amylovorus, Lactobacillus crispatus, Lactobacillus johnsonii, and Lacticaseibacillus casei

[0045] More particularly preferred lactic acid bacteria are at least one selected from the group consisting of the following: Lactobacillus acidophilus, Lacticaseibacillus paracasei, and Lactobacillus gasseri. Most preferred lactic acid bacteria are at least one selected from the group consisting of the following: Lactobacillus acidophilus, and Lacticaseibacillus paracasei.

[0046] A single type of lactic acid bacteria may be used, or multiple types may be used in combination. Therefore, outer membrane vesicles derived from two or more types of lactic acid bacteria can be used in the present invention.

[0047] [Lactic acid bacteria-derived outer membrane vesicles] Bacterial outer membrane vesicles are substances secreted by bacteria. They are spherical structures composed of the lactic acid bacteria's cell membrane (lipid bilayer), and their diameters are generally around 20 to 500 nm. The interior of these spherical structures may contain nucleic acids (DNA and RNA) and proteins (enzymes, etc.). Lactic acid bacteria-derived outer membrane vesicles may also contain membrane proteins present in the lipid bilayer membrane of lactic acid bacteria, such as peptidoglycan (PG) (Journal of the Japanese Society for Lactic Acid Bacteria, 27(1), 10-16, 2016, and Microbiol Spectr., 7(1)., 2019). Bacterial outer membrane vesicles are also called membrane vesicles, extracellular vesicles (EV), extracellular vesicles, microvesicles, membrane vesicles (MV), outer membrane vesicles (OMV), etc.

[0048] No special culture conditions are required for the secretion of outer membrane vesicles; they are released into the medium by culturing under conditions commonly used in lactic acid bacteria culture. The medium preferably contains a carbon source, a nitrogen source, inorganic salts, etc., and is capable of efficiently culturing lactic acid bacteria. The medium may be either a natural or synthetic medium, and a known medium suitable for the strain used can be appropriately selected. Examples of carbon sources include lactose, glucose, sucrose, fructose, galactose, blackstrap molasses, etc. Examples of nitrogen sources include organic nitrogen-containing substances such as casein hydrolysate, whey protein hydrolysate, and soy protein hydrolysate. Examples of inorganic salts include phosphate, sodium, potassium, and magnesium. Examples of media suitable for lactic acid bacteria include MRS medium, GLM medium, GAM medium, BL medium, Briggs Liver Broth, animal milk, skim milk, and dairy whey, with MRS medium and GLM medium being preferred. Furthermore, when outer membrane vesicles of lactic acid bacteria are used for food applications, a medium composed only of food materials and food additives can be used.

[0049] Lactic acid bacteria can be cultured under static or anaerobic conditions at a temperature of, for example, 20 to 50°C (preferably 30 to 40°C). The culture temperature can be adjusted using a thermostatic bath, a mantle heater, a jacket, or the like. Anaerobic conditions refer to a low-oxygen environment in which lactic acid bacteria can grow. Anaerobic conditions can be achieved using an anaerobic chamber, an anaerobic box, or a sealed container or bag containing an oxygen absorber. Examples of culture formats include static culture, agitation culture, and tank culture. The culture time is, for example, 3 to 96 hours, preferably 12 to 32 hours.

[0050] OMVs can be obtained from a lactic acid bacteria culture by appropriately combining one or more known separation and purification methods based on the mass and size of the OMVs. Examples of separation and purification methods include centrifugation, ultracentrifugation, and filtration. For example, OMVs can be obtained using the following procedure: A lactic acid bacteria culture is centrifuged (e.g., at a centrifugal force of 3,000 to 15,000 g, preferably 8,000 to 10,000 g) to precipitate the bacterial cells. The supernatant after centrifugation is filtered through a filter with a pore size that allows OMVs to pass through (e.g., 220 to 1,000 nm, preferably 450 nm (0.45 μm)). The filtrate containing the OMVs is subjected to ultracentrifugation (e.g., at a centrifugal force of 100,000 to 300,000 g, preferably 150,000 to 300,000 g) to obtain the OMVs as a precipitate. The precipitate may be further filtered.

[0051] [FPR2 (Formyl Peptide Receptor 2)] FPR2 (Formyl Peptide Receptor 2) is a receptor for peptides containing N-formylmethionine (e.g., f-Met-Leu-Phe (FMLP)) and is a G protein-coupled receptor. FPR2 is a member of the human FPR family (FPR1, FPR2, and FPR3). FPR2 is also called FPRL-1 or FPRL1. FPR1 is called the formyl peptide receptor, and FPR2 and FPR3 are called formyl peptide-like receptors. FPR2 shares higher sequence homology with FPR3 than with FPR1 (Pharmacol Rev 61:119-161, 2009).

[0052] [FPR2 Agonist] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to activate FPR2 (activate it by binding to FPR2). Therefore, one aspect of the present invention is an FPR2 agonist that contains outer membrane vesicles derived from specific lactic acid bacteria as an active ingredient. Furthermore, the use of outer membrane vesicles derived from specific lactic acid bacteria to prepare an FPR2 agonist is also an aspect of the present invention. Although substances that have agonist activity against FPR2, such as low-molecular-weight compounds and peptides, there have been no reports to date that outer membrane vesicles derived from lactic acid bacteria activate FPR2. The present invention uses outer membrane vesicles derived from specific lactic acid bacteria that can activate FPR2 as an FPR2 agonist. The lactic acid bacteria-derived outer membrane vesicles used as the active ingredient of the FPR2 agonist in this invention are known to be very stable and can remain in the cells for a long time without being completely decomposed, and therefore, unlike conventional substances that have agonistic activity against FPR2, they are thought to have both FPR2 agonist function and the ability to be delivered stably to the body. Furthermore, lactic acid bacteria-derived outer membrane vesicles can be easily obtained by culturing lactic acid bacteria as described above. FPR2 agonists have been shown to be beneficial in resolving chronic inflammation and in preclinical models of chronic inflammatory human diseases, including infectious diseases, psoriasis, dermatitis, inflammatory bowel syndrome, Crohn's disease, ocular inflammation, sepsis, pain, metabolism / diabetes, cancer, COPD, asthma and allergic diseases, cystic fibrosis, acute lung injury and fibrosis, rheumatoid arthritis and other joint diseases, Alzheimer's disease, renal fibrosis, and organ transplantation (Patent Document 7 and Non-Patent Documents 12 and 13). Therefore, it is believed that outer membrane vesicles derived from specific lactic acid bacteria of the present invention are useful for treating diseases associated with FPR2.

[0053] FPR2 agonist activity can be measured using a known assay system capable of measuring ligand-induced activation of G protein-coupled receptors (GPCRs) (e.g., the reporter assay described in International Publication No. WO 2020 / 026979). This reporter assay is offered as a contract service (e.g., Tanso Biosciences GPCR contract testing, a subsidiary of Cosmo Bio Co., Ltd.).

[0054] The outer membrane vesicles of lactic acid bacteria according to the present invention preferably selectively agonize FPR2. "Selectively agonizing FPR2" means having agonistic activity against FPR2 but not against FPR1. Because FPR1 is associated with the induction of inflammatory responses (Non-Patent Document 3), FPR2 agonists that "selectively agonize FPR2" can treat diseases associated with FPR2 without inducing inflammatory responses.

[0055] [Diseases associated with FPR2] FPR2 agonists can be used to treat diseases associated with FPR2. "Diseases associated with FPR2" include not only those that are the target of pharmaceuticals, but also those that are the target of functional foods, beverages, and functional cosmetics. "Treatment" includes not only actions aimed at treatment and prevention (use of FPR2 agonists as pharmaceuticals or quasi-drugs), but also actions aimed at health promotion (use of FPR2 agonists as functional foods, beverages, and functional cosmetics). FPR2 agonists can be used for both therapeutic (medical) and non-therapeutic (non-medical) purposes.

[0056] Examples of "diseases associated with FPR2" include inflammatory diseases, heart disease, chronic airway disease, cancer, sepsis, allergic conditions, circulatory disorders, neuroinflammation, neuropathy, pain, prion diseases, amyloidosis, immune disorders, Behcet's disease, Sweet's disease, systemic lupus erythematosus (SLE), Wegener's granulomatosis, viral infections, diabetes, bacterial infections, physical injuries, physical injuries including radiation, vasoconstriction, anaphylactic reactions, allergic reactions, rhinitis, shock (endotoxic, hemorrhagic, traumatic, visceral ischemic, and circulatory shock), rheumatoid arthritis, and pain. Colds, benign prostatic hyperplasia, myocardial ischemia, myocardial infarction, heart failure, brain injury, lung disease, COPD, COAD, COLD, acute lung injury, acute respiratory distress syndrome, chronic bronchitis, emphysema, asthma (allergic and non-allergic asthma), cystic fibrosis, renal fibrosis, kidney damage, glomerular disease, ulcerative colitis, IBD, Crohn's disease, periodontitis, pain, Alzheimer's disease, AIDS, uveitis, glaucoma, conjunctivitis, Sjogren's syndrome, atherosclerosis, neuroinflammation including multiple sclerosis, stroke , skin diseases (rosacea, sunburn, psoriasis, hot flashes, flushing and redness associated with hot flashes, erythema associated with hot flashes, photoaging, seborrheic dermatitis, acne, allergic dermatitis, vascular ectasia, rhinophyma, skin rash, skin erythema, hyperactivity of the skin with dilated skin blood vessels, Lyell's syndrome, Stevens-Johnson syndrome, localized itching and discomfort associated with hemorrhoids, hemorrhoids, erythema multiforme minor, erythema multiforme major, erythema nodosum, eye swelling, urticaria, pruritus, purpura, varicose veins, contact dermatitis, atopic dermatitis, Skin eczema, nummular dermatitis, generalized exfoliative dermatitis, stasis dermatitis, simple chronic lichen planus, perioral dermatitis, pseudofolliculitis barbae, granuloma annulare, actinic keratosis, basal cell carcinoma, squamous cell carcinoma, eczema, skin wound healing, hypertrophic scars, keloids, burns, actinic keratosis, melanoma, viral warts, photoaging, photodamage, melasma, post-inflammatory hyperpigmentation, other hyperpigmentation disorders, alopecia), eye inflammation, enteritis, inflammatory bowel syndrome, dermatitis, tonsillitis, laryngitis, pharyngitis, sinusitis, esophagitis, gastritis, etc.

[0057] The above diseases can be classified, for example, as follows: (1) inflammatory diseases (e.g., tonsillitis, laryngitis, pharyngitis, eye inflammation, rhinitis, sinusitis, periodontitis, conjunctivitis, neuroinflammation, chronic bronchitis, Behcet's disease, etc.) (2) digestive diseases (e.g., esophagitis, gastritis, enteritis, inflammatory bowel syndrome, ulcerative colitis, etc.) (3) skin diseases (e.g., dermatitis (including allergic dermatitis, atopic dermatitis, and seborrheic dermatitis), acne, rash, sunburn, hot flashes, eczema, photoaging, itching and discomfort, perioral dermatitis, hyperpigmentation disorder, alopecia, etc.) (4) brain diseases (e.g., dementia (including alcoholic dementia), migraine, Alzheimer's disease (including early symptoms such as forgetfulness), prion diseases, etc.) (5) Other diseases (respiratory diseases (e.g., asthma, chronic obstructive pulmonary disease (COPD), emphysema, etc.), cardiovascular diseases (e.g., atherosclerosis, heart disease, circulatory disorders, myocardial ischemia, myocardial infarction, heart failure, etc.), joint diseases (e.g., rheumatoid arthritis, etc.), neurological diseases (e.g., neuritis, neuropathy, pain, etc.), metabolic diseases (e.g., diabetes, gout, kidney disorders, etc.), infectious diseases (e.g., bacterial infection, viral infection, sepsis caused by infection, etc.), immune disorders (e.g., allergic symptoms, AIDS, etc.), others (e.g., physical trauma, cancer, Crohn's disease, etc.)

[0058] FPR2 is known to be involved in brain neurotoxicity and in the homeostasis, inflammation, proliferation, and regeneration of intestinal epithelial cells (see Non-Patent Documents 8 to 11, cited above). Epithelial cells are also present in the skin. Furthermore, outer membrane vesicles can cross the blood-brain barrier (see Non-Patent Documents 5 and 6). Therefore, by binding to and activating FPR2, FPR2 agonists can be suitably used to treat brain diseases (e.g., Alzheimer's disease (including early symptoms such as forgetfulness), dementia (including alcoholic dementia), and migraine), digestive diseases (e.g., esophagitis, gastritis, enteritis, inflammatory bowel syndrome, and ulcerative colitis), and skin diseases (e.g., dermatitis (including allergic dermatitis, atopic dermatitis, and seborrheic dermatitis), acne, rash, sunburn, hot flashes, eczema, photoaging, itching and discomfort, perioral dermatitis, hyperpigmentation disorders, and alopecia).

[0059] The content of lactic acid bacteria-derived outer membrane vesicles in the FPR2 agonist can be appropriately determined depending on the condition and symptoms of the subject to whom the agonist is to be administered. The content (number) of outer membrane vesicles per mL of FPR2 agonist is, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12 cells / mL, more preferably 2 x 10 5 ~2x10 10 The FPR2 agonist concentration is 1 / mL. The FPR2 agonist can be applied to any animal that expresses FPR2. The target of application is preferably a mammal (human or non-human mammal (e.g., horse or cow)), more preferably a human. Furthermore, the target of application can be any gender or age.

[0060] [IL-10 Induction] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to induce the production and secretion of IL-10 (hereinafter also referred to as "IL-10 induction"). This embodiment can be understood as an IL-10 inducer containing outer membrane vesicles of lactic acid bacteria of the present invention as an active ingredient. In one embodiment of the present invention, the outer membrane vesicles induce IL-10 via FPR2 activation. This embodiment can be understood as an IL-10 inducer through FPR2 activation, containing outer membrane vesicles of lactic acid bacteria of the present invention as an active ingredient. IL-10 is an anti-inflammatory cytokine produced and secreted by immune cells such as macrophages, helper T cells, and regulatory T cells. IL-10 inducers (including embodiments in which IL-10 is induced via FPR2 activation; the same applies below) can be used to treat diseases associated with IL-10. "Diseases associated with IL-10" include not only those that are targets of pharmaceuticals but also those that are targets of functional foods, beverages, and functional cosmetics. "Treatment" includes not only actions aimed at treatment and prevention (use of IL-10 inducers as pharmaceuticals or quasi-drugs) but also actions aimed at health promotion (use of IL-10 inducers as functional foods and beverages or functional cosmetics). IL-10 inducers can be used for both therapeutic (medical) and non-therapeutic (non-medical) purposes. Examples of "diseases associated with IL-10" include the aforementioned (1) inflammatory diseases, (2) digestive diseases, (3) skin diseases, (4) brain diseases, and (5) other diseases. The content of lactic acid bacteria-derived outer membrane vesicles in an IL-10 inducer can be appropriately determined depending on the condition and symptoms of the subject to whom the IL-10 inducer is to be administered. The content (number) of outer membrane vesicles per mL of IL-10 inducer can be, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12 cells / mL, more preferably 2 x 10 5 ~2x10 10 The IL-10 inducer can be applied to any animal. The target animals are preferably mammals (humans and non-human mammals (e.g., horses and cows)), more preferably humans. Furthermore, the target animals can be any gender or age.

[0061] [Protection of Skin from UV Rays] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to protect skin from UV rays. This embodiment can be understood as a UV skin protectant containing outer membrane vesicles of lactic acid bacteria according to the present invention as an active ingredient. In one embodiment of the present invention, the outer membrane vesicles protect skin from UV rays via FPR2 activation. This embodiment can be understood as a UV skin protectant containing outer membrane vesicles of lactic acid bacteria according to the present invention as an active ingredient, via FPR2 activation. UV skin protectants (including those that protect skin via FPR2 activation; the same applies below) can be used to treat UV-associated skin diseases. "UV-associated skin diseases" include not only those that are the target of pharmaceuticals, but also those that are the target of functional foods, beverages, and functional cosmetics. "Treatment" includes not only actions aimed at treatment and prevention (use of skin protectants as pharmaceuticals or quasi-drugs) but also actions aimed at health promotion (use of skin protectants as functional foods, beverages, and functional cosmetics). The skin protective agent can be used for both therapeutic (medical) and non-therapeutic (non-medical) purposes. Examples of "skin diseases associated with ultraviolet rays" include the skin diseases (3) mentioned above. The content of lactic acid bacteria-derived outer membrane vesicles in the skin protective agent can be appropriately set depending on the condition and symptoms of the subject to whom the skin protective agent is applied. The content (number) of outer membrane vesicles per mL of the skin protective agent can be, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12 cells / mL, more preferably 2 x 10 5 ~2x10 10 The skin protective agent can be applied to any animal. The subject of application is preferably a mammal (human or non-human mammal (e.g., horse or cow)), more preferably a human. Furthermore, the gender and age of the subject are not important.

[0062] [Protection of Neurons from Neurotoxicity] In the present invention, outer membrane vesicles derived from specific lactic acid bacteria are used to protect neurons from neurotoxicity. This embodiment can be understood as a neuroprotective agent against neurotoxicity, containing outer membrane vesicles of the lactic acid bacteria of the present invention as an active ingredient. In one embodiment of the present invention, the outer membrane vesicles protect neurons from neurotoxicity via FPR2 activation. This embodiment can be understood as a neuroprotective agent against neurotoxicity caused by FPR2 activator, containing outer membrane vesicles of the lactic acid bacteria of the present invention as an active ingredient. Examples of neurotoxicity include those caused by endogenous toxins (e.g., amyloid beta and oxygen radicals) and exogenous toxins (e.g., ethanol). Neuroprotective agents against neurotoxicity (including those that protect neurons via FPR2 activation; the same applies below) can be used to treat diseases associated with neurotoxicity. "Diseases associated with neurotoxicity" include not only those that are the target of pharmaceuticals, but also those that are the target of functional foods, beverages, and functional cosmetics. "Treatment" includes not only actions aimed at treatment and prevention (use of neuronal protective agents as pharmaceuticals or quasi-drugs) but also actions aimed at health promotion (use of neuronal protective agents as functional foods and beverages or functional cosmetics). Neuronal protective agents can be used for both therapeutic (medical) and non-therapeutic (non-medical) purposes. Examples of "diseases associated with neuronal toxicity" include the aforementioned (4) brain diseases. The content of lactic acid bacteria-derived outer membrane vesicles in the neuronal protective agent can be appropriately set depending on the condition and symptoms of the subject to whom the neuronal protective agent is applied. The content (number) of outer membrane vesicles per mL of neuronal protective agent can be, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12 cells / mL, more preferably 2 x 10 5 ~2x10 10 The neuroprotective agent can be applied to animals without any restrictions. The target of application is preferably mammals (humans and non-human mammals (e.g., horses and cows)), more preferably humans. Furthermore, the target of application can be any gender or age.

[0063] [Optional Ingredients] The FPR2 agonist may contain one or more optional ingredients to the extent that the action of the active ingredient is not impaired. The optional ingredients can be selected appropriately depending on the mode of use of the FPR2 agonist, etc. The optional ingredients may be additives used in medicines, cosmetics, foods and beverages, etc. Examples of additives include oils and fats (vegetable oils (soybean oil, corn oil, safflower oil, olive oil, etc.) and animal fats (beef tallow, sardine oil, etc.)); herbal medicines (royal jelly, ginseng, etc.); amino acids (glutamine, cysteine, leucine, arginine, etc.); polyhydric alcohols (ethylene glycol, polyethylene glycol, propylene glycol, glycerin, sugar alcohols (sorbitol, erythritol, xylitol, maltitol, mannitol, etc.)); natural polymers (gum arabic, agar, water-soluble corn fiber, gelatin, xanthan gum, etc.)); Examples of suitable additives include: gluten, gluten hydrolysates, lecithin, starch, dextrin, etc.); vitamins (vitamin C, B vitamins, etc.); minerals (calcium, magnesium, zinc, iron, etc.); dietary fiber (mannan, pectin, hemicellulose, etc.); surfactants (glycerin fatty acid esters, sorbitan fatty acid esters, etc.); purified water; excipients (glucose, corn starch, lactose, dextrin, etc.); stabilizers; pH adjusters; antioxidants; sweeteners; flavoring components; acidulants; coloring agents, and flavorings. Furthermore, the FPR2 agonist may optionally contain functional ingredients other than the active ingredient (outer membrane vesicles derived from lactic acid bacteria) (e.g., taurine, glutathione, carnitine, creatine, coenzyme Q, glucuronic acid, glucuronolactone, chili pepper extract, ginger extract, cacao extract, guarana extract, Garcinia cambogia extract, theanine, γ-aminobutyric acid, capsaicin, capsiate, various organic acids, flavonoids, polyphenols, catechins, xanthine derivatives, indigestible oligosaccharides such as fructooligosaccharides, polyvinylpyrrolidone, etc.). The optional ingredients are known substances and are readily available on the market or can be prepared. A single optional ingredient may be used, or multiple optional ingredients may be used in combination. The content of the optional ingredient can be appropriately determined depending on the purpose of the formulation. The above description of the optional ingredients also applies to the IL-10 inducer, skin protectant, and neuroprotectant according to the present invention.

[0064] [Pharmaceutical Compositions] The outer membrane vesicles derived from specific lactic acid bacteria of the present invention can be used as pharmaceuticals or quasi-drugs. Accordingly, one aspect of the present invention is a pharmaceutical composition containing outer membrane vesicles derived from lactic acid bacteria as an active ingredient. Furthermore, the use of outer membrane vesicles of lactic acid bacteria for preparing a pharmaceutical composition is also another aspect of the present invention. The pharmaceutical composition can be used to treat the aforementioned "FPR2-associated diseases." The pharmaceutical composition can be used as an active ingredient in a method for treating an "FPR2-associated disease," which comprises administering an effective amount to a subject suffering from the aforementioned "FPR2-associated disease." The dosage form of the pharmaceutical composition is not particularly limited. Examples of dosage forms include oral preparations (tablets, capsules, granules, powders, dustings, syrups, dry syrups, liquids, suspensions, inhalants, etc.); enteral preparations (suppositories, etc.); infusions; injections; and topical preparations (aerosols, liquids, emulsions, creams, ointments, gels, lotions, poultices, tapes, etc.). Among these, oral preparations and topical preparations are preferred. Liquid preparations such as solutions and suspensions may be in a form that can be dissolved or suspended in water or other suitable medium immediately before administration. Tablets, granules, etc. may be coated on the surface by a known method. The pharmaceutical composition may be a sustained-release preparation, delayed-release preparation, or immediate-release preparation prepared according to a known release control technology.

[0065] The pharmaceutical composition may optionally contain pharmaceutically acceptable carriers and additives, such as water, organic solvents, collagen, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymers, sodium alginate, water-soluble dextran, water-soluble dextrin, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, petrolatum, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, surfactants, and artificial cell structures (e.g., liposomes).

[0066] The pharmaceutical composition can be applied to any animal expressing FPR2 without limitation. The target animal is preferably a mammal (human or non-human mammal (e.g., horse or cow)), more preferably a human. Furthermore, the target animal can be any gender or age.

[0067] The dosage of the pharmaceutical composition can be appropriately determined depending on the type of target disease, the age and weight of the subject, the route of administration, the number of administrations, the purpose of administration, etc. When administered orally, the number of outer membrane vesicles per kg of subject body weight is, for example, 2 x 10 6 ~2x10 14 pieces / kg, preferably 2x10 8 ~2x10 13 The dosage interval can be appropriately set depending on the type of target disease, etc., and may be once a day or divided into several doses. The above explanation regarding the pharmaceutical composition also applies to the aspects of the present invention relating to the treatment of "diseases associated with IL-10," "skin diseases associated with ultraviolet light," and "diseases associated with neuronal toxicity."

[0068] [Cosmetic Composition] The outer membrane vesicles derived from the specific lactic acid bacteria of the present invention can be used as an active ingredient in a cosmetic composition. Accordingly, one aspect of the present invention is a cosmetic composition containing outer membrane vesicles derived from lactic acid bacteria as an active ingredient. The cosmetic composition of the present invention is a cosmetic composition in which outer membrane vesicles derived from the specific lactic acid bacteria of the present invention activate FPR2, thereby exerting certain functions in the body (e.g., anti-inflammatory effects, epithelial cell regeneration and repair effects, etc.), i.e., a cosmetic composition for activating FPR2.

[0069] In addition to outer membrane vesicles derived from lactic acid bacteria, the cosmetic composition may optionally contain additives that are permitted to be incorporated into cosmetic compositions. Examples of additives include adjuvants and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances. Product forms of the cosmetic composition include cosmetics such as lotions, various creams, essences, and foundations, as well as facial cleansers, soaps, shampoos, treatments, and serums. Examples of dosage forms of the cosmetic composition include lotions, creams, essences, soaps, shampoos, treatments, foams, and foundations.

[0070] The content of outer membrane vesicles in the cosmetic composition can be appropriately determined taking into consideration the form of the cosmetic composition and the desired function. The content (number) of outer membrane vesicles per 1 mL of the cosmetic composition is, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12 The above explanation regarding the cosmetic composition also applies when the IL-10 inducer, skin protectant, and neuroprotectant according to the present invention are used as a cosmetic composition.

[0071] [Food and drink] The outer membrane vesicles derived from specific lactic acid bacteria of the present invention can be used as food and drink (food and drink). Accordingly, one aspect of the present invention is a food and drink containing outer membrane vesicles derived from lactic acid bacteria. Food and drink includes all foods and drinks that can be blended with outer membrane vesicles derived from lactic acid bacteria.

[0072] Specific examples of foods and beverages include functional foods and beverages (including those in various forms such as candy tablets, tablets, chewable tablets, tablets, powders, powders, capsules, granules, and drinks); beverages (tea drinks (green tea, oolong tea, black tea, etc.), soft drinks, jelly drinks, sports drinks, milk drinks, carbonated drinks, vegetable drinks, fruit juice drinks, fermented vegetable drinks, fermented fruit juice drinks, fermented milk drinks (yogurt, etc.), lactic acid bacteria drinks, milk drinks (coffee milk, fruit milk, etc.), powdered drinks, cocoa drinks, milk, and purified water); spreads (butter, jam, furikake, margarine, etc.); mayonnaise; shortening; custard cream; dressings; bread; cooked rice; noodles; pasta; miso soup; tofu; yogurt; soups or sauces; sweets (biscuits, cookies, chocolate, candy, cake, ice cream, chewing gum, tablets, etc.); and liquid diets (enteral nutritional supplements through tube feeding). Among these, functional foods and beverages are preferred. Functional foods and beverages refer to foods and beverages in which outer membrane vesicles derived from specific lactic acid bacteria of the present invention activate FPR2, thereby exerting certain functions in the body (e.g., anti-inflammatory effects, epithelial cell regeneration and repair effects, brain function improvement effects, etc.), i.e., FPR2-activating foods and beverages. Functional foods and beverages encompass so-called health foods and beverages in general. Examples of functional foods and beverages include health-claimed foods (foods for specified health uses (including conditional foods for specified health uses), functionally labeled foods, and foods with nutrient functions), special-use foods and beverages, dietary supplements, health supplements, supplements (including various dosage forms such as tablets, coated tablets, sugar-coated tablets, capsules, and liquids), and beauty foods and beverages (diet foods, etc.). Functional foods and beverages include health foods and beverages to which a health claim based on the Codex Alimentarius (Joint FAO / WHO Food Standards Commission) applies.

[0073] In addition to the lactic acid bacteria-derived outer membrane vesicles, the food and drink may contain any additives permitted for incorporation into food and drink. Examples of additives include colorants (sodium nitrite, etc.), coloring agents (gardenia pigment, Red 102, etc.), flavorings (orange flavoring, etc.), sweeteners (stevia, aspartame, etc.), preservatives (sodium acetate, sorbic acid, etc.), emulsifiers (sodium chondroitin sulfate, propylene glycol fatty acid ester, etc.), antioxidants (disodium EDTA, vitamin C, etc.), pH adjusters (citric acid, etc.), chemical seasonings (sodium inosinate, etc.), thickeners (xanthan gum, etc.), leavening agents (calcium carbonate, etc.), antifoaming agents (calcium phosphate), binders (sodium polyphosphate, etc.), nutritional fortifiers (calcium fortifier, vitamin A, etc.), and excipients (water-soluble dextrin, etc.).

[0074] Foods and beverages containing outer membrane vesicles derived from lactic acid bacteria can be produced by means available in the food industry. For example, outer membrane vesicles prepared in a liquid, gel, solid, powder, or granular form may be incorporated into the food or beverage. Alternatively, the outer membrane vesicles may be directly mixed, dissolved, or suspended in the raw materials of the food or beverage. The outer membrane vesicles may be applied, coated, impregnated, or sprayed onto the food or beverage. The outer membrane vesicles may be uniformly dispersed or unevenly distributed throughout the food or beverage. In the case of functional foods and beverages, the outer membrane vesicles may be encapsulated in a capsule, edible film, edible coating agent, or the like. In addition, in the case of functional foods and beverages, the outer membrane vesicles may be added with appropriate excipients and then molded into a shape such as a tablet. Foods and beverages containing outer membrane vesicles may be further processed, and such processed products are also within the scope of the present invention.

[0075] The content of outer membrane vesicles in foods and beverages can be appropriately determined taking into consideration the form of the food and beverage, the desired taste, etc. In the case of beverages, the content (number) of outer membrane vesicles per 1 mL of beverage is, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12The intake interval can be appropriately set depending on the type of food or beverage, and may be once a day or in divided doses. The above explanation regarding food or beverage also applies when the IL-10 inducer, skin protective agent, and neuroprotective agent according to the present invention are used as food or beverage.

[0076] [Feed] The outer membrane vesicles derived from specific lactic acid bacteria of the present invention can also be used as animal feed. Examples of feed include feed for livestock (such as cows and pigs), racehorses, and pets (such as dogs and cats). Since feed is almost the same as food and beverages except that the subjects are other than humans, the above description of food and beverages also applies to feed. The above description of feed also applies when the IL-10 inducer, skin protective agent, and neuroprotective agent according to the present invention are used as feed.

[0077] [Consumer Product Compositions] The outer membrane vesicles derived from specific lactic acid bacteria of the present invention can be used as an active ingredient in consumer product compositions. Accordingly, one aspect of the present invention is a consumer product composition containing outer membrane vesicles derived from lactic acid bacteria as an active ingredient. The consumer product composition of the present invention is a consumer product composition in which outer membrane vesicles derived from specific lactic acid bacteria of the present invention activate FPR2, thereby exerting certain functions in the body (e.g., anti-inflammatory effects, epithelial cell regeneration and repair effects, etc.), i.e., a consumer product composition for activating FPR2. Examples of consumer product compositions include deodorants, disinfectants, antiperspirants, fine fragrances such as perfumes, compositions for treating hard surfaces such as fabrics and flooring, and wipes, with deodorants and disinfectants being preferred. Examples of dosage forms of consumer product compositions include liquids, solids, semi-solids, gels, sprays, foams, liquid films, and foams.

[0078] In addition to the outer membrane vesicles derived from lactic acid bacteria, the consumer product composition may optionally contain additives that are permitted for incorporation into consumer product compositions, such as surfactants, chelating agents, dispersants, enzymes, enzyme stabilizers, brighteners, foam suppressors, dyes, fragrances, pigments, fabric softeners, and anti-agglomerating agents.

[0079] The content of OMV in the consumer product composition can be appropriately determined taking into consideration the dosage form. The content (number) of OMV per mL of the consumer product composition is, for example, 2 x 10 3 ~2x10 14 / mL, preferably 2x10 4 ~2x10 12 The above explanation regarding the consumer product composition also applies when the IL-10 inducer, skin protectant and neuroprotectant according to the present invention are used as a consumer product composition.

[0080] Next, the effects of the present invention will be specifically explained using examples, but the present invention is not limited to these examples.

[0081] [Test Bacterial Strains] The 26 types of bacteria shown in Table 1 were used.

[0082]

[0083] The bacteria numbers 1 to 14 and 20 to 26 were Lactobacillus. The bacteria numbers 15 to 16 were Lactococcus. The bacteria numbers 17 to 19 were Escherichia coli. The bacteria numbers 12 to 14 and 17 to 19 were used as comparative examples.

[0084] [Preparation Example: Preparation of Outer Membrane Vesicles Derived from Lactic Acid Bacteria] [Preparation of Outer Membrane Vesicles Derived from Lactobacillus] (1) Lactobacillus was pre-cultured (static culture at 37°C for 16 hours) using MRS liquid medium (BD Japan). The pre-culture solution was inoculated into MRS liquid medium (inoculation amount: 5% by volume) and subjected to main culture (static culture at 37°C for 16 hours). The main culture solution was centrifuged (Tomy Seiko, MX-301, 10,000g for 5 minutes).

[0085] (2) The supernatant after centrifugation was filtered through a polyvinylidene fluoride (PVDF) membrane filter (pore size: 0.45 μm). The filtrate was subjected to ultracentrifugation (Beckman Coulter, Optima XE-90, 150,000 g for 2 hours). The supernatant was removed after ultracentrifugation, and the resulting precipitate was suspended in 2.5 ml of phosphate-buffered saline (PBS) pre-filtered through a PVDF membrane filter (pore size: 0.1 μm) to obtain an OMV suspension. The OMV suspension was mixed with MRS liquid medium (BD Japan) pre-filtered through a PVDF membrane filter (pore size: 0.1 μm) at a volume ratio of 1:1. The mixture was treated with ExoQuick-TC® ULTRA (System Biosciences, LLC) and further filtered through a PVDF membrane filter (pore size: 0.45 μm) to obtain a purified OMV suspension.

[0086] (3) Based on the number of outer membrane vesicles determined according to the following quantification procedure, the purified outer membrane vesicle suspension was appropriately diluted with the buffer included in ExoQuick-TC (registered trademark) ULTRA (manufactured by System Biosciences, LLC) to obtain an outer membrane vesicle concentration of 6 x 10 8 A suspension containing 100 μl of OMV was prepared at a concentration of 1 / mL. This suspension was stored frozen (-20°C) until subjected to the activity test described below. [Quantification of OMV Number] The purified OMV suspension (100 μl) was diluted 10-fold with phosphate-buffered saline (PBS) containing 5.56 μM 3,3'-dioctadecyloxacarbocyanine perchlorate (a fluorescent probe for biomembrane labeling) and then incubated in the dark at 37°C for 2 hours to label the OMVs. The reaction solution was filtered through a PVDF membrane filter (pore size: 0.45 μm), and the filtrate was diluted 100-fold with PBS. The number of OMVs in the diluted solution was measured using the "Small particle detection mode" of an ultrasensitive, high-end benchtop flow cytometer (Cell Stream, Luminex). During the measurement, the laser output at wavelengths of 488 nm and 642 nm was set to 100%, and the output of forward and side scattered light was set to 0%. The number of particles detected at an excitation wavelength of 488 nm and a fluorescence wavelength of 528 nm was measured for 180 seconds, and the number of detected particles was taken as the number of outer membrane vesicles contained in the purified outer membrane vesicle suspension.

[0087] [Preparation of outer membrane vesicles derived from lactic acid bacteria] Lactococci were pre-cultured in GLM17 medium (static culture at 30°C for 16 hours). GLM17 medium was prepared by adding 0.5% glucose by mass and 0.5% lactose by mass to M17 liquid medium (manufactured by BD Japan). The pre-culture solution was inoculated into GLM17 medium (inoculation amount: 5% by volume) and subjected to main culture (static culture at 30°C for 16 hours). The main culture solution was centrifuged (Tomy Seiko MX-301, 10,000 g for 5 minutes). The supernatant after centrifugation was subjected to the procedures described in (2) and (3) of [Preparation of outer membrane vesicles derived from lactobacillus] above, until the outer membrane vesicle concentration reached 6 x 10 8 This suspension was stored frozen (-20°C) until subjected to activity testing.

[0088] [Preparation of outer membrane vesicles derived from Escherichia coli] Escherichia coli was pre-cultured using LB medium (Kanto Chemical) (shaking culture at 37°C for 16 hours). The pre-culture solution was inoculated into LB medium (inoculation amount: 5% by volume) and subjected to main culture (shaking culture at 37°C for 16 hours). The main culture solution was centrifuged (Tomy Seiko MX-301, 8,000g for 5 minutes). The supernatant after centrifugation was subjected to the procedures described in (2) and (3) of [Preparation of outer membrane vesicles derived from lactobacillus] above, until the outer membrane vesicle concentration reached 6 x 10 8 This suspension was stored frozen (-20°C) until subjected to activity testing.

[0089] Example 1: FPR2 agonist activity The FPR2 agonist activity of outer membrane vesicles was evaluated in a GPCR test commissioned by Tanso Biosciences (Cosmo Bio Co., Ltd.). In this test, a reporter assay was performed using the measurement principle described in International Publication No. WO2020 / 026979. As an evaluation sample, an outer membrane vesicle suspension (final concentration of outer membrane vesicles: 2 × 10) prepared from a frozen suspension (described above) was used. 6A blank sample was prepared according to the following procedure. MRS liquid medium (BD Japan) filtered through a PVDF membrane filter (pore size: 0.1 μm) was mixed with phosphate-buffered saline (PBS) filtered through a PVDF membrane filter (pore size: 0.1 μm) at a volume ratio of 1:1. The mixture was treated with ExoQuick-TC® ULTRA (System Biosciences, LLC) to obtain a blank sample. For the positive control sample, the peptide WKYMVM (Trp-Lys-Tyr-Met-Val-Met-NH2) (SEQ ID NO: 1) was used instead of outer membrane vesicles. This peptide is known to have FPR2 agonist activity (Molecules 2017, 22, 455). A sample containing only the solvent used in the test served as a negative control. The test was performed three times (n = 3 samples). The FPR2 agonist rate was calculated according to the following formula: The result of the operating rate is shown as the average of values ​​calculated from three tests.

[0090] FPR2 activation rate (%) = [(reporter luminescence amount by test sample - reporter luminescence amount by negative control sample) / (reporter luminescence amount by positive control sample - reporter luminescence amount by negative control sample)] x 100

[0091] Statistical analysis was performed using Student's t-test. The activity was compared with that of outer membrane vesicles derived from Escherichia coli T (bacterium number 17 in Table 1). A p<0.05 level was considered significant. The results are shown in Table 2 and Figure 1.

[0092] Table 2

[0093] As shown in Table 2 and Figure 1, outer membrane vesicles derived from lactic acid bacteria according to the present invention (bacterial numbers 1 to 11, 15 and 16, and 20 to 26) had FPR2 agonist activity. On the other hand, outer membrane vesicles derived from lactic acid bacteria other than the lactic acid bacteria according to the present invention and outer membrane vesicles derived from Escherichia coli did not have FPR2 agonist activity. Therefore, it was demonstrated that, among bacterial-derived outer membrane vesicles, outer membrane vesicles derived from specific types of lactic acid bacteria according to the present invention are useful as FPR2 agonists.

[0094] Example 2: FPR1 agonist activity The FPR1 agonist activity of outer membrane vesicles was evaluated in a GPCR test commissioned by Tanso Biosciences (Cosmo Bio Co., Ltd.). In this test, a reporter assay was performed using the measurement principle described in International Publication No. WO2020 / 026979. As an evaluation sample, an outer membrane vesicle suspension (final concentration of outer membrane vesicles: 2 × 10) prepared from a frozen suspension (described above) was used. 6 A blank sample was prepared according to the following procedure. MRS liquid medium (BD Japan) filtered through a PVDF membrane filter (pore size: 0.1 μm) was mixed with phosphate-buffered saline (PBS) filtered through a PVDF membrane filter (pore size: 0.1 μm) at a volume ratio of 1:1. The mixture was treated with ExoQuick-TC® ULTRA (System Biosciences, LLC) to obtain a blank sample. For the positive control sample, N-formylmethionyl-leucyl-phenylalanine (fMLP) was used instead of outer membrane vesicles. This peptide is known to have FPR1 agonist activity (Molecules 2017, 22, 455). A sample containing only the solvent used in the test served as a negative control. The test was performed three times (n = 3 data points). The FPR1 agonism rate was calculated according to the following formula. The agonism rate results are shown as the average of the values ​​calculated from the three tests. The results are shown in Table 3 and FIG.

[0095] FPR1 activating rate (%) = [(reporter luminescence amount by test sample - reporter luminescence amount by negative control sample) / (reporter luminescence amount by positive control sample - reporter luminescence amount by negative control sample)] x 100

[0096] Table 3

[0097] As shown in Table 3 and Figure 2, outer membrane vesicles derived from the lactic acid bacteria of the present invention (bacterial numbers 1 to 11, 15 and 16, and 20 to 26) did not have FPR1 agonist activity. Therefore, the outer membrane vesicles selectively agonized FPR2, demonstrating their usefulness as selective FPR2 agonists.

[0098] Reference Example 1: Effect of known FPR2 agonists on IL-10 production by M2 macrophages. The ability of M2 macrophages differentiated from human monocyte-derived cells (THP-1, JCRB Cell Bank) to induce the anti-inflammatory cytokine IL-10 was evaluated by the following procedure, which was a modification of a method described in a literature article (BMC Cancer 15:577 (2015)). THP-1 cells that had been passaged in a growth medium (RPMI 1640 medium (Nacalai Tesque, Inc.) supplemented with 10% by volume of fetal bovine serum (hiFBS, Cytiva) and 1% by volume of penicillin-streptomycin solution (Fujifilm Wako Pure Chemical Industries, Ltd.) that had been inactivated at 56°C for 30 minutes were cultured at 1 x 10 cells / mL in a growth medium supplemented with 150 nM phorbol 12-myristate 13-acetate (PMA, Sigma-Aldrich). 6The cells were suspended at a concentration of 1000 cells / ml and seeded in 0.1 ml aliquots onto a 96-well plate. The cells were cultured at 37°C under 5% CO2 for 24 hours. After incubation, the medium was removed, the plate was washed twice with growth medium, and 0.1 ml of growth medium was added. The cells were then cultured at 37°C under 5% CO2 for 24 hours. After incubation, the medium was removed, and 0.1 ml of growth medium supplemented with 20 ng / ml human IL-4 (R&D Systems) and 20 ng / ml human IL-13 (Thermo Fisher Scientific) was added. The cells were then cultured at 37°C under 5% CO2 for 24 hours. After incubation, the plate was washed twice with growth medium, and 0.1 ml of growth medium was added. A 2% test substance (described below) was added, and the plate was then cultured at 37°C under 5% CO2 for 24 hours. After incubation, the culture supernatant was collected, and IL-10 concentrations in the supernatant were calculated using the Human IL-10 DuoSet ELISA (R&D Systems). It is known that the IL-10-inducing ability of M2 macrophages is induced by Toll-like receptor 2 (TLR2) agonists (Arthritis Research & Therapy 19:245 (2017)). Therefore, three test samples were used: 500 ng / ml of the TLR2 agonist FSL-1 (InvivoGen) (FSL-1 group); a mixture of 500 ng / ml of FSL-1 and 5000 nM of the FLR2 agonist WKYMVm (Trp-Lys-Tyr-Met-Val-D-Met-NH2 (SEQ ID NO: 2), Abcam) (FSL-1 + WKYMVm group); and vehicle alone (negative control group). The test samples were added to the cells under test conditions at 2% by volume. Therefore, the final concentration of the test sample at the time of testing was 1 / 50 of that at the time of preparation. Statistical analysis was performed using Student's t-test. Tests were performed by comparing the IL-10 concentration with that of the FSL-1 group. A significant difference was considered to exist at a level of p<0.05. The number of data points per sample (n) was 6. The IL-10 concentration results are shown as the average of the values ​​measured in six tests. The results are shown in Table 4 and Figure 3.

[0099]

[0100] As shown in Table 4 and Figure 3, co-treatment of FSL-1 with an FPR2 agonist induced significantly higher levels of secretion of the anti-inflammatory cytokine IL-10 compared with treatment with the TLR2 agonist FSL-1 alone, demonstrating the importance of FPR2 agonism in the anti-inflammatory action of immune cells.

[0101] Example 3: IL-10 inducing activity. 8 A purified outer membrane vesicle suspension (test sample) derived from lactic acid bacteria was prepared according to the procedure described in the "Preparation Example" above, except that the concentration was adjusted to 1 / mL. The IL-10 induction activity of the test sample was evaluated according to the procedure described in the "Reference Example 1" above. A blank sample was prepared using the method described in the "Example 1" above. 500 ng / mL of FSL-1 (InvivoGen) was used as a positive control sample. A sample containing only the solvent used in the test was used as a negative control sample. The test sample was added to the cells under test at 2% by volume. Therefore, the final concentration of outer membrane vesicles during the test was 1 / 50 of that during preparation. The IL-10 induction rate was calculated according to the following formula.

[0102] IL-10 induction rate (%) = [(IL-10 concentration by addition of test sample - IL-10 concentration by addition of negative control sample) / (IL-10 concentration by addition of positive control sample - IL-10 concentration by addition of negative control sample)] x 100

[0103] Statistical analysis was performed using Student's t-test. The results were compared with the induction rate of outer membrane vesicles derived from Lactobacillus reuteri (bacterium number 12 in Table 1). A p<0.05 level was considered significant. The number of data points per sample (n) was 6. The IL-10 induction rate results are shown as the average of values ​​calculated from six tests. The results are shown in Table 5 and Figure 4.

[0104] Table 5

[0105] As shown in Table 5 and FIG. 4 , outer membrane vesicles derived from lactic acid bacteria with FPR2 agonist activity (bacterial numbers 1 to 4, 20, 21, 24, 25, 7, 9, 10, 26, 11, and 15 (Example 1)) showed significantly higher IL-10 induction rates than outer membrane vesicles derived from lactic acid bacteria without FPR2 agonist activity (bacterial number 12). Therefore, among bacterial-derived outer membrane vesicles, outer membrane vesicles derived from specific types of lactic acid bacteria according to the present invention were shown to be useful as IL-10 inducers via FPR2 agonism. This suggests that FPR2 agonism is important for the anti-inflammatory effect of outer membrane vesicles derived from specific types of lactic acid bacteria according to the present invention.

[0106] Example 4: Effect of FPR2 antagonist on IL-10 induction ability of outer membrane vesicles with FPR2 agonist activity. 9 Purified outer membrane vesicle (OVM) suspensions from Bacillus paracasei strain CP3526 (bacterium no. 9) were prepared according to the procedure described in the "Preparation Example" section above, except that the concentration was adjusted to 1 / mL. Three test samples were used: OVMs alone (CP3526 group), a mixture of OVMs and 50,000 nM of the FPR2 antagonist WRW4 (H-Trp-Arg-Trp-Trp-Trp-Trp-NH2 (SEQ ID NO: 3), Bachem AG) (CP3526 + WRW4 group), and solvent alone (negative control group). The test samples were added to the test cells at 2% by volume. Therefore, the final OVM concentration during the test was 1 / 50 of that at the time of preparation. IL-10 concentrations were evaluated according to the procedure described in "Reference Example 1" above. Statistical analysis was performed using Student's t-test. IL-10 concentrations were compared with those in the CP3526 group. A significant difference was considered at a p<0.05 level. The number of data points per sample (n) was 6. The IL-10 concentration results are shown as the average of values ​​measured in six tests. The results are shown in Table 6 and Figure 5.

[0107] Table 6

[0108] As shown in Table 6 and Figure 5, the IL-10 concentration was significantly lower in the CP3526+WRW4 group when an FPR2 antagonist was added compared to the CP3526 group when no FPR2 antagonist was added. This indicates that outer membrane vesicles derived from the Lacticaceae Bacillus paracasei CP3526 strain of the present invention induce IL-10 via FPR2 activation. This suggests that FPR2 agonism is required for the anti-inflammatory effect of outer membrane vesicles derived from specific types of lactic acid bacteria of the present invention in immune cells.

[0109] Example 5: Effect of protecting skin from ultraviolet rays. 10 Purified outer membrane vesicle suspensions derived from Bacillus paracasei strain CP3526 (bacterium No. 9) were prepared according to the procedure described in the "Preparation Example" above, except that the concentration was adjusted to 1.0 x 10 cells / mL. Three test samples were used: outer membrane vesicles alone (CP3526 group), a mixture of outer membrane vesicles and 50,000 nM of WRW4 (Bachem AG), an FPR2 antagonist (CP3526 + WRW4 group), and solvent alone (negative control group). Normal human adult epidermal keratinocytes (NHEK, KURABO) grown in Humedia-KG2 (KURABO) culture medium were cultured at a concentration of 1.0 x 10 cells / mL. 5 The cells were suspended at a concentration of 1000 cells / ml, and 0.1 ml of the suspension was seeded onto a 96-well black plate and cultured at 37°C for 24 hours in the presence of 5% CO2 by volume. After the culture, the plate was washed with PBS, and 0.02 ml of PBS was added. The plate was then irradiated with 75 mJ / cm2. 2The cells were irradiated with 10 ... The number of data points for each sample (n) was 3. The cell viability results are shown as the average of values ​​calculated from three tests. The results are shown in Table 7 and Figure 6.

[0110] Table 7

[0111] As shown in Table 7 and Figure 6, the addition of outer membrane vesicles significantly improved the survival rate of skin cells after UV irradiation compared to the absence of addition. Furthermore, the addition of an FPR2 antagonist significantly decreased the survival rate compared to the absence of addition. Therefore, it was demonstrated that outer membrane vesicles derived from the Lacticaceae Bacillus paracasei CP3526 strain according to the present invention protect skin from UV rays through the activation of FPR2. This suggests that the protective effect of outer membrane vesicles derived from specific types of lactic acid bacteria according to the present invention on skin cells requires FPR2 agonist activity.

[0112] Example 6: Effect of protecting neurons from neurotoxicity. 10A purified outer membrane vesicle suspension from Bacillus paracasei strain CP3526 (bacterium No. 9) was prepared according to the procedure described in the "Preparation Example" section above, except that the concentration was adjusted to cells / mL. Two test samples were used: outer membrane vesicles alone (CP3526 group) and solvent alone (negative control group). PC-12 cells (ECACC) grown in a growth induction medium (growth induction medium) consisting of DMEM (1.0 g / L glucose) (containing L-glutamine and pyruvate) (liquid) (DMEM (LG), Nacalai Tesque) supplemented with 5% by volume of fetal bovine serum (FBS, Thermo Fisher Scientific), 10% by volume of horse serum (hiHS, Thermo Fisher Scientific) inactivated at 56°C for 30 minutes, and 1% by volume of penicillin-streptomycin solution (P / S) (Nacalai Tesque) were added. At a concentration of 8 x 10 cells / mL, the cells were cultured in a 2000 ml / ml medium containing 10% by volume of penicillin-streptomycin solution (P / S) (Nacalai Tesque). 4The cells were suspended at a concentration of 1000 cells / ml and seeded in 0.1 ml onto a Collagen IV-coated 96-well plate (Corning) and cultured at 37°C under 5% CO2. On the day after and on the fourth day after seeding, the medium was replaced with differentiation medium (DMEM (LG) supplemented with 0.1% FBS, 0.1% hiHS, 1% P / S, and 50 ng / ml mouse NGF 2.5S native protein (NGF, Thermo Fisher Scientific)) to induce neuronal differentiation. On the fifth day after seeding, the medium was replaced with differentiation medium containing 2% test sample and cultured at 37°C under 5% CO2 for 1 hour. The medium was then replaced with differentiation medium containing 60 μM β Amyloid 1-42 (Sigma-Aldrich) (neuronal cytotoxicity) and 2% test sample, and cultured at 37°C under 5% CO2 for 24 hours. After incubation, intracellular ATP levels were measured. The measurement method was as follows: 0.1 ml / well of "Cell" ATP Assay Reagent Ver. 2 (Toyo B-Net Co., Ltd.) was added and shaken for 1 minute using a plate shaker. After 10 minutes of incubation at 23°C, 100 μl of the solution was transferred to a 96-well black plate and the luminescence intensity was measured. Each luminescence intensity was divided by the average luminescence intensity measured in five replicates without amyloid beta treatment, and the result was multiplied by 100 to determine cell viability. Statistical analysis was performed using Student's t-test. Cell viability was compared with that of the CP3526 group. A p<0.05 level was considered significant. The number of data points per sample (n) was 5. Cell viability results are shown as the average of the values ​​calculated for five replicates. The results are shown in Table 8 and Figure 7.

[0113] Table 8

[0114] As shown in Table 8 and Figure 7, the addition of outer membrane vesicles significantly improved the survival rate of neurons after amyloid-β treatment compared to the absence of addition. Therefore, it was demonstrated that outer membrane vesicles derived from the Lacticaceae Bacillus paracasei CP3526 strain according to the present invention protect neurons from neurotoxicity. This suggests that outer membrane vesicles derived from a specific type of lactic acid bacteria according to the present invention have a useful effect against neurotoxicity-induced damage in neurons.

[0115] The present invention can be used for medicines, foods and beverages, etc.

Claims

1. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus An FPR2 agonist containing outer membrane vesicles derived from at least one lactic acid bacterium selected from the group consisting of the following.

2. Lactic acid bacteria, as follows: Lactobacillus delbrueckii subsp. bulgaricus Lactobacillus helveticus Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei Lactiplantibacillus plantarum Ligilactobacillus salivarius Lactococcus lactis subsp. lactis Streptococcus salivarius subsp. thermophilus Lactobacillus gasseri Lactobacillus amylovorus Lactobacillus crispatus Lactobacillus johnsonii Lacticaseibacillus casei, and Lactiplantibacillus pentosus The FPR2 agonist according to claim 1, which is at least one selected from the group consisting of the following.

3. An FPR2 agonist according to claim 1 or 2, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonistic activity.

4. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A food or beverage for FPR2 activation containing, as an active ingredient, an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following:

5. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A pharmaceutical composition for treating diseases associated with FPR2, comprising, as an active ingredient, an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

6. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A cosmetic composition for FPR2 activation, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

7. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A consumer product composition for FPR2 activation, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

8. The food or beverage according to claim 4, the pharmaceutical composition according to claim 5, the cosmetic composition according to claim 6, or the consumer product composition according to claim 7, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 activating activity.

9. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A food or beverage for protecting the skin from ultraviolet rays, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

10. Lactobacillus helveticus, Lacticaseibacillus paracasei Ligilactobacillus salivarius Lactobacillus gasseri Lactobacillus amylovorus Lactobacillus crispatus Lactobacillus johnsonii Lacticaseibacillus casei, and Lactiplantibacillus pentosus A pharmaceutical composition for protecting the skin from ultraviolet rays, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

11. Lactobacillus helveticus, Lacticaseibacillus paracasei Ligilactobacillus salivarius Lactobacillus gasseri Lactobacillus amylovorus Lactobacillus crispatus Lactobacillus johnsonii Lacticaseibacillus casei, and Lactiplantibacillus pentosus A cosmetic composition for protecting the skin from ultraviolet rays, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

12. Lactobacillus helveticus, Lacticaseibacillus paracasei Ligilactobacillus salivarius Lactobacillus gasseri Lactobacillus amylovorus Lactobacillus crispatus Lactobacillus johnsonii Lacticaseibacillus casei, and Lactiplantibacillus pentosus A consumer product composition for protecting the skin from ultraviolet rays, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

13. Food or beverage according to claim 9, pharmaceutical composition according to claim 10, cosmetic composition according to claim 11, or consumer product composition according to claim 12, which protect the skin from ultraviolet rays through FPR2 operation.

14. The food or beverage according to claim 9, the pharmaceutical composition according to claim 10, the cosmetic composition according to claim 11, or the consumer product composition according to claim 12, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 activating activity.

15. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus Lactobacillus acidophilus, Lactobacillus delbrueckii Lactobacillus crispatus Lactobacillus gasseri Lactobacillus paragasseri Lactobacillus amylovorus Lactobacillus johnsonii Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A food or beverage for protecting nerve cells from neurotoxicity, containing, as an active ingredient, an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following:

16. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus Lactobacillus acidophilus, Lactobacillus delbrueckii Lactobacillus crispatus Lactobacillus gasseri Lactobacillus paragasseri Lactobacillus amylovorus Lactobacillus johnsonii Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A pharmaceutical composition for protecting nerve cells from neurotoxicity, containing, as an active ingredient, an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

17. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus Lactobacillus acidophilus, Lactobacillus delbrueckii Lactobacillus crispatus Lactobacillus gasseri Lactobacillus paragasseri Lactobacillus amylovorus Lactobacillus johnsonii Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A cosmetic composition for protecting nerve cells from neurotoxicity, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

18. Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus helveticus Lactobacillus acidophilus, Lactobacillus delbrueckii Lactobacillus crispatus Lactobacillus gasseri Lactobacillus paragasseri Lactobacillus amylovorus Lactobacillus johnsonii Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A consumer product composition for protecting nerve cells from neurotoxicity, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

19. Food or beverage according to claim 15, pharmaceutical composition according to claim 16, cosmetic composition according to claim 17, or consumer product composition according to claim 18, which protect nerve cells from neurotoxicity by FPR2 activation.

20. The food or beverage according to claim 15, the pharmaceutical composition according to claim 16, the cosmetic composition according to claim 17, or the consumer product composition according to claim 18, wherein the neurotoxicity is due to amyloid-beta.

21. Food and beverage according to claim 15, pharmaceutical composition according to claim 16, cosmetic composition according to claim 17, or consumer product composition according to claim 18, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 activating activity.

22. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A food or beverage for inducing IL-10 by FPR2 activation, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

23. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A pharmaceutical composition for treating diseases associated with IL-10 by FPR2 activation, comprising, as an active ingredient, an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following:

24. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A cosmetic composition for inducing IL-10 by FPR2 activation, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

25. Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A consumer product composition for IL-10 induction by FPR2 activation, containing as an active ingredient an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

26. The food or beverage according to claim 22, the pharmaceutical composition according to claim 23, the cosmetic composition according to claim 24, or the consumer product composition according to claim 25, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 activating activity.

27. Lacticaseibacillus paracasei strain CP3526, with accession number NITE BP-03625.

28. A method for activating FPR2 in the subject, Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A method (excluding human treatment methods) comprising the step of administering to a subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

29. A method for treating diseases associated with FPR2 in the subject, Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A method (excluding human treatment methods) comprising the step of administering to a subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

30. A method for protecting the skin from ultraviolet rays, Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A method (excluding human treatment methods) comprising the step of administering to a subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

31. The method according to claim 30, wherein the FPR2 function protects the skin from ultraviolet rays.

32. A method for protecting nerve cells from neurotoxicity in the subject, Lactobacillus helveticus Lacticaseibacillus paracasei Ligilactobacillus salivarius Lactobacillus gasseri Lactobacillus amylovorus Lactobacillus crispatus Lactobacillus johnsonii Lacticaseibacillus casei, and Lactiplantibacillus pentosus A method (excluding human treatment methods) comprising the step of administering to a subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

33. The method according to claim 32, wherein FPR2 activation protects nerve cells from neurotoxicity.

34. A method for inducing IL-10 by FPR2 activation in the target, Lactobacillus genus, Lacticaseibacillus genus, Lactiplantibacillus genus, Ligilactobacillus genus, Lactococcus genus, and Streptococcus salivarius subsp. thermophilus A method (excluding human treatment methods) comprising the step of administering to a subject an outer membrane vesicle derived from at least one lactic acid bacterium selected from the group consisting of the following.

35. Lactic acid bacteria, as follows: Lactobacillus delbrueckii subsp. bulgaricus Lactobacillus helveticus Lactobacillus acidophilus, Lacticaseibacillus rhamnosus, Lacticaseibacillus paracasei Lactiplantibacillus plantarum Ligilactobacillus salivarius Lactococcus lactis subsp. lactis Streptococcus salivarius subsp. thermophilus Lactobacillus gasseri Lactobacillus amylovorus Lactobacillus crispatus Lactobacillus johnsonii Lacticaseibacillus casei, and Lactiplantibacillus pentosus The method according to any one of claims 28 to 31 and 34, wherein at least one is selected from the group consisting of the following.

36. The method according to any one of claims 28 to 31, wherein the outer membrane vesicles derived from lactic acid bacteria do not have FPR1 agonizing activity.