Beneficial effect of the probiotic bacterium Bacillus subtilis on low-grade inflammation

Abstract

The present invention relates to a strain of Bacillus subtilis, in particular the Bacillus subtilis CU1 strain (CNCM I-2745), or cells obtained by culture of said strain, or a composition comprising said strain or said cells, said composition advantageously being a functional food or a food supplement, for its use in preventing, reducing and / or treating low-grade inflammation, particularly in healthy elderly subjects.

Description

Title of the invention: Beneficial effect of a probiotic bacterium Bacillus subtilis on low-grade inflammation FIELD OF THE INVENTION

[0001] The present invention relates to the field of immunity, in particular the prevention, reduction and / or treatment of low-grade inflammation, especially in the elderly, by a specific probiotic strain of Bacillus subtilis, administered in the form of a functional food or food supplement. STATE OF THE PRIOR ART

[0002] So-called low-grade inflammation is a condition characterized by chronic, low-level systemic inflammation, without major clinical symptoms, creating a general inflammatory background. It is characterized by the expression, in the affected tissues, of a high level of inflammatory factors, in particular circulating cytokines, without presenting any alteration or loss of their primary function.

[0003] The causes of the appearance of low-grade inflammation are multiple and sometimes complicated to determine, but the main ones identified are the following: - Presence of multiple small, unrecognized infectious foci; - Obesity, overweight, metabolic syndrome and insulin resistance; - Intestinal disturbances: dysbiosis and intestinal hyperpermeability; - Lifestyle: sedentary lifestyle, alcohol, stress, tobacco and Western nutrition (sugar, processed products, etc.); - Allergies, intolerances; and - Age.

[0004] Thus, it is accepted that most elderly people develop this type of inflammation, which is then called "inflammageing" (or "inflammaging").

[0005] This inflammation has the consequence of promoting and / or amplifying most of the current chronic diseases, in particular cardiovascular diseases, metabolic diseases (type II diabetes and obesity), cancers, autoimmune diseases, depression and neurological or neuropsychiatric diseases (Alzheimer's, Parkinson's, bipolar disorders, etc.).

[0006] Current management of this inflammation is based on dietary rebalancing, as well as taking food supplements, particularly those based on probiotics.

[0007] Various strains of Bacillus, including Bacillus subtilis (B. subtilis), are currently marketed for use as probiotics in humans and the animal in order to improve their health.

[0008] In particular, the strain of B. subtilis CU1 (or BSCU1), deposited at the CNCM (National Collection of Cultures of Microorganisms, Pasteur Institute, 28 rue du Docteur Roux, 75724 Paris Cedex 15) under number 1-2745 dated October 25, 2001, is known for its use as a beneficial probiotic.

[0009] Document WO2014 / 091160 describes the use of this strain for the prevention and / or treatment of diarrhea.

[0010] Document WO2014 / 207360 describes its use for the prevention and / or treatment of pathologies of the chronic inflammatory rheumatism type, in particular rheumatoid arthritis.

[0011] It should be noted, however, that it is apparent from the state of the art that certain oral compositions based on probiotics do not have a systemic effect given the intestinal mucosa and its barrier function which ensures confinement of these compositions in the intestinal lumen / an absence of absorption and translocation of the luminal contents towards the circulatory system. This absence of systemic effect of compositions of the state of the art is probably due to the choice of strains of bacteria and / or concentrations administered.

[0012] There is a clear need to find solutions to prevent and / or combat low-grade inflammation.

[0013] DESCRIPTION OF THE INVENTION Definitions

[0014] The definitions below correspond to the meaning generally used in the context of the invention and are to be taken into account, unless another definition is explicitly indicated.

[0015] For the purposes of the invention, the articles "a" and "an" are used to refer to one or more (e.g., at least one) units of the grammatical object of the article. For example, "an element" designates at least one element, i.e., one or more elements.

[0016] The terms "about" or "approximately", used in reference to a measurable value such as a quantity, a time, and the like, should be understood to encompass measurement uncertainties of ± 20% or ± 10%, preferably ± 5%, even more preferably ± 1%, and particularly preferably ± 0.1% of the specified value.

[0017] Intervals: Throughout this description, the various features of the invention may be presented in the form of an interval of values. It should be understood that the description of values ​​in the form of an interval is solely for the purpose of making reading easier and should not be interpreted as a rigid limitation of the scope of the invention. Accordingly, the description of an in range of values ​​should be considered to specifically disclose all possible intermediate intervals as well as each of the values ​​within that range. For example, the description of an interval from 1 to 6 should be considered to specifically describe each of the intervals it comprises, such as the intervals from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as each of the values ​​within that range, for example, 1; 2; 2.7; 3; 4; 5; 5.3 and 6. This definition holds regardless of the scope of the interval.

[0018] The term "isolated" should be understood in the context of the invention as synonymous with "removed" or "extracted from its natural environment or state." For example, an isolated strain of bacteria or peptide is a strain of bacteria or peptide extracted from the natural environment in which it is usually found, whether a plant or a living animal, for example. Thus, a strain of bacteria or peptide naturally present in a living animal is not an isolated strain of bacteria or peptide for the purposes of the invention, while the same strain of bacteria or peptide, partially or completely separated from other elements present in its natural context, is "isolated" for the purposes of the invention. An isolated strain of bacteria or peptide may exist in a substantially purified form, or may exist in a non-native environment such as, for example, a host cell.

[0019] Surprisingly, the Applicant has identified a specific probiotic bacterial strain whose use makes it possible to prevent and / or treat low-grade inflammation, particularly in a healthy elderly subject. Thus, it is demonstrated in the present application that this strain has an effect on peripheral innate immunity and that its oral intake results in a reduction in the circulating level of certain pro-inflammatory cytokines.

[0020] The term "probiotic" is used to define live microorganisms which, when integrated in sufficient quantity, exert a positive effect on health, comfort and well-being, beyond traditional nutritional effects.

[0021] Thus and according to a first aspect, the invention relates to the strain of Bacillus subtilis, deposited with the CNCM (National Collection of Cultures of Microorganisms, Pasteur Institute, 28 rue du Docteur Roux, 75724 Paris Cedex 15) on October 25, 2001 under number 1-2745, for use in preventing and / or treating low-grade chronic inflammation.

[0022] In the remainder of the description, the expressions “Bacillus subtilis strain deposited with the CNCM on October 25, 2001 under number 1-2745”, “CNCM 1-2745 strain”, “Bacillus subtilis CU1 strain”, “(probiotic) CU1 strain” and “(probiotic) BSCU1 strain” are used interchangeably.

[0023] In other words, the present invention relates to: - the use of the B. subtilis strain, deposited with the CNCM on October 25, 2001 under number 1-2745, to prevent and / or treat low-grade chronic inflammation; - the use of the strain of B. subtilis, deposited with the CNCM on October 25, 2001 under number 1-2745, to prepare a medicament, advantageously a functional food, in particular a food supplement, to prevent and / or treat low-grade chronic inflammation; - a method for preventing and / or treating low-grade chronic inflammation in a subject comprising the administration, advantageously orally, of the strain of B. subtilis, deposited with the CNCM on October 25, 2001 under number I-2745.

[0024] Low-grade inflammation is a condition called "chronic low-grade inflammation," as opposed to acute inflammation, which is a pathology.

[0025] As known to those skilled in the art, the first difference is based on the duration of the inflammation, which, in the case of acute inflammation, will be short-lived, whereas low-grade inflammation is a so-called “chronic” inflammation that develops slowly.

[0026] The second difference lies in the clinical signs of inflammation: acute inflammation is characterized by 4 major clinical signs: redness, heat, edema and pain, whereas low-grade inflammation is silent, without any direct and clearly identifiable clinical signs.

[0027] Furthermore, in acute inflammation, the vascular and cellular phases follow one another with a resolving phase. In low-grade inflammation, the first two phases (vascular and cellular) coexist and there is no resolving phase.

[0028] A discriminating marker is the "C-reactive protein" (or CRP), synthesized by hepatocytes during an inflammatory reaction under the influence of different mediators themselves involved in inflammation, notably cytokines. In the case of acute inflammation, the blood level of CRP appears in the form of a peak, with values ​​between 20 and 500 mg / L, knowing that a physiological level must be below the 6 mg / L mark. During low-grade inflammation, CRP, to be measured using an ultrasensitive method (ultrasensitive CRP or CRP-us), is around 2 to 5 mg / L, i.e. a low but constant level.

[0029] Note that low-grade inflammation is a chronic inflammatory condition which is, however, distinguished from a chronic inflammatory disease / pathology by the absence of symptoms, or even lower CRP levels.

[0030] As highlighted in the present application, the specific strain targeted has the ability to decrease chronic low-grade inflammation. In the context of the application, the terms "decrease", "reduce" or "attenuate" are used interchangeably interchangeable. This includes, in particular, the decrease in the level of blood markers, notably circulating cytokines, particularly detected in the serum.

[0031] According to a particular embodiment, the cytokines whose level is reduced by the administration of the strain targeted in the application are chosen from the following group: IL-8 (Interleukin 8), TNF-a (alpha Tumor Necrosis Factor), and CCL3 (Chemokine (CC motif) Ligand 3 or Macrophage Inflammatory Protein 1-alpha (MIP-1-a)).

[0032] According to one embodiment, the level of these cytokines is reduced by at least 5%, or even 10%, or even 15% or 20%, in particular after taking the strain B. subtilis CNCM 1-2745, preferably for at least 4 weeks, advantageously in a daily quantity of 2 x 109 CFU (for “Colony Forming Unit”).

[0033] According to an advantageous embodiment, such a reduction is observed at least for the cytokines IL-8, TNF-a and CCL3.

[0034] In other words, the specific strain targeted is intended to prevent and / or treat low-grade chronic inflammation.

[0035] By "preventing a condition", here low-grade inflammation, we mean avoiding its occurrence, particularly in subjects at risk of developing it, or delaying its repercussions, in this case by avoiding the increase in inflammatory intensity likely to lead to other pathologies. In this context, we speak of preventive or prophylactic treatment of this low-grade inflammatory condition.

[0036] In particular, the term "prevent" means administering such a strain before the onset of this condition, for example when the levels of IL-8, TNF-a, CCL3 are at a so-called normal level.

[0037] Alternatively, "preventing" means administering such a strain before the onset of this condition, for example when the CRP level is less than 2 mg / L.

[0038] Since low-grade chronic inflammation is very common in the elderly, particularly those aged 65 and over, it may be recommended to administer this strain to subjects under 65, or even under 60 or even 55.

[0039] By "treating a condition", here low-grade inflammation, we mean improving it, relieving its repercussions, in this case by reducing the inflammatory intensity likely to lead to other pathologies, or at least stabilizing it. In this context, we speak of curative treatment of this low-grade inflammatory condition.

[0040] In particular, the term “treating” or “fighting against” means administering such a strain when this condition is established, for example when the levels of IL-8, TNF-a, CCL3 are at a level higher than a so-called normal level, advantageously as soon as that an increase in this level is observed.

[0041] Alternatively, "treating" or "fighting" means administering such a strain when this condition is established, for example when the CRP level is in the order of 2 to 5 mg / L.

[0042] Since low-grade chronic inflammation is very widespread in the elderly, particularly those aged 65 and over, it may be recommended to administer this strain systematically to elderly subjects, for example those over 65.

[0043] As already mentioned, low-grade inflammation is known to promote and / or amplify most current chronic diseases, in particular cardiovascular diseases, metabolic diseases (type II diabetes and obesity), cancers, autoimmune diseases, depression and neurological or neuropsychiatric diseases (Alzheimer's, Parkinson's, bipolar disorders, etc.). Therefore, preventing and / or treating low-grade inflammation makes it possible to avoid or delay the onset of these diseases, or even to alleviate them.

[0044] According to a preferred embodiment, the strain used within the framework of the invention is intended for human and / or veterinary use, advantageously for human use.

[0045] According to an advantageous embodiment, the strain used in the context of the invention is intended for an elderly subject. In the context of the invention, the term "elderly subject" means a man or a woman, over 55 years old, or even over 60 years old, or even 65 years old or more.

[0046] According to a particular embodiment, the subject receiving the administration of the strain used in the context of the invention has at least one or even all of the following characteristics: - age between 65 and 79 years; - body mass index (BMI) between 22 and 28 kg / m2; - good general condition; - non-smoker.

[0047] According to one embodiment, the strain used in the context of the invention is intended for a so-called healthy subject. In the context of the invention, the term "healthy subject" means a healthy man or woman, in whom no pathology or disease has been diagnosed and / or declared, in particular chosen from the following group: cardiovascular diseases, metabolic diseases, in particular type II diabetes and obesity, cancers, autoimmune diseases, depression, and neurological or neuropsychiatric diseases such as Alzheimer's, Parkinson's and bipolar disorders.

[0048] Alternatively, the strain used in the context of the invention may be intended for a subject suffering from a disease or condition chosen from the following group: cardiovascular diseases, metabolic diseases, in particular type II diabetes and obesity, cancers, depression, and neurological or neuropsychiatric diseases such as Alzheimer's, Parkinson's and bipolar disorders.

[0049] According to another embodiment and at the time of administration of the strain used in the context of the invention, the subject does not exhibit one or all of the following characteristics: - a chronic illness, e.g., diabetes mellitus, congestive heart failure, respiratory failure, cancer, chronic kidney or liver disease; - acute infection in the previous month; - gastrointestinal disease, e.g. inflammatory bowel disease; - immune disorder; - immunodeficiency; - autoimmune diseases; - acute gastroenteritis in the last 2 months; - vaccination or taking antibiotics, anti-inflammatories, or immunosuppressants at the time of administration; - regular use of laxatives; - taking drugs or excessive alcohol; - chronic inflammatory rheumatism, particularly rheumatoid arthritis.

[0050] According to one embodiment, the subject receiving the administration of the strain used in the context of the invention has a blood CRP level of less than 6 mg / L, in particular between 2 and 5 mg / L.

[0051] According to another embodiment, the subject receiving the administration of the strain used in the context of the invention has at least one of the following characteristics: - Presence of multiple small, unrecognized infectious foci; - Obesity, overweight, metabolic syndrome and insulin resistance; - Intestinal disturbances: dysbiosis and intestinal hyperpermeability; - Sedentary lifestyle, alcohol, stress, tobacco and / or Western nutrition (sugar, processed products, etc.); - Allergies and / or intolerances; - CRP, advantageously measured using an ultrasensitive method (ultrasensitive CRP or CRP-us), less than 6 mg / L or even between 2 and 5 mg / L, in this case a relatively low and constant blood level.

[0052] According to another aspect, the invention relates to cells obtained by culturing the strain of Bacillus subtilis, deposited with the CNCM (National Collection of Cultures of Microorganisms, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15) on October 25, 2001 under number 1-2745, for use in preventing and / or treating low-grade chronic inflammation.

[0053] In other words, the present invention relates to: - the use of cells obtained by culturing the B. subtilis strain, deposited with the CNCM on October 25, 2001 under number 1-2745, to prevent and / or treat low-grade chronic inflammation; - the use of cells obtained by culturing the B. subtilis strain, filed with the CNCM on October 25, 2001 under number 1-2745, to prepare a medicament, advantageously a functional food, in particular a food supplement, to prevent and / or treat low-grade chronic inflammation; - a method for preventing and / or treating low-grade chronic inflammation in a subject comprising the administration, advantageously orally, of cells obtained by culturing the strain of B. subtilis, filed with the CNCM on October 25, 2001 under number 1-2745.

[0054] In practice, the cells are obtained by culturing the bacterial strain in a culture medium, according to methods well known to those skilled in the art, for example according to those described in the work “Biotechnology, 5th edition, R. Scriban, Edition Tec. & Doc., 1999, ISBN: 2-7430-0309-X”.

[0055] Typically, a method for producing cells by culturing the strain implemented within the framework of the invention comprises the following steps: - inoculation of a culture medium with an inoculum of the strain; - culture in aerobic conditions, to obtain cell multiplication; - separation of the biomass from its culture medium, to obtain cells.

[0056] The cells thus obtained are predominantly in vegetative form. The “vegetative form” of a bacterium designates the form of a bacterium placed in favorable conditions. The expression “predominantly in vegetative form” means that at least 70% of the cells are in vegetative form, preferably at least 80%, and more preferably at least 90%. An example of favorable conditions is a non-limiting culture medium at a temperature and pH favorable to bacterial multiplication. A non-limiting culture medium contains all the nutrients necessary for cell multiplication.

[0057] The cell production method may further comprise an intermediate step, between the aerobic culture step and the step of separating the biomass from its culture medium, of placing the cells in unfavorable conditions. The cells thus obtained are then mainly in sporulated form.

[0058] The "spore-forming form" of a bacterium refers to the form of a bacterium placed in unfavorable conditions. The expression "predominantly in spore-forming form" means that at least 70% of the cells are in spore-forming form, preferably at least 80%, and more preferably at least 90%. The spore-forming form is a form of re resistance that allows cells to withstand a hostile environment such as a lack of nutrients, i.e. a limiting nutrient environment, water stress, a significant variation in pH or temperature, or even crossing the digestive tract.

[0059] Placing the cells in unfavorable conditions is, for example, obtained by: not renewing the bacteria culture medium, stopping the supply of culture medium, using a limiting culture medium, changing the temperature and / or pH, controlling the aeration and / or agitation so as to maintain an O2 pressure greater than 20% in the culture medium and a CO2 pressure less than 1.5% in the outgoing gases or their combination. It is also possible to obtain cells predominantly in sporulated form by adding glutamic acid to the culture medium in an amount of around 0.75g / L.

[0060] In a preferred embodiment, the cells are predominantly in sporulated form.

[0061] The cell production method may also comprise a subsequent step of drying the cells to obtain cells in dry form. The drying is, for example, freeze-drying, fluidized bed drying or spray drying.

[0062] In a particular embodiment, the cells are in dry form. “Cells in dry form” advantageously means that the biomass obtained at the end of the cell production process advantageously comprises more than 90% dry matter, preferably more than 95% dry matter.

[0063] According to another aspect, the invention relates to a composition comprising the strain of Bacillus subtilis, deposited with the CNCM (National Collection of Cultures of Microorganisms, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15) on October 25, 2001 under number 1-2745, or cells obtained by culturing said strain, for use in preventing and / or treating low-grade chronic inflammation.

[0064] In other words, the present invention relates to: - the use of a composition comprising the strain of B. subtilis, deposited with the CNCM on October 25, 2001 under number 1-2745, or cells obtained by culturing said strain, to prevent and / or treat low-grade chronic inflammation; - the use of a composition comprising the strain of B. subtilis, deposited with the CNCM on October 25, 2001 under number 1-2745, or cells obtained by culturing said strain, to prepare a medicament, advantageously a functional food, in particular a food supplement, to prevent and / or treat low-grade chronic inflammation; - a method for preventing and / or treating chronic low-grade inflammation in a subject comprising the administration, advantageously orally, of a composition comprising the strain of B. subtilis, deposited with the CNCM on October 25, 2001 under number 1-2745, or cells obtained by culture of said strain.

[0065] A composition used within the scope of the invention may be a pharmaceutical composition, which may contain pharmaceutically acceptable excipients, or a nutraceutical composition, which may contain nutraceutically acceptable excipients.

[0066] According to a preferred embodiment, the composition used in the context of the invention is a nutraceutical composition, including a product comprising a food additive (functional food), or a food supplement. Nutraceutically acceptable excipients are, for example, maltodextrin and magnesium stearate.

[0067] A food product can be any type of food, drink or confectionery. It can for example be a drink, a cereal bar, chewing gum, a dairy product, possibly fermented, to which the strain used in the context of the present invention is added.

[0068] By definition, a food supplement is defined as a foodstuff whose purpose is to supplement the normal diet and which constitutes a concentrated source of nutrients or other substances having a nutritional or physiological effect, alone or in combination.

[0069] In practice, a food supplement is marketed in dose form, for example with presentation forms such as capsule, lozenge, tablet, pill and other similar forms, sachet of powder, ampoule of liquid, bottle equipped with a dropper and other similar forms of liquid or powder preparations intended to be taken in measured units of small quantity.

[0070] In a known manner, the composition used in the context of the invention may comprise the usual adjuvants or excipients used in the field in question, such as hydrophilic or lipophilic thickeners or gelling agents, appetizers, hydrophilic or lipophilic additives, preservatives, antioxidants, diluents, vitamins, minerals, suspending agents, cellulose derivatives, absorbents, cryoprotective agents or even colorants.

[0071] Of course, the person skilled in the art will take care to choose this or these possible adjuvants or excipients, and to adjust their quantity, in such a way that the advantageous properties of the composition according to the invention are not, or not substantially, altered by the envisaged addition.

[0072] According to one embodiment, the composition according to the invention comprises nutritive substances usable as a support and / or prebiotic substances. advantageously chosen from fructo-oligosaccharides, inulins, isomalto-oligosaccharides, lactitol, lactosaccharose, lactulose, pyrodextrins, soy oligosaccharides, transgalacto-oligosaccharides, xylo-oligosaccharides, yeast walls, vitamins, in particular vitamin E.

[0073] According to another embodiment, the composition according to the invention comprises at least one compound chosen from the following group: zeolites, calcium carbonate, calcium sulfate, magnesium carbonate, talc, trehalose, chitosan, shellac, albumin, starch, skimmed milk powder, whey, whey powder, maltodextrins, lactose, inulin, dextroses, celluloses, clays including sepiolite, yeast and cereal derivatives, vegetable oils or a solvent chosen from water or a physiological solution.

[0074] According to a particular embodiment, the composition according to the invention comprises a coating material advantageously chosen from maltodextrins, guar seed flour, gum arabic, alginates, modified starch and starch derivatives, dextrins, cellulose derivatives such as cellulose ester and cellulose ether, proteins such as gelatin, albumin, casein, gluten, gum arabic, gum tragacanth, lipids such as waxes, paraffin, stearic acid, mono- and diglycerides.

[0075] The mixture or composition according to the invention may be in the form of a powder, a capsule, a spray, a solution, an emulsion, a suspension or a dispersion.

[0076] Advantageously, the mixture or composition according to the invention is in dry or liquid form, in particular in lyophilized, dried, pressed, liquid or frozen form, advantageously in lyophilized form.

[0077] According to another preferred embodiment, the composition used in the context of the invention is in a form suitable for oral administration, whether solid or liquid. Thus, such a composition may be in the form of a capsule, pill, lozenge, tablet, capsule, powder, suspension, solution, syrup or granule.

[0078] According to another embodiment, the composition used in the context of the invention is devoid of any other active ingredient, advantageously of any other microorganism, in particular of any other probiotic, in particular of bacterial origin. In other words, the composition may contain only, as living microorganism, the strain targeted in the present invention.

[0079] Alternatively, the strain targeted by the invention can be combined with other microorganisms, advantageously probiotics, in particular of bacterial origin such as the genera Bacillus, Bifidobacterium, Lactobacillus, Lactococcus, Lactiplanti-bacillus, Ligilactobacillus and Bifidobacterium, or yeasts such as Sac-charomyces, Kluyveromyces, Pichia and Torulaspora.

[0080] Suitably, a composition according to the invention contains a quantity of cells corresponding to the daily dose recommended for the intended application.

[0081] Note that the daily dose may depend on the type of subject, its weight, the method of administration and the type of curative or preventive treatment. Thus, such a daily quantity may range from 105 to 1011 CFU (for “Colony Forming Unit”), for example 105, 106, 107, 108, 109, 1010, 1011 CFU per day.

[0082] Suitably for human use, a composition used in the context of the invention comprises from 1.108 to 1.1011 CFU, advantageously from 1.109 CFU to 1.1010 CFU, for example 2.109, 3.109, 4.109, 5.109, 6.109, 7.109, 8.109 or 9.109 CFU, even more advantageously 2.109 CFU.

[0083] According to one embodiment, the strain, cells or composition used in the context of the present invention are administered daily, advantageously once or twice a day, in particular before taking a meal, for example breakfast.

[0084] According to another embodiment, the strain, cells or composition used in the context of the present invention are administered continuously and regularly for several weeks, namely at least one week, or even 2, 3 or 4 weeks, or even at least 1 month or more (several months to several years).

[0085] Thus and according to a particular embodiment, the daily intake of the preventive or curative treatment is done without interruption and / or in a non-sequential manner (for example successive cycles of 10 days of intake, followed by 18 days of break).

[0086] The manner in which the invention can be implemented and the advantages which result therefrom will become more apparent from the following examples of implementation, given for informational and non-limiting purposes, in support of the appended figure. FIGURES

[0087] [Fig.l]: Cytokine levels (IL-8, TNF-a and CCL3) quantified in the serum of healthy elderly subjects (65 < age < 79 years) before administration of the probiotic composition of the invention (“Baseline”) then 4 weeks after daily administration (“Week 4”).

[0088] Data represent mean values ​​+ / - SE (“Standard Error”) EXAMPLES OF ACHIEVEMENT

[0089] All experiments were carried out using the B. subtilis CU1 strain deposited on October 25, 2001 at the CNCM (National Collection of Cultures of Microorganisms, Pasteur Institute, 28 rue du Docteur Roux, 75724 Paris Cedex 15) under number 1-2745. 1. Materials and methods 1.1. Populations

[0090] The subjects (30 people) were recruited from the healthy general population.

[0091] The study was a single-center trial.

[0092] To participate in this study, subjects had to meet all of the following criteria: - 65 < age < 79 years; - 22.0 < BMI < 28.0 kg / m2; and - Non-smoking 1.2. Administration

[0093] The probiotic composition according to the invention comprises BSCU1 cells mixed with excipients (maltodextrin and magnesium stearate; see Table 1). This composition corresponds to the product LifeinU® BSCU1 marketed by GNOSIS by LESAFFRE (Marcq en Baroeul, France).

[0094] Each probiotic capsule contains 2 x 109 CFU of B. subtilis, the majority of which are in sporulated form (i.e., B. subtilis CU1 spores).

[0095] The capsules were taken orally, as a dietary supplement.

[0096] The subjects were instructed to consume one capsule of the study product daily for 4 weeks, in the morning (before breakfast), i.e. a daily dose of 2 x 109 CFU of B. subtilis CU1 (CNCM 1-2745).

[0097] [Tables 1] Ingredients Quantity (mg) Quantity (%) LifeinU™ BSCU1 (BSCU1 CNCM 1-2745) 20 6.67 Maltodextrin glucidex 19 (Arugula) 270 90 Magnesium stearate 10 3.33 Total 300 100 1.3. Analysis of cytokines in plasma

[0098] A blood sample was taken before administration of the probiotic composition of the invention and then 4 weeks after this daily administration and used within the following hour.

[0099] BD Vacutaainer CAT 2 mL PET tubes (BD Biosciences, Vianen, The Netherlands, BD 368271) were used for serum preparation. Samples were incubated at room temperature and centrifuged at 2000xg for 10 minutes. Sera were collected and stored at -80°C for cytokine analysis.

[0100] Cytokine concentration was determined using the Bio-Plex Pro™ Human Cytokine Assay (BioRad, US) according to the manufacturer's instructions. Fluorescence was measured using the Bio-Plex 200 Array Reader and Bio-Plex Manager 6.1 software. Concentrations were calculated using a standard curve. The following cytokines were analyzed: IL-8, CCL3, and TNF-a. All cytokines were within the standard range. 2. Results

[0101] The effect of 4 weeks of daily BSCU1 supplementation on low-grade inflammation in an elderly group is illustrated in [Fig. 1].

[0102] It was found that plasma levels of IL-8, TNF-a and CCL3 decreased by 21%, 21% and 17%, respectively, after treatment with BSCU1.

[0103] These results clearly show that the consumption of BSCU1 contributes to reducing chronic low-grade inflammation in this population.

Claims

Claims

1. Strain of Bacillus subtilis, deposited with the CNCM (National Collection of Cultures of Microorganisms, Institut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15) on October 25, 2001 under number 1-2745, for use in preventing and / or treating low-grade chronic inflammation.

2. Cells obtained by culturing the strain of B. subtilis, deposited with the CNCM on October 25, 2001 under number 1-2745 for use in preventing and / or treating low-grade inflammation.

3. Cells for their use according to claim 2, characterized in that they are in sporulated and / or vegetative form, advantageously sporulated, preferably sporulated and freeze-dried.

4. Composition comprising the strain of Bacillus subtilis deposited with the CNCM on October 25, 2001 under number 1-2745 or cells obtained by culturing said strain for use in preventing and / or treating low-grade inflammation in an elderly subject.

5. Composition for its use according to claim 4, characterized in that it comprises the strain or cells in an amount greater than or equal to 1.109 CFU, advantageously between 1.109 and 1.1010 CFU, for example 2.109 CFU.

6. Composition for its use according to one of claims 4 to 5, characterized in that it is in a form suitable for oral administration, advantageously in the form of a capsule, pill, lozenge, tablet, capsule, powder, suspension, solution or granule.

7. Composition for its use according to one of claims 4 to 6, characterized in that said composition is a functional food or a food supplement.

8. Strain for its use according to claim 1, cells for their use according to one of claims 2 to 3, or composition for its use according to one of claims 4 to 7 characterized in that the subject is elderly.

9. Strain for its use according to claim 1 or 8, cells for their use according to one of claims 2 to 3 or 8, or composition for its use according to one of claims 4 to 8 characterized in that the subject is aged: - less than 65 years, advantageously less than 60 years, preferably less than 55 years; or

10. - at least 60 years old, advantageously at least 65 years old. Strain for its use according to claim 1, 8 or 9, cells for their use according to one of claims 2 to 3, 8 or 9, or composition for its use according to one of claims 4 to 9 characterized in that the subject is healthy.