Agent for preventing or improving skin bacterial infections

Abstract

To provide a material useful for suppressing skin infection caused by Staphylococcus aureus and preventing or improving skin bacterial infection.SOLUTION: Provided is a Staphylococcus aureus infection inhibitor comprising, as an active ingredient, bacterial cells of the genus Neisseria or a culture of the bacterial cells.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to an agent for preventing or improving skin bacterial infections.

Background Art

[0002] The skin serves as a barrier against bacterial infections and normally does not develop bacterial infections. However, in addition to skin injury sites caused by burns, cracks, surgical wounds, catheter insertion, etc., in patients with atopic dermatitis, diabetes, or immune diseases, bacterial infections often occur, causing severe inflammation in the epidermis, dermis, or subcutaneous layer of the skin. The causative bacteria of such skin infections are mainly known as Staphylococcus and Streptococcus, and in particular, infections caused by Staphylococcus aureus (hereinafter referred to as Staphylococcus aureus) are common.

[0003] For example, it has been reported that the formation of Staphylococcus aureus colonies increases in the skin of patients with atopic dermatitis compared to healthy individuals (Non-Patent Documents 1 and 2), and it has been reported that there is a correlation between the average density of S. aureus in the skin and the severity (Skin grade of AD patients) (Non-Patent Document 2).

[0004] In addition, it has also been reported that skin infections caused by Staphylococcus aureus cause damage to the skin barrier and promote inflammation (Non-Patent Document 3). Furthermore, it has been reported that infections caused by Staphylococcus aureus promote the activity of endogenous proteases kallikreins (KLKs), promote the proteolysis of desmoglein-1 and filaggrin, which are proteins that constitute the skin barrier function, and cause damage to the skin barrier (Non-Patent Document 4). In patients with atopic dermatitis, it has also been reported that Staphylococcus aureus causes a decrease in skin barrier function, penetrates into the dermis through the epidermal barrier, and leads to exacerbation of atopy (Non-Patent Document 5).

[0005] Therefore, suppressing infections caused by Staphylococcus aureus is important for the prevention or treatment of skin bacterial infections. For the prevention or treatment of skin bacterial infections, bactericides and antibiotics against the causative bacteria can be mentioned. However, due to their strong bactericidal and bacteriostatic effects, these technologies may destroy the normal skin flora and cause the emergence of resistant bacteria, leading to serious diseases.

[0006] On the other hand, bacteria of the genus Neisseria, such as Neisseria mucosa, Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, and Neisseria elongata, are known to exist as non-pathogenic commensal bacteria in the oral cavity, but their functions are not clear.

Prior Art Documents

Non-Patent Documents

[0007]

Non-Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0008] The present invention relates to providing a material useful for suppressing skin infections caused by Staphylococcus aureus and for preventing or improving skin bacterial infections.

Means for Solving the Problems

[0009] The present inventors have found that cells or cultures of bacteria of the genus Neisseria, known as non-pathogenic oral commensal bacteria, suppress cell infections caused by Staphylococcus aureus and are useful as a prophylactic or therapeutic agent for skin bacterial infections.

[0010] That is, the present invention relates to the following 1) to 4). 1) An inhibitor for suppressing Staphylococcus aureus infection, comprising cells or cell cultures of bacteria of the genus Neisseria as an active ingredient. 2) A prophylactic or ameliorating agent for skin bacterial infections, comprising cells or cell cultures of bacteria of the genus Neisseria as an active ingredient. 3) A food for suppressing Staphylococcus aureus infection, comprising cells or cell cultures of bacteria of the genus Neisseria as an active ingredient. 4) A food for preventing or improving skin bacterial infections, comprising cells or cell cultures of bacteria of the genus Neisseria as an active ingredient.

Effects of the Invention

[0011] Since the Neisseria bacteria of the present invention are non-pathogenic oral commensal bacteria, they can provide an agent for preventing or improving skin bacterial infections that can be ingested by healthy people as well as the elderly and convalescents over a long period of time.

Brief Description of the Drawings

[0012]

Figure 1

Figure 2

Modes for Carrying Out the Invention

[0013] Examples of the Neisseria bacteria used in the present invention include Gram-negative cocci belonging to the genus Neisseria of the family Neisseria. Specifically, Neisseria mucosa, Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens, Neisseria elongata, etc. may be mentioned, and Neisseria mucosa is preferably used. All of these are known as non-pathogenic oral commensal bacteria and can be isolated from human saliva, dental plaque, etc. In addition, predetermined microbial strains can be obtained from public microbial depository institutions (Neisseria mucosa: JCM12992 strain, Neisseria sicca: ATCC29256 strain, Neisseria elongata: ATCC25295 strain, Neisseria flava: ATCC14221 strain, Neisseria subflava: ATCC49275 strain, Neisseria flavescens: ATCC13120 strain).

[0014] In the present invention, the "bacterial cells" of the genus Neisseria may be any of viable cells, moist cells, or dried cells of the genus Neisseria. Further, not only viable cells but also dead cells may be used. In addition, the bacterial cells include cell components such as cytoplasm and cell wall components obtained by treating the bacterial cells with enzymes or physical means. Examples of the "bacterial cell culture" of the genus Neisseria include cultures containing medium components, bacterial cells, and extracellular polymeric substances (EPS), etc., cultures containing bacterial cells and EPS from which the medium components have been removed, cultures containing medium components and EPS from which the bacterial cells have been removed, or cultures containing EPS from which the medium components and bacterial cells have been removed. The bacterial cells or bacterial cell culture of the present invention can be prepared in any form according to the purpose of use, such as freeze-dried powder, spray-dried powder, suspension in a liquid, etc., if desired.

[0015] The medium for culturing the Neisseria bacteria of the present invention is not particularly limited as long as it contains peptone or the like as a nutrient source, and examples thereof include BHI medium (manufactured by BD Japan). The culturing method is not particularly limited as long as it is a condition under which the bacterial cells of the genus Neisseria grow well. For example, culturing can be carried out at 20 to 40 °C, preferably 35 to 38 °C, under aerobic conditions, more preferably under a CO2 concentration of 5% or under microaerobic conditions for 16 to 72 hours.

[0016] In the present invention, when using the cell culture of bacteria of the genus Neisseria, for the culture of bacteria of the genus Neisseria, the culture supernatant of periodontal pathogenic bacteria (for example, Porphyromonas gingivalis, Fusobacterium nucleatum, Prevotella intermedia, Treponema denticola, Tannerella forsythia, Aggregatibacter actinomycetemcomitans, Campylobacter rectus, Selenomonas sputigena, etc.) is used. Preferably, Porphyromonas gingivalis is cultured under anaerobic conditions, and the culture supernatant obtained by removing the cells from the obtained culture solution by centrifugation or the like is added. The culture method after addition is not particularly limited as long as EPS is produced in a sufficient amount. From the viewpoint of EPS production, 20 to 40 °C is preferable, and more preferably 35 to 38 °C. It is preferably carried out for 24 hours or more under aerobic conditions, more preferably under a CO2 concentration of 5% or under microaerobic conditions. Also, from the viewpoint of preventing the contamination of Porphyromonas gingivalis cells and cell debris in EPS, it is more preferable to add the culture supernatant after sterilization by filter sterilization or the like and carry out the culture at 35 to 38 °C under a CO2 concentration of 5% or under microaerobic conditions for 24 hours or more. The addition amount of the culture supernatant of periodontal pathogenic bacteria is preferably 0.05 v / v% or more, more preferably 0.5 v / v% or more, and further preferably 5 v / v% or more from the viewpoint of the EPS production induction effect. Also, from the viewpoint of the EPS recovery amount, it is preferably 50 v / v% or less, more preferably 30 v / v% or less, and further preferably 10 v / v% or less.

[0017] In the present invention, as the cell culture of bacteria of the genus Neisseria, a cell culture of Neisseria mucosa is preferably used, and it is more preferable that Neisseria mucosa is cultured in a culture solution obtained by culturing Porphyromonas gingivalis in an anaerobic environment.

[0018] As shown in the following examples, the cells or cell cultures of bacteria of the genus Neisseria have an inhibitory effect on the infection of Staphylococcus aureus to HeLa cells. Therefore, the cells or cell cultures of bacteria of the genus Neisseria can be used as an inhibitor for suppressing Staphylococcus aureus infection and an agent for preventing or improving skin bacterial infections, and the cells or cell cultures of bacteria of the genus Neisseria can be used for manufacturing an inhibitor for suppressing Staphylococcus aureus infection and an agent for preventing or improving skin bacterial infections. In addition, the cells or cell cultures of bacteria of the genus Neisseria can be used for suppressing Staphylococcus aureus infection and for preventing or improving skin bacterial infections. Here, such use can be administration to humans or non-human animals, or use in specimens derived from them, and can be either therapeutic use or non-therapeutic use. Here, examples of non-human animals include non-human mammals, amphibians, and cartilaginous fish, and examples of non-human mammals include, for example, anthropoid apes, other primates, mice, rats, horses, cows, pigs, sheep, dogs, cats, hamsters, and companion animals.

[0019] In the present invention, "skin bacterial infection" mainly means a skin infection caused by Staphylococcus aureus, and examples include cellulitis, erysipelas, impetigo contagiosa, folliculitis, deep skin infection, boils, carbuncles, and suppurative paronychia. In the present invention, Staphylococcus aureus means Staphylococcus aureus, which is a facultative anaerobic Gram-positive coccus, and includes drug-resistant Staphylococcus aureus (for example, methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Staphylococcus aureus (VRSA), etc.). In addition, "Staphylococcus aureus infection suppression" means suppressing the infection of Staphylococcus aureus into animal cells, and includes infection inhibition.

[0020] In this specification, "prevention" means preventing or delaying the onset of a disease or symptom in an individual, or reducing the risk of onset of a disease or symptom in an individual. In addition, "improvement" means the improvement of a disease, symptom or condition, the prevention or delay of the deterioration of a disease, symptom or condition, or the reversal, prevention or delay of the progression of a disease or symptom, and includes the so-called "treatment" concept.

[0021] The Staphylococcus aureus infection inhibitor, the preventive or improvement agent for skin bacterial infections of the present invention can itself be a pharmaceutical, quasi-drug, or food for suppressing Staphylococcus aureus infection and for preventing or improving skin bacterial infections, and can also be a material or preparation to be formulated and used in said pharmaceutical, quasi-drug, or food. Note that foods include foods conceptually for preventing or improving skin bacterial infections, and which display to that effect as necessary, functional display foods, foods for specified health use, foods for patients, and supplements.

[0022] When used as a pharmaceutical (including quasi-drugs), it may be in any form of oral administration or parenteral administration. Dosage forms for oral administration include, for example, liquid preparations; solid preparations such as tablets, granules, fine granules, powders, and tablets; or various forms such as capsules encapsulating the liquid preparation or solid preparation, oral sprays, and lozenges. Dosage forms for parenteral administration include various preparations such as topical skin, transdermal, transmucosal, patch, nasal, enteral, suppository, injection, and inhalation. Among these, a preferred dosage form is a topical skin preparation, specifically, forms such as ointments, emulsions, creams, milky lotions, lotions, gels, and aerosols. Such various dosage forms of pharmaceutical preparations can be prepared by appropriately combining other pharmaceutically acceptable excipients, binders, extenders, disintegrants, surfactants, lubricants, dispersants, buffers, preservatives, flavoring agents, fragrances, coating agents, carriers, diluents, etc. within the range that does not interfere with the action of the cells or cell cultures of *Neisseria mucosa*.

[0023] When used as a food, the forms include beverages such as fruit juice or vegetable juice beverages, carbonated beverages, tea-based beverages, milk beverages, fermented milk, fermented fruit juice, fermented vegetable juice, alcoholic beverages, and soft drinks, as well as jelly-like foods, various snacks, baked goods, cakes, chocolates, jams, breads, gums, candies, soups, pickles, simmered foods, etc. In addition, supplements in the same forms as the above-mentioned oral administration preparations (tablets, capsules, syrups, etc.) are included. The food can be prepared according to a conventional method by appropriately combining the cells or cell cultures of *Neisseria* bacteria with any food material, or other active ingredients, or additives acceptable in foods (for example, solvents, softeners, oils, emulsifiers, preservatives, acidulants, sweeteners, bittering agents, pH adjusters, stabilizers, colorants, ultraviolet absorbers, antioxidants, humectants, thickeners, fixing agents, dispersants, fluidity improvers, wetting agents, fragrances, seasonings, flavor modifiers, etc.).

[0024] The content of the cells of *Neisseria* bacteria in the above-mentioned pharmaceutical (including quasi-drugs) or food is not particularly limited, but it may be appropriately adjusted according to the daily dosage, etc.

[0025] In the above-mentioned pharmaceuticals (including quasi-drugs) or foods, the dosage of the cells of bacteria of the genus Neisseria can be appropriately determined according to various conditions such as the patient's body weight, age, gender, symptoms, etc. For example, when expressed in terms of the number of bacteria of the genus Neisseria, the daily dosage is preferably 5×10 8 or more, more preferably 1×10 10 or more, still more preferably 1×10 11 or more, and preferably 1×10 14 or less, more preferably 1×10 13 or less, still more preferably 1×10 12 or less. Also, the dosage of the cell culture of bacteria of the genus Neisseria is preferably 1 mg or more, more preferably 100 mg or more, still more preferably 500 mg or more, and preferably 5000 mg or less, more preferably 2500 mg or less, still more preferably 1000 mg or less per day for an adult in terms of dry weight.

[0026] The administration or ingestion targets of the Staphylococcus aureus infection inhibitor, the food for inhibiting Staphylococcus aureus infection, the prophylactic or ameliorating agent for skin bacterial infections, and the food for prophylactic or ameliorating skin bacterial infections of the present invention are preferably humans suffering from skin bacterial infections, humans desiring prevention of the onset of skin bacterial infections, for example, atopic dermatitis patients at risk of developing skin bacterial infections, humans desiring to suppress the decrease or vulnerability of skin barrier function due to skin bacterial infections, and the like.

[0027] Regarding the above-described embodiments, the following aspects are further disclosed in the present invention. <1> A Staphylococcus aureus infection inhibitor containing, as an active ingredient, the cells or cell culture of bacteria of the genus Neisseria. <2> A prophylactic or ameliorating agent for skin bacterial infections containing, as an active ingredient, the cells or cell culture of bacteria of the genus Neisseria. <3> A food for inhibiting Staphylococcus aureus infection containing, as an active ingredient, the cells or cell culture of bacteria of the genus Neisseria. <4> A food for prophylactic or ameliorating skin bacterial infections containing, as an active ingredient, the cells or cell culture of bacteria of the genus Neisseria.

[0028] <5>Use of a cell or cell culture of a bacterium of the genus Neisseria for producing an agent for suppressing Staphylococcus aureus infection. <6>Use of a cell or cell culture of a bacterium of the genus Neisseria for producing an agent for preventing or improving skin bacterial infections. <7>Use of a cell or cell culture of a bacterium of the genus Neisseria for producing a food for suppressing Staphylococcus aureus infection. <8>Use of a cell or cell culture of a bacterium of the genus Neisseria for producing a food for preventing or improving skin bacterial infections.

[0029] <9>A cell or cell culture of a bacterium of the genus Neisseria for use in suppressing Staphylococcus aureus infection. <10>A cell or cell culture of a bacterium of the genus Neisseria for use in preventing or improving skin bacterial infections.

[0030] <11>Non-therapeutic use of a cell or cell culture of a bacterium of the genus Neisseria for suppressing Staphylococcus aureus infection. <12>Non-therapeutic use of a cell or cell culture of a bacterium of the genus Neisseria for preventing or improving skin bacterial infections.

[0031] <13>A method for suppressing Staphylococcus aureus infection, which comprises administering or ingesting a cell or cell culture of a bacterium of the genus Neisseria in an effective amount to a subject in need thereof. <14>A method for preventing or improving skin bacterial infections, which comprises administering or ingesting a cell or cell culture of a bacterium of the genus Neisseria in an effective amount to a subject in need thereof.

[0032] <15>In <1> to <14>, the bacterium of the genus Neisseria is at least one selected from Neisseria mucosa, Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens and Neisseria elongata, preferably Neisseria mucosa. <16><2>, <4>, <6>, <8>, <10>, <12> and <14>, the skin bacterial infection is a skin infection caused by Staphylococcus aureus. <17><1>, <3>, <5>, <7>, <9>, <11>, <13> and <16>, Staphylococcus aureus is a drug-resistant Staphylococcus aureus. <18><1> to <14>, the bacterial culture is preferably a culture obtained by adding the culture supernatant of Porphyromonas gingivalis to the culture solution of Neisseria mucosa and culturing, or a culture containing EPS obtained by adding the culture supernatant of Porphyromonas gingivalis to the culture solution of Neisseria mucosa, culturing, and removing the bacterial cells and medium components.

Example

[0033] Example 1 Inhibitory effect of Neisseria bacteria on Staphylococcus aureus infection (1) Cell culture conditions HeLa cells were obtained from the American Type Culture Collection (ATCC). HeLa cells were cultured at a temperature of 37°C and a CO2 concentration of 5% using DMEM / F12 (Gibco, containing 10 v / v% FBS, 1 v / v% Penicillin-Streptomycin).

[0034] (2) Bacterial culture conditions Staphylococcus aureus was obtained from the National Institute of Technology and Evaluation, and six strains of Neisseria bacteria were obtained from ATCC or the RIKEN BioResource Center, National Institute of Advanced Industrial Science and Technology (Table 1). For all bacteria, colony culture was performed using Brain Heart Infusion (BHI) Agar medium (Nippon Becton Dickinson (hereinafter, Nippon BD)), and liquid culture was performed using BHI medium (Nippon BD). Staphylococcus aureus was aerobically cultured at a temperature of 37°C, and Neisseria bacteria were cultured under conditions of a temperature of 37°C and a CO2 concentration of 5%. After 24 hours of liquid culture, Staphylococcus aureus cells were further cultured in liquid at a 1000-fold dilution for another 24 hours. After 24 hours of liquid culture, bacteria of the genus Neisseria were further cultured in liquid at a 100-fold dilution for 48 hours, and the resulting cells were used.

[0035]

Table 1

[0036] (3) Reagents The labeling reagent Carboxyfluorescein diacetate succinimidyl ester (CFSE) was purchased from Dojindo Laboratories, dissolved in DMSO, and used at a final concentration of 10 μM.

[0037] (4) Method for fluorescent labeling of bacterial cells The culture solution of Staphylococcus aureus was centrifuged at 22 °C and 4,500 g for 10 minutes to collect the precipitated cells. The collected cells were washed with PBS(-) and then with DMEM / F12 (Gibco) by centrifugation at 22 °C and 4,500 g for 5 minutes, adjusted to OD600 = 2.0, and incubated in a 10 μM CFSE solution (dissolved in DMEM / F12) at 37 °C in the dark for 30 minutes for fluorescent labeling. The fluorescently labeled cells were washed twice with DMEM / F12 (Gibco) by centrifugation at 22 °C and 4,500 g for 5 minutes, and a solution of Staphylococcus aureus cells labeled with CFSE was prepared by adjusting the turbidity of the cell suspension.

[0038] (5) Infection method After removing the medium from confluent HeLa cells in a 12-well plate, the cells were washed with PBS(-). Various Neisseria bacteria and CFSE-labeled Staphylococcus aureus were suspended in DMEM / F12 (Gibco) and added at a multiplicity of infection (MOI) = 500, followed by co-culture for 2 hours to infect the cells. As a control group, a group without the addition of Neisseria bacteria (Control) was also prepared.

[0039] (6) Analytical method for infected cells The medium was removed from HeLa cells infected with CFSE-labeled Staphylococcus aureus, washed twice with PBS(-), and then the infected cells were detached using 400 μL of 0.25% Trypsin-EDTA (Gibco). 750 μL of DMEM / F12 (Gibco, containing 10 v / v% FBS) was added, the total volume of the solution was collected in a 1.5 mL tube, and the infected cells were collected by centrifugation at 4°C, 400 g for 5 minutes. The supernatant was discarded, 400 μL of 4% paraformaldehyde phosphate buffer (Wako) was added and pipetted, and fixed at 4°C for 10 minutes. After adding 800 μL of PBS (containing 2 v / v% FBS) and pipetting, centrifugation was performed at 4°C, 600 g for 5 minutes, and suspended in 200 μL of PBS (containing 2 v / v% FBS) to obtain an analysis sample. Flow cytometry (BD, FACSVerse) was used for the analysis. Based on forward scatter (FSC) and side scatter (SSC), the mean fluorescence intensity (MFI) was calculated from the fluorescence intensity obtained in the FITC channel of 20,000 gated cells, and the cell infectivity was evaluated by detecting the cells adhered and infected with fluorescently labeled Staphylococcus aureus. The results are shown in Figure 1.

[0040] (7) Statistical analysis The obtained results were shown as dots for the MFI of each sample, and the mean ± standard deviation (Mean ± standard deviation (SD)) of each group was represented by a bar graph. The test for the difference in means was performed using the statistical analysis software SPSS (IBM) with Dunnett's test. When the P value was less than 0.05 (*p < 0.05), less than 0.01 (**p < 0.01), or less than 0.001 (***p < 0.001), it was considered a statistically significant difference.

[0041] (8) Results In the evaluation of cell infectivity, a significant decrease in Mean fluorescence intensity (cell infectivity) was observed with Neisseria mucosa (N. mucosa), Neisseria sicca (N. sicca), Neisseria elongata (N. elongata), Neisseria flava (N. flava), Neisseria subflava (N. subflava), and Neisseria flavescens (N. flavescens) compared to the control (Figure 1).

[0042] Example 2 Inhibitory effect of Neisseria mucosa cell culture on Staphylococcus aureus infection (1) Preparation of Neisseria mucosa cell culture A culture solution of Neisseria mucosa was prepared by culturing Neisseria mucosa in 300 mL of BHI medium at 37°C and 5% CO2 for 48 hours. The culture supernatant of Porphyromonas gingivalis prepared to a final concentration of 5 v / v% was added to the Neisseria mucosa culture solution, and after culturing at 37°C and 5% CO2 for 48 hours, 20 mL of the culture solution was collected in a 50 mL tube. The culture supernatant of Porphyromonas gingivalis was obtained by culturing colonies grown by culturing Porphyromonas gingivalis (ATCC 33277 strain) obtained from ATCC on an anaerobic Columbia agar medium (BD, Japan) at 37°C under anaerobic conditions for 24 hours or more in GAM broth medium (Nissui Pharmaceutical, containing 5.0 μg / mL hemin, 17.4 μg / mL K2HPO4, 1.0 μg / mL vitamin K), suspending them in the same medium, culturing at 37°C under anaerobic conditions for 24 hours, adding the resulting liquid culture to fresh GAM broth medium at a final concentration of 1 v / v%, culturing under anaerobic conditions at 37°C for 24 hours, centrifuging the resulting culture solution at 13,000 g at 4°C for 15 minutes to remove the cells, and filtering and sterilizing the supernatant fraction with a 0.22 μm filter. To remove the medium components from the culture solution containing the N. mucosa cell culture, after centrifugation at 16,000 g, 4 °C for 30 minutes, the supernatant was discarded, 20 mL of ultrapure water was added, and it was vigorously suspended and washed by vortexing. This washing was repeated 6 times. After washing, centrifugation was performed in the same manner, and the cell culture containing the precipitated cells and EPS was dispersed in ultrapure water and subjected to ultrasonic treatment. For the ultrasonic treatment, BRANSON SONIFIER 150 was used, and ultrasonic treatment was performed under the conditions of power 4, crushing for 20 seconds → cooling for 20 seconds for 5 sets. After the ultrasonic treatment, the cells were precipitated at 16,000 g, 4 °C for 30 minutes, and the supernatant was collected into a new 50 mL tube. To completely remove the cells, the collected supernatant was centrifuged again at 16,000 g, 4 °C for 30 minutes, and the supernatant was collected. By lyophilizing the collected supernatant, the cells and medium components were removed, and a cell culture of N. mucosa containing EPS was obtained.

[0043] (2) Evaluation of infection and infected cells The cell culture of N. mucosa prepared in (1) was dissolved in DMEM / F12 (Gibco) at final concentrations of 0.00625 w / v%, 0.025 w / v%, and 0.1 w / v%, and the inhibitory effect on the infection of Staphylococcus aureus to HeLa cells was evaluated by the same method as in Example 1. The results are shown in Figure 2.

[0044] (4) Results In the evaluation of cell infectivity, a significant decrease in Mean fluorescence intensity (cell infectivity) was observed in the group in which the cell culture of N. mucosa was dissolved (final concentrations of 0.00625 w / v% (0.00625%), 0.025 w / v% (0.025%), and 0.1 w / v% (0.1%)) compared with the control (0) in which the cell culture of N. mucosa was not dissolved (Figure 2).

Claims

1. A topical skin agent for inhibiting Staphylococcus aureus infection, which contains as an active ingredient the cells or cell culture of a bacterium of the genus Neisseria, wherein the bacterium of the genus Neisseria is at least one selected from Neisseria mucosa, Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens and Neisseria elongata, and the cell culture is an EPS-containing culture obtained by adding the culture supernatant of Porphyromonas gingivalis to the culture solution of the bacterium of the genus Neisseria, culturing the mixture, and removing the cells and the medium components.

2. A topical skin agent for preventing or improving skin infections caused by Staphylococcus aureus, which contains as an active ingredient the cells or cell culture of a bacterium of the genus Neisseria, wherein the bacterium of the genus Neisseria is at least one selected from Neisseria mucosa, Neisseria sicca, Neisseria flava, Neisseria subflava, Neisseria flavescens and Neisseria elongata, and the cell culture is an EPS-containing culture obtained by adding the culture supernatant of Porphyromonas gingivalis to the culture solution of the bacterium of the genus Neisseria, culturing the mixture, and removing the cells and the medium components.

3. The agent according to claim 1 or 2, wherein the Staphylococcus aureus is a drug-resistant Staphylococcus aureus.

Images