COMPOSITIONS CONTAINING BACTERIAL STRAINS

Abstract

The invention relates to compositions comprising bacterial strains for the treatment and prevention of inflammatory and autoimmune diseases.

Description

TECHNICAL FIELD

[0001] This invention is in the field of compositions comprising bacterial strains isolated from the mammalian digestive tract and the use of such compositions in the treatment of disease.BACKGROUND TO THE INVENTION

[0002] The human intestine is thought to be sterile in utero, but it is exposed to a large variety of maternal and environmental microbes immediately after birth. Thereafter, a dynamic period of microbial colonization and succession occurs, which is influenced by factors such as delivery mode, environment, diet and host genotype, all of which impact upon the composition of the gut microbiota, particularly during early life. Subsequently, the microbiota stabilizes and becomes adult-like [1]. The human gut microbiota contains more than 500-1000 different phylotypes belonging essentially to two major bacterial divisions, the Bacteroidetes and the Firmicutes [2]. The successful symbiotic relationships arising from bacterial colonization of the human gut have yielded a wide variety of metabolic, structural, protective and other beneficial functions. The enhanced metabolic activities of the colonized gut ensure that otherwise indigestible dietary components are degraded with release of by-products providing an important nutrient source for the host. Similarly, the immunological importance of the gut microbiota is well-recognized and is exemplified in germfree animals which have an impaired immune system that is functionally reconstituted following the introduction of commensal bacteria [3-5].

[0003] Dramatic changes in microbiota composition have been documented in gastrointestinal disorders such as inflammatory bowel disease (IBD). For example, the levels of Clostridium cluster XIVa bacteria are reduced in IBD patients whilst numbers of E. coli are increased, suggesting a shift in the balance of symbionts and pathobionts within the gut [6-9]. Interestingly, this microbial dysbiosis is also associated with imbalances in T effector cell populations.

[0004] In recognition of the potential positive effect that certain bacterial strains may have on the animal gut, various strains have been proposed for use in the treatment of various diseases (see, for example, [10-13]). Also, certain strains, including mostly Lactobacillus and Bifidobacterium strains, have been proposed for use in treating various inflammatory and autoimmune diseases that are not directly linked to the intestines (see

[14] and

[15] for reviews). However, the relationship between different diseases and different bacterial strains, and the precise effects of particular bacterial strains on the gut and at a systemic level and on any particular types of diseases, are poorly characterised.

[0005] WO2011 / 110918 and WO2011 / 149335 describe clinical studies using probiotics in the treatment of atopic dermatitis and allergies.WO2009 / 072889 describes clinical studies using probiotics in the treatment of allergic asthma.

[0006] There is a requirement in the art for new methods of treating inflammatory and autoimmune diseases. There is also a requirement for the potential effects of gut bacteria to be characterised so that new therapies using gut bacteria can be developed.SUMMARY OF THE INVENTION

[0007] The invention provides a composition comprising a bacterial strain having a genome with at least 90% sequence identity to SEQ ID NO: 2 of PCT / GB2016 / 051776, for use in a method of treating or preventing allergic asthma, eosinophilic asthma or airway hyper-responsiveness associated with asthma.

[0008] The inventors have developed new therapies for treating and preventing inflammatory and autoimmune diseases. In particular, the inventors have developed new therapies for treating and preventing diseases and conditions mediated by IL-17 or the Th17 pathway. In particular, the inventors have identified a new bacterial strain that is effective for reducing the Th17 inflammatory response. As described in the examples, oral administration of compositions comprising the bacterium deposited under accession number NCIMB 42380 may reduce the severity of the inflammatory response, including the Th17 inflammatory response, in mouse models of asthma, rheumatoid arthritis and multiple sclerosis. As also described in the examples, oral administration of compositions comprising the bacterium deposited under accession number NCIMB 42380 may reduce tumor size in mouse models of cancer that may be associated with the Th17 inflammatory response.

[0009] The inventors have identified that treatment with such bacterial strains can reduce levels of cytokines that are part of the Th17 pathway, including IL-17, can alleviate the Th17 inflammatory response and can provide clinical benefits in mouse models of inflammatory and autoimmune diseases mediated by IL-17 and the Th17 pathway.

[0010] In preferred embodiments, the invention provides a composition comprising the bacterium deposited under accession number NCIMB 42380, for use in a method of treating or preventing allergic asthma. The inventors have identified that treatment with the bacterium deposited under accession number NCIMB 42380 can reduce recruitment of neutrophils and eosinophils into the lungs, which can help treat or prevent asthma. Furthermore, the inventors have tested and demonstrated the efficacy of the bacterium deposited under accession number NCIMB 42380 in mouse models of asthma. In certain embodiments, the composition is for use in a method of treating or preventing eosinophilic asthma. The effect shown for the compositions of the invention on eosinophils mean that they may be particularly effective for treating or preventing eosinophilic asthma. Indeed, in certain embodiments, the composition is for use in a method of reducing an eosinophilic inflammatory response in the treatment or prevention of eosinophilic asthma. The bacterium deposited under accession number NCIMB 42380 is shown to have a particularly pronounced effect on neutrophils in asthma models and treatment with this bacterium may be particularly effective for treating neutrophilic asthma.

[0011] In particular, the compositions of the invention may be used in reducing IL-17 production or reducing Th17 cell differentiation in the treatment or prevention of allergic or eosinophilic asthma.

[0012] In certain embodiments, the composition is for use in a patient with elevated IL-17 levels or Th17 cells. The effect on the Th17 inflammatory response shown for the bacterium deposited under accession number NCIMB 42380 may be particularly beneficial for such patients.

[0013] In preferred embodiments of the invention, the bacterial strain in the composition is the bacterium deposited under accession number NCIMB 42380. The bacterial strains may have a 16s rRNA sequence that is at least 99%, 99.5% or 99.9% identical to SEQ

[0014] ID NO:1. Preferably, the bacterial strain for use in the invention has the 16s rRNA sequence represented by SEQ ID NO:1.

[0015] In certain embodiments, the composition of the invention is for oral administration. Also, oral administration is convenient for patients and practitioners and allows delivery to and / or partial or total colonisation of the intestine.

[0016] In certain embodiments, the composition of the invention comprises one or more pharmaceutically acceptable excipients or carriers.

[0017] In certain embodiments, the composition of the invention comprises a bacterial strain that has been lyophilised. Lyophilisation is an effective and convenient technique for preparing stable compositions that allow delivery of bacteria.

[0018] In certain embodiments, the invention provides a food product comprising the composition as described above.

[0019] In certain embodiments, the invention provides a vaccine composition comprising the composition as described above.

[0020] In developing the above invention, the inventors have identified and characterised a bacterial strain that is particularly useful for therapy. The bacterium deposited under accession number NCIMB 42380 is shown to be effective for treating the diseases described herein, such as arthritis, asthma and multiple sclerosis. The bacterium deposited under accession number NCIMB 42380 is also shown to be effective for treating cancer.BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1: Mouse model of house dust mite-induced asthma - Total BAL fluid cell counts. Figure 2: Mouse model of house dust mite-induced asthma - Total eosinophil count in BALF. Figure 3: Mouse model of house dust mite-induced asthma - Proportion of eosinophils in BALF. Figure 4: Mouse model of house dust mite-induced asthma - Total macrophage count in BALF. Figure 5: Mouse model of house dust mite-induced asthma - Proportion of macrophages in BALF. Figure 6: Mouse model of house dust mite-induced asthma - Total neutrophil count in BALF. Figure 7: Mouse model of house dust mite-induced asthma - Proportion of neutrophils in BALF. Figure 8: Mouse model of house dust mite-induced asthma - Total lymphocyte count in BALF. Figure 9: Mouse model of house dust mite-induced asthma - Proportion of lymphocytes in BALF. Figure 10: Mouse model of severe neutrophilic asthma - Total BAL fluid cell counts. Figure 11: Mouse model of severe neutrophilic asthma - Total eosinophil count in BALF. Figure 12: Mouse model of severe neutrophilic asthma - Proportion of eosinophils in BALF. Figure 13: Mouse model of severe neutrophilic asthma - Total macrophage count in BALF. Figure 14: Mouse model of severe neutrophilic asthma - Proportion of macrophages in BALF. Figure 15: Mouse model of severe neutrophilic asthma - Total neutrophil count in BALF. Figure 16: Mouse model of severe neutrophilic asthma - Proportion of neutrophils in BALF. Figure 17: Mouse model of severe neutrophilic asthma - Total lymphocyte count in BALF. Figure 18: Mouse model of severe neutrophilic asthma - Proportion of lymphocytes in BALF. Figure 19: Mouse model of rheumatoid arthritis - Bodyweights, days -14 to 0. Data are presented as Mean ± SEM percentages of the initial (Day -14) bodyweights. Statistical significance: ▲ p < 0.05 and ▲▲▲▲ p < 0.0001 when compared to the vehicle-treated group. Figure 20: Mouse model of rheumatoid arthritis - Bodyweights, days 0 to 42. Data are presented as Mean ± SEM percentages of the initial (Day 0) bodyweights. ▲ p < 0.05, ◆ p < 0.05, ▲▲▲ p < 0.001, •••• p < 0.0001 when compared to the vehicle-treated group. Figure 21: Mouse model of rheumatoid arthritis - Clinical Scores. Data are presented as Mean ± SEM. ∗∗∗∗< p < 0.0001 when compared to Day 21 in the vehicle-treated group. ◆,O p < 0.05 when compared to the vehicle-treated group on a given day. Figure 22: Mouse model of rheumatoid arthritis - Splenocyte proliferative response to Collagen II. Media background subtracted [CII-stimulated - media background] counts per minute based on 3H-TdR incorporation. All data are presented as Mean ± SEM. ∗∗< p < 0.01 compared to Vehicle group. Figure 23: Mouse model of rheumatoid arthritis - Levels of IFNγ in tissue culture supernatants. Lines represent group median values. Figure 24: Mouse model of rheumatoid arthritis - Levels of IL-17A in tissue culture supernatants. Lines represent group median values. Figure 25: Mouse model of rheumatoid arthritis - Levels of IL-10 in tissue culture supernatants. Lines represent group median values. Figure 26: Mouse model of rheumatoid arthritis - Levels of IL-6 in tissue culture supernatants. Lines represent group median values. Figure 27: Histopathology scoring system. Figure 28: Mouse model of house dust mite-induced asthma - Total IgE in Serum Figure 29: Mouse model of house dust mite-induced asthma - HDM specific IgG1 in Serum Figure 30: Mouse model of house dust mite-induced asthma - Total IgE in BALF Figure 31: Mouse model of house dust mite-induced asthma - HDM specific IgG1 in BALF Figure 32: Mouse model of house dust mite-induced asthma - Histological Analysis - Mean Peribronchiolar Infiltration Score Figure 33: Mouse model of house dust mite-induced asthma - Histological Analysis - Mean Perivascular Infiltration Score Figure 34: Mouse model of house dust mite-induced asthma - Histological Analysis - Mean Inflammatory Score (Average of both Peribronchiolar and Perivascular Infiltration Score) Figure 35: Mouse model of house dust mite-induced asthma - Histological Analysis - Mucus Score Figure 36: Mouse model of house dust mite-induced asthma - IL-9 level in lung tissue Figure 37: Mouse model of house dust mite-induced asthma - IL-1a level in lung tissue Figure 38: Mouse model of house dust mite-induced asthma - IFNg level in lung tissue Figure 39: Mouse model of house dust mite-induced asthma - IL-17A level in lung tissue Figure 40: Mouse model of house dust mite-induced asthma - IL-4 level in lung tissue Figure 41: Mouse model of house dust mite-induced asthma - IL-5 level in lung tissue Figure 42: Mouse model of house dust mite-induced asthma - IL-1b level in lung tissue Figure 43: Mouse model of house dust mite-induced asthma - RANTES level in lung tissue Figure 44: Mouse model of house dust mite-induced asthma - MIP-1a level in lung tissue Figure 45: Mouse model of house dust mite-induced asthma - KC level in lung tissue Figure 46: Mouse model of house dust mite-induced asthma - MIP-2 level in lung tissue Figure 47: Mouse model of severe neutrophilic asthma - HDM specific IgG1 in Serum Figure 48: Mouse model of severe neutrophilic asthma - HDM specific IgG2a in Serum Figure 49: Mouse model of severe neutrophilic asthma - HDM specific IgG1 in BALF Figure 50: Mouse model of severe neutrophilic asthma - HDM specific IgG2a in BALF Figure 51: Mouse model of severe neutrophilic asthma - Histological Analysis - Mean Peribronchiolar Infiltration Score Figure 52: Mouse model of severe neutrophilic asthma - Histological Analysis - Mean Perivascular Infiltration Score Figure 53: Mouse model of severe neutrophilic asthma - Histological Analysis - Mean Inflammatory Score (Average of both Peribronchiolar and Perivascular Infiltration Score) Figure 54: Mouse model of severe neutrophilic asthma - TNFa level in lung tissue Figure 55: Mouse model of severe neutrophilic asthma - IL-1a level in lung tissue Figure 56: Mouse model of severe neutrophilic asthma - IFNg level in lung tissue Figure 57: Mouse model of severe neutrophilic asthma - IL-17F level in lung tissue Figure 58: Mouse model of severe neutrophilic asthma - IL-1b level in lung tissue Figure 59: Mouse model of severe neutrophilic asthma - RANTES level in lung tissue Figure 60: Mouse model of severe neutrophilic asthma - MIP-2 level in lung tissue Figure 61: Mouse model of severe neutrophilic asthma - KC level in lung tissue Figure 62: Mouse model of severe neutrophilic asthma - IL-17A level in lung tissue Figure 63: Mouse model of severe neutrophilic asthma -MIP-1a level in lung tissue Figure 64: Mouse model of severe neutrophilic asthma - IL-33 level in lung tissue Figure 65: Mouse model of rheumatoid arthritis - Visual Template for Histopathology Scoring. Representative images showing composite scores from mouse tarsal joints in a collagen-induced arthritis study. Figure 66: Mouse model of rheumatoid arthritis - Histopathology: Inflammation Scores. Data are presented as Mean ± SEM. ∗∗< p < 0.01 when compared to the vehicle-treated group. Figure 67: Mouse model of rheumatoid arthritis - Histopathology: Cartilage Scores. Data are presented as Mean ± SEM. ∗∗∗< p < 0.001 when compared to the vehicle-treated group. Figure 68: Mouse model of rheumatoid arthritis - Histopathology: Bone Scores. Data are presented as Mean ± SEM. ∗∗∗< p < 0.001 when compared to the vehicle-treated group. Figure 69: Mouse model of rheumatoid arthritis - Histopathology: Total Scores. Data are presented as Mean ± SEM. ∗< p < 0.05, ∗∗∗< p < 0.001 when compared to the vehicle-treated group. Figure 70: Mouse model of rheumatoid arthritis - Histopathology: Representative Pictures. Animal ID (#n.n) and limb (R for right, L for left) are indicated between brackets. Top left image (vehicle): extensive joint and bone destruction with inflammation and fibrosis extending to the peri-articular soft tissues. Figure 71: Mouse model of multiple sclerosis - clinical score. Figure 72: Mouse model of multiple sclerosis - disease incidence. Figure 73: Mouse model of breast cancer - tumor volume. Figure 74: Mouse model of lung cancer - tumour volume. Figure 75: Mouse model of liver cancer - liver weight. Figure 76: Attachment of MRX004 and B. breve type strains to human cells. Figure 77: Exopolysaccharide production assay. Figure 78: Bound and released exopolysaccharide production by MRX004. Figure 79: Attachment of MRX004 to Caco-2 cells. Figure 80: Rapid ID 32 A profile of MRX004 alone (A) and in comparison with B. breve type strains (B). White = negative reaction (no colour change), Downwards cross-hatched = intermediate positive reaction (weak colour change) and Black = positive reaction (strong appropriate colour change). Figure 81: API ®< 50 CH analysis of MRX004. Upward cross-hatched = negative reaction (no colour change), Downward cross-hatch = intermediate positive reaction (weak colour change), Black = positive reaction (strong appropriate colour change) and White = doubtful reaction (unexpected colour change). DISCLOSURE OF THE INVENTION Bacterial strains

[0022] The compositions of the invention may comprise the bacterium deposited under accession number NCIMB 42380. The examples demonstrate that such bacteria are useful for treating or preventing diseases and conditions mediated by IL-17 or the Th17 pathway. The examples also demonstrate that such bacteria are useful for treating or preventing cancer. The preferred bacterial strain is the bacterium deposited under accession number NCIMB 42380.

[0023] The bacterium deposited under accession number NCIMB 42380 was tested in the Examples and is also referred to herein as strain 751 or MRX004. A partial 16S rRNA sequence for the 751 strain that was tested is provided in SEQ ID NO:1. Strain 751 was deposited with the international depositary authority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland) by GT Biologics Ltd. (Life Sciences Innovation Building, Aberdeen, AB25 2ZS, Scotland) on 12th March 2015 and was assigned accession number NCIMB 42380. GT Biologics Ltd. subsequently changed its name to 4D Pharma Research Limited.

[0024] A genome sequence for strain 751 is provided in SEQ ID NO:2 in the sequence listing published with PCT / GB2016 / 051776. References to SEQ ID NO:2 herein throughout, including in the description and the claims, refer to SEQ ID NO:2 of PCT / GB2016 / 051776.

[0025] Bacterial strains that are biotypes of the bacterium deposited under accession number NCIMB 42380 are also expected to be effective for treating or preventing diseases and conditions mediated by IL-17 or the Th17 pathway. Bacterial strains that are biotypes of the bacterium deposited under accession number NCIMB 42380 are also expected to be effective for treating or preventing cancer. A biotype is a closely related strain that has the same or very similar physiological and biochemical characteristics.

[0026] In certain embodiments, the bacterial strain for use in the invention has a 16s rRNA sequence that is at least 99%, 99.5% or 99.9% identical to SEQ ID NO:1. Preferably, the bacterial strain for use in the invention has the 16s rRNA sequence represented by SEQ ID NO:1.

[0027] Alternatively, strains that are biotypes of the bacterium deposited under accession number NCIMB 42380 and that are suitable for use in the invention may be identified by sequencing other nucleotide sequences for the bacterium deposited under accession number NCIMB 42380. For example substantially the whole genome may be sequenced and a biotype strain for use in the invention may have at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequence identity across at least 80% of its whole genome (e.g. across at least 85%, 90%, 95% or 99%, or across its whole genome).

[0028] The bacterial strain for use in the invention has a genome with 90% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776. The bacterial strain for use in the invention has a genome with at least 90% sequence identity (e.g. at least 92%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity) to SEQ ID NO:2 of PCT / GB2016 / 051776 across at least 60% (e.g. at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99% or 100%) of SEQ ID NO:2 of PCT / GB2016 / 051776. For example, the bacterial strain for use in the invention may have a genome with at least 90% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 70% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 90% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 80% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 90% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 90% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 90% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 100% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 95% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 70% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 95% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 80% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 95% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 90% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 95% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 100% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 98% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 70% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 98% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 80% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 98% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 90% of SEQ ID NO:2 of PCT / GB2016 / 051776, or at least 98% sequence identity to SEQ ID NO:2 of PCT / GB2016 / 051776 across 100% of SEQ ID NO:2 of PCT / GB2016 / 051776.

[0029] Alternatively, strains that are biotypes of the bacterium deposited under accession number NCIMB 42380 and that are suitable for use in the invention may be identified by using the accession number NCIMB 42380 deposit and restriction fragment analysis and / or PCR analysis, for example by using fluorescent amplified fragment length polymorphism (FAFLP) and repetitive DNA element (rep)-PCR fingerprinting, or protein profiling, or partial 16S or 23s rDNA sequencing. In preferred embodiments, such techniques may be used to identify strains of the same species as the bacterium deposited under accession number NCIMB 42380.

[0030] In certain embodiments, strains that are biotypes of the bacterium deposited under accession number NCIMB 42380 and that are suitable for use in the invention are strains that provide the same pattern as the bacterium deposited under accession number NCIMB 42380 when analysed by amplified ribosomal DNA restriction analysis (ARDRA), for example when using Sau3AI restriction enzyme (for exemplary methods and guidance see, for example,

[17] ). Alternatively, biotype strains are identified as strains that have the same carbohydrate fermentation patterns as the bacterium deposited under accession number NCIMB 42380.

[0031] Bacterial strains that are biotypes of the bacterium deposited under accession number NCIMB 42380 and that are useful in the compositions and methods of the invention may be identified using any appropriate method or strategy, including the assays described in the examples. For instance, biotypes for use in the invention may be identified by culturing in anaerobic YCFA and / or administering the bacteria to the type II collagen-induced arthritis mouse model and then assessing cytokine levels. In particular, bacterial strains that have similar growth patterns, metabolic type and / or surface antigens to the bacterium deposited under accession number NCIMB 42380 may be useful in the invention. A biotype strain will have comparable immune modulatory activity to the NCIMB 42380 strain. In particular, a biotype strain will elicit comparable effects on the asthma, arthritis, multiple sclerosis and cancer disease models and comparable effects on cytokine levels to the effects shown in the Examples, which may be identified by using the culturing and administration protocols described in the Examples.

[0032] A particularly preferred strain of the invention is the bacterium deposited under accession number NCIMB 42380. This is the exemplary 751 strain tested in the examples and shown to be effective for treating disease.

[0033] A derivative of the bacterium deposited under accession number NCIMB 42380 may be a daughter strain (progeny) or a strain cultured (subcloned) from the original. A derivative strain will have comparable immune modulatory activity to the NCIMB 42380 strain. In particular, a derivative strain will elicit comparable effects on the asthma, arthritis, multiple sclerosis and cancer disease models and same comparable effects on cytokine levels to the effects shown in the Examples, which may be identified by using the culturing and administration protocols described in the Examples. A derivative of the NCIMB 42380 strain will generally be a biotype of the NCIMB 42380 strain.

[0034] References to cells of the bacterium deposited under accession number NCIMB 42380 encompass any cells that have the same safety and therapeutic efficacy characteristics as the strain deposited under accession number NCIMB 42380, and such cells are encompassed by the invention.

[0035] In preferred embodiments, the bacterial strains in the compositions of the invention are viable and capable of partially or totally colonising the intestine.

[0036] In certain embodiments, the bacterial strain for use in the invention has low adherence to human intestinal epithelial cells, in particular Caco-2 cells. In a preferred embodiment, the bacterial strain for use in the invention has low adherence to human intestinal epithelial cells, in particular Caco-2 cells, in YCFA compared to Bifidobacteria, in particular B. breve. In certain embodiments, the bacterial strain for use in the invention exhibits adherence of less than 1% of total culture, such as preferably less than 0.5% or less than 0.3%, when tested under the conditions described in Example 12.

[0037] In certain embodiments, the bacterial strain for use in the invention produces exopolysaccharides, for example wherein the exopolysaccharides are bound to the extracellular surface of the bacterial strain. In certain embodiments, the production of the bound exopolysaccharides increases the adhesion of the bacterial strain for use in the invention to mucus or to the surface of epithelial cells, for example human intestinal epithelial cells. In a preferred embodiment, the bacterial strain for use in the invention produces more bound surface exopolysaccharides compared to Bifidobacteria, in particular B. breve.

[0038] In a preferred embodiment, the bacterial strain for use in the invention both has low adherence to human intestinal epithelial cells, in particular Caco-2 cells, in YCFA compared to Bifidobacteria, in particular B. breve (such as adherence of less than 1% of total culture, such as preferably less than 0.5% or less than 0.3%, when tested under the conditions described in Example 12), and produces more bound surface exopolysaccharides compared to Bifidobacteria, in particular B. breve.

[0039] In certain preferred embodiments, the bacterial strain for use in the invention is able to ferment the polysaccharide raffinose, for example when cultured in an appropriate suspension medium (such as API suspension medium) at 37°C for 4 hours.

[0040] In certain embodiments, the bacterial strain for use in the invention has reduced ability to ferment α-glucosidase and / or β-glucosidase compared to Bifidobacteria, in particular B. breve, for example when cultured in an appropriate suspension medium (such as API suspension medium) at 37°C for 4 hours.

[0041] In certain embodiments, the bacterial strain for use in the invention comprises one or more of the genes listed in Table 1, such as 5, 10, 20, 50 or all of the genes in Table 1. In certain embodiments, the bacterial strain for use in the invention comprises one or more of the genes listed in Table 1 that are highlighted with single underlining, such as Transmembrane component BL0694 of energizing module of predicted ECF transporter and / or Duplicated ATPase component BL0693 of energizing module of predicted ECF transporter. In certain embodiments, the bacterial strain for use in the invention comprises one or more of the genes listed in Table 1 that are highlighted with double underlining and in bold, such as 1, 2, 3, 4 or 5 genes selected from: maltodextrin glucosidase (EC 3.2.1.20), putative galactosidase, cellulose synthase (UDP-forming) (EC 2.4.1.12), chitinase (EC 3.2.1.14) and sensory box / GGDEF family protein. In certain embodiments, the bacterial strain for use in the invention comprises one or more of the genes listed in Table 1 that are highlighted with italics, such as 1, 2, 3, 4, 5, 6, 7, 8 or 9 genes selected from: omega-3 polyunsaturated fatty acid synthase subunit PfaA, Type I polyketide synthase, putative glycosyl hydrolase of unknown function (DUF1680), ATPase component BioM of energizing module of biotin ECF transporter, Cation-transporting ATPase E1-E2 family, Ribose ABC transport system permease protein RbsC (TC 3.A.1.2.1), Ribose ABC transport system ATP-binding protein RbsA (TC 3.A.1.2.1), 3'-to-5' oligoribonuclease (orn), membrane protein related to Actinobacillus protein (1944168).

[0042] In preferred embodiments, the bacterial strain for use in the invention comprises one or more (such as 5, 10, 15, 20, 25, 30, 40, 45, 50 or all) genes selected from: 2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylic-acid synthase (EC 2.2.1.9); 3'-to-5' oligoribonuclease (orn); Alpha-galactosidase (EC 3.2.1.22); ATPase component of general energizing module of ECF transporters; ATPase component STY3233 of energizing module of queuosine-regulated ECF transporter; ATP-dependent DNA helicase recG (EC 3.6.1.-); Beta-glucosidase (EC 3.2.1.21); Cellulose synthase (UDP-forming) (EC 2.4.1.12); Chitinase (EC 3.2.1.14); COG1309: Transcriptional regulator; D-alanyl-D-alanine carboxypeptidase (EC 3.4.16.4); Duplicated ATPase component BL0693 of energizing module of predicted ECF transporter; Fructokinase (EC 2.7.1.4); Glucose / mannose:H+ symporter GlcP; Glycosyltransferase (EC 2.4.1.-); GMP synthase [glutamine-hydrolyzing] (EC 6.3.5.2); Hypothetical sugar kinase in cluster with indigoidine synthase indA , PfkB family of kinases; Inosine-uridine preferring nucleoside hydrolase (EC 3.2.2.1); LSU ribosomal protein L31p @ LSU ribosomal protein L31p, zinc-independent; LSU ribosomal protein L33p @ LSU ribosomal protein L33p, zinc-independent; Maltodextrin glucosidase (EC 3.2.1.20); Membrane protein, related to Actinobacillus protein (1944168); Membrane-bound lytic murein transglycosylase D precursor (EC 3.2.1.-); Methyltransferase (EC 2.1.1.-); NADH-dependent butanol dehydrogenase A (EC 1.1.1.-); Phosphoglycolate phosphatase (EC 3.1.3.18); Phosphoribosylanthranilate isomerase (EC 5.3.1.24); Putative glycosyl hydrolase of unknown function (DUF1680); Rhamnose-containing polysacharide translocation permease; Ribokinase (EC 2.7.1.15); Ribose ABC transport system, ATP-binding protein RbsA (TC 3.A.1.2.1); Ribose ABC transport system, ATP-binding protein RbsA (TC 3.A.1.2.1); Ribose ABC transport system, high affinity permease RbsD (TC 3.A.1.2.1); Ribose ABC transport system, periplasmic ribose-binding protein RbsB (TC 3.A.1.2.1); Ribose ABC transport system, permease protein RbsC (TC 3.A.1.2.1); Ribose ABC transport system, permease protein RbsC (TC 3.A.1.2.1); Sorbitol dehydrogenase (EC 1.1.1.14); SSU ribosomal protein S14p (S29e) @ SSU ribosomal protein S14p (S29e), zinc-independent; Substrate-specific component STY3230 of queuosine-regulated ECF transporter; Sucrose-6-phosphate hydrolase (EC 3.2.1.B3); Teichoic acid export ATP-binding protein TagH (EC 3.6.3.40); Transmembrane component BL0694 of energizing module of predicted ECF transporter; Transmembrane component STY3231 of energizing module of queuosine-regulated ECF transporter; Two-component response regulator colocalized with HrtAB transporter; Type I restriction-modification system, DNA-methyltransferase subunit M (EC 2.1.1.72); Type I restriction-modification system, restriction subunit R (EC 3.1.21.3); Type I restriction-modification system, specificity subunit S (EC 3.1.21.3); Type I restriction-modification system, specificity subunit S (EC 3.1.21.3); Type I restriction-modification system, specificity subunit S (EC 3.1.21.3); Xylitol dehydrogenase (EC 1.1.1.9); and Xylose ABC transporter, periplasmic xylose-binding protein XylF. In preferred embodiments, the bacterial strain for use in the invention comprises one or more (such as 5, 10, 15, 20, 25, 30, 35 or all) genes that are listed in the preceding sentence and that are not highlighted in Table 1.Therapeutic uses

[0043] As demonstrated in the examples, the bacterial compositions of the invention are effective for reducing the Th17 inflammatory response. In particular, treatment with compositions of the invention achieves a reduction in IL-17A levels and other Th17 pathway cytokines, and clinical improvements in animal models of conditions mediated by IL-17 and the Th17 pathway.

[0044] Th17 cells are a subset of T helper cells that produce, for example, IL-17A, IL17-F, IL-21 and IL-22. Th17 cell differentiation and IL-17 expression may be driven by IL-23. These cytokines and others form important parts of the Th17 pathway, which is a well-established inflammatory signalling pathway that contributes to and underlies a number of inflammatory and autoimmune diseases (as described in, for example, [18-23]). Diseases wherein the Th17 pathway is activated are Th17 pathway-mediated diseases. Th17 pathway-mediated diseases can be ameliorated or alleviated by repressing the Th17 pathway, which may be through a reduction in the differentiation of Th17 cells or a reduction in their activity or a reduction in the level of Th17 pathway cytokines. Diseases mediated by the Th17 pathway may be characterised by increased levels of cytokines produced by Th17 cells, such as IL-17A, IL-17F, IL-21, IL-22, IL-26, IL-9 (reviewed in

[24] ). Diseases mediated by the Th17 pathway may be characterised by increased expression of Th-17-related genes, such as Stat3 or IL-23R. Diseases mediated by the Th17 pathway may be associated with increased levels of Th17 cells.

[0045] IL-17 is a pro-inflammatory cytokine that contributes to the pathogenesis of several inflammatory and autoimmune diseases and conditions. IL-17 as used herein may refer to any member of the IL-17 family, including IL-17A, IL-17B, IL-17C, IL-17D, IL-17E, and IL-17F. IL-17-mediated diseases and conditions are characterised by high expression of IL-17 and / or the accumulation or presence of IL-17-positive cells in a tissue affected by the disease or condition. Similarly, IL-17-mediated diseases and conditions are diseases and conditions that are exacerbated by high IL-17 levels or an increase in IL-17 levels, and that are alleviated by low IL-17 levels or a reduction in IL-17 levels. The IL-17 inflammatory response may be local or systemic.

[0046] Examples of diseases and conditions that may be mediated by IL-17 or the Th17 pathway include multiple sclerosis; arthritis, such as rheumatoid arthritis, osteoarthritis, psoriatic arthritis, or juvenile idiopathic arthritis; neuromyelitis optica (Devic's disease); ankylosing spondylitis; spondyloarthritis; psoriasis; systemic lupus erythematosus; inflammatory bowel disease, such as Crohn's disease or ulcerative colitis; celiac disease; asthma, such as allergic asthma or neutrophilic asthma; chronic obstructive pulmonary disease (COPD); cancer, such as breast cancer, colon cancer, lung cancer or ovarian cancer; uveitis; scleritis; vasculitis; Behcet's disease; atherosclerosis; atopic dermatitis; emphysema; periodontitis; allergic rhinitis; and allograft rejection.

[0047] IL-17 and the Th17 pathway are often associated with chronic inflammatory and autoimmune diseases, so the compositions of the invention may be particularly useful for treating or preventing chronic diseases or conditions. In certain embodiments, the compositions are for use in patients with chronic disease. In certain embodiments, the compositions are for use in preventing the development of chronic disease.

[0048] In certain embodiments, treatment with compositions of the invention provides a reduction or prevents an elevation in IL-17 levels, in particular IL-17A levels. In certain embodiments, treatment with compositions of the invention provides a reduction or prevents an elevation in IFN-γ, IL,-1β, RANTES, MIP-1α, IL-8 or IL-6 levels. Such reduction or prevention of elevated levels of these cytokines may be useful for treating or preventing inflammatory and autoimmune diseases and conditions, in particular those mediated by IL-17 or the Th17 pathway.

[0049] In certain embodiments, treatment with the compositions of the invention provides a block of the attachment to or invasion of human cells, for example human epithelial cells by pathogenic cells, for example E. coli and / or S. enteritidis.

[0050] In certain embodiments, treatment with the compositions of the invention reduces or prevents the binding of pathogenic cells, for example E. coli and / or S. enteritidis, to the human epithelial cells, for example human intestinal epithelial cells.

[0051] In certain embodiments, the production and release of exopolysaccharides by the bacterial strains of the compositions of the invention may have protective effects against pathogenic species, for example E. coli and / or S. enteritidis. In certain embodiments, the production and release of exopolysaccharides by the bacterial strains of the compositions of the invention may mediate the effect of the bacteria on the IL-17 or Th17 pathway and may influence the host immune response. In certain embodiments, the compositions of the invention are for use in producing exopolysaccharides in the treatment of inflammatory and autoimmune diseases, and in particular diseases or conditions mediated by IL-17.

[0052] In certain embodiments, the low adherence to human intestinal epithelial cells, in particular Caco-2 cells, of the bacterial strains of the compositions of the invention may increase the beneficial effect of the compositions of the invention on the IL-17 or the Th17 pathway and on diseases mediated by IL-17 or the Thl7 pathway.

[0053] In certain embodiments, treatment with compositions of the invention provides increased fermentation of raffinose in the intestine. The examples demonstrate that the bacterial strains of the compositions of the invention ferment the polysaccharide raffinose, and raffinose fermentation may confer effects on the host such as increased caecal butyrate and increased gastrointestinal proliferation. In certain embodiments, the compositions of the invention are for use in increasing fermentation of raffinose in the intestine in the treatment of inflammatory and autoimmune diseases, and in particular diseases or conditions mediated by IL-17.Asthma

[0054] In preferred embodiments, the compositions of the invention are for use in treating or preventing allergic or eosinophilic asthma. The examples demonstrate that the compositions of the invention achieve a reduction in the recruitment of neutrophils and / or eosinophils into the airways following sensitisation and challenge with house dust mite extract and so they may be useful in the treatment or prevention of asthma. Asthma is a chronic disease characterised by inflammation and restriction of the airways. The inflammation in asthma may be mediated by eosinophils and / or neutrophils.

[0055] In certain embodiments, the asthma is eosinophilic or allergic asthma. Eosinophilic and allergic asthma are characterised by increased numbers of eosinophils in peripheral blood and in airway secretions and is associated pathologically with thickening of the basement membrane zone and pharmacologically by corticosteroid responsiveness

[25] . Compositions that reduce or inhibit eosinophil recruitment or activation may be useful for treating or preventing eosinophilic and allergic asthma.

[0056] Eosinophilic and neutrophilic asthma are not mutually exclusive conditions and treatments that help address either the eosinophil and neutrophil responses may be useful for treating asthma in general.

[0057] In certain embodiments, the compositions of the invention are for use in methods of reducing an eosinophilic inflammatory response in the treatment or prevention of eosinophilic asthma. As noted above, high levels of eosinophils in asthma is associated pathologically with thickening of the basement membrane zone, so reducing eosinophilic inflammatory response in the treatment or prevention of asthma may be able to specifically address this feature of the disease. Also, elevated neutrophils, either in combination with elevated eosinophils or in their absence, is associated with severe asthma and chronic airway narrowing. Therefore, reducing the neutrophilic inflammatory response may be particularly useful for addressing severe asthma.

[0058] In certain embodiments, the compositions reduce peribronchiolar infiltration in allergic asthma, or are for use in reducing peribronchiolar infiltration in the treatment of allergic asthma.

[0059] In certain embodiments, treatment with compositions of the invention provides a reduction or prevents an elevation in IL-1β, IFNγ, RANTES, MIP-1α or IL-8 levels.

[0060] In certain embodiments, the compositions of the invention are for use in a method of treating eosinophilic asthma that results in a reduction of the eosinophilic inflammatory response. In certain embodiments, the patient to be treated has, or has previously been identified as having, elevated neutrophil or eosinophil levels, for example as identified through blood sampling or sputum analysis.

[0061] The compositions of the invention may be useful for preventing the development of allergic or eosinophilic asthma in a new-born when administered to the new-born, or to a pregnant woman. The compositions may be useful for preventing the development of allergic or eosinophilic asthma in children. The compositions of the invention may be useful for treating or preventing adult-onset allergic or eosinophilic asthma. The compositions of the invention may be useful for managing or alleviating allergic or eosinophilic asthma. The compositions of the invention may be particularly useful for reducing symptoms associated with allergic or eosinophilic asthma that is aggravated by allergens, such as house dust mites.

[0062] Treatment or prevention of asthma may refer to, for example, an alleviation of the severity of symptoms or a reduction in the frequency of exacerbations or the range of triggers that are a problem for the patient.

[0063] In certain embodiments, treatment with compositions of the invention provides a reduction in concentrations of phenylalanine and / or histidine, for example in the intestines or in the plasma. The examples demonstrate that the bacterial strains of the compositions of the invention tested positive for fermentation of amino acids, including phenylalanine and histidine, and increased plasma concentrations of phenylalanine and histidine have been reported to be associated with adverse effects in asthma. In certain embodiments, the compositions of the invention are for use in reducing airway hyper-responsiveness associated with asthma.

[0064] In certain embodiments, treatment with compositions of the invention provides a reduction in concentrations of galactose and / or fructose, for example in the intestines. The examples demonstrate that the bacterial strains of the compositions of the invention ferment carbohydrate substrates including galactose and fructose, and galactose α-1,3- galactose derived from meat sources is a known allergen and causative agent of anaphylaxis, and intake levels of dietary fructose are correlated with increased asthma severity.Modes of administration

[0065] Preferably, the compositions of the invention are to be administered to the gastrointestinal tract in order to enable delivery to and / or partial or total colonisation of the intestine with the bacterial strain of the invention. Generally, the compositions of the invention are administered orally, but they may be administered rectally, intranasally, or via buccal or sublingual routes.

[0066] In certain embodiments, the compositions of the invention may be administered as a foam, as a spray or a gel.

[0067] In certain embodiments, the compositions of the invention may be administered as a suppository, such as a rectal suppository, for example in the form of a theobroma oil (cocoa butter), synthetic hard fat (e.g. suppocire, witepsol), glycero-gelatin, polyethylene glycol, or soap glycerin composition.

[0068] In certain embodiments, the composition of the invention is administered to the gastrointestinal tract via a tube, such as a nasogastric tube, orogastric tube, gastric tube, jejunostomy tube (J tube), percutaneous endoscopic gastrostomy (PEG), or a port, such as a chest wall port that provides access to the stomach, jejunum and other suitable access ports.

[0069] The compositions of the invention may be administered once, or they may be administered sequentially as part of a treatment regimen. In certain embodiments, the compositions of the invention are to be administered daily.

[0070] In certain embodiments of the invention, treatment according to the invention is accompanied by assessment of the patient's gut microbiota. Treatment may be repeated if delivery of and / or partial or total colonisation with the strain of the invention is not achieved such that efficacy is not observed, or treatment may be ceased if delivery and / or partial or total colonisation is successful and efficacy is observed.

[0071] In certain embodiments, the composition of the invention may be administered to a pregnant animal, for example a mammal such as a human in order to prevent an inflammatory or autoimmune disease developing in her child in utero and / or after it is born.

[0072] The compositions of the invention may be administered to a patient that has been diagnosed with a disease or condition mediated by IL-17 or the Th17 pathway, or that has been identified as being at risk of a disease or condition mediated by IL-17 or the Th17 pathway. The compositions may also be administered as a prophylactic measure to prevent the development of diseases or conditions mediated by IL-17 or the Th17 pathway in a healthy patient.

[0073] The compositions of the invention may be administered to a patient that has been identified as having an abnormal gut microbiota. For example, the patient may have reduced or absent colonisation by the bacterium deposited under accession number NCIMB 42380.

[0074] The compositions of the invention may be administered as a food product, such as a nutritional supplement.

[0075] Generally, the compositions of the invention are for the treatment of humans, although they may be used to treat animals including monogastric mammals such as poultry, pigs, cats, dogs, horses or rabbits. The compositions of the invention may be useful for enhancing the growth and performance of animals. If administered to animals, oral gavage may be used.Compositions

[0076] Generally, the composition of the invention comprises bacteria. In preferred embodiments of the invention, the composition is formulated in freeze-dried form. For example, the composition of the invention may comprise granules or gelatin capsules, for example hard gelatin capsules, comprising a bacterial strain of the invention.

[0077] Preferably, the composition of the invention comprises lyophilised bacteria. Lyophilisation of bacteria is a well-established procedure and relevant guidance is available in, for example, references [51-53].

[0078] Alternatively, the composition of the invention may comprise a live, active bacterial culture.

[0079] In preferred embodiments, the composition of the invention is encapsulated to enable delivery of the bacterial strain to the intestine. Encapsulation protects the composition from degradation until delivery at the target location through, for example, rupturing with chemical or physical stimuli such as pressure, enzymatic activity, or physical disintegration, which may be triggered by changes in pH. Any appropriate encapsulation method may be used. Exemplary encapsulation techniques include entrapment within a porous matrix, attachment or adsorption on solid carrier surfaces, self-aggregation by flocculation or with cross-linking agents, and mechanical containment behind a microporous membrane or a microcapsule. Guidance on encapsulation that may be useful for preparing compositions of the invention is available in, for example, references

[54] and

[55] .

[0080] The composition may be administered orally and may be in the form of a tablet, capsule or powder. Encapsulated products are preferred because the bacterium deposited under accession number NCIMB 42380 may be an anaerobe. Other ingredients (such as vitamin C, for example), may be included as oxygen scavengers and prebiotic substrates to improve the delivery and / or partial or total colonisation and survival in vivo. Alternatively, the probiotic composition of the invention may be administered orally as a food or nutritional product, such as milk or whey based fermented dairy product, or as a pharmaceutical product.

[0081] The composition may be formulated as a probiotic.

[0082] A composition of the invention includes a therapeutically effective amount of a bacterial strain of the invention. A therapeutically effective amount of a bacterial strain is sufficient to exert a beneficial effect upon a patient. A therapeutically effective amount of a bacterial strain may be sufficient to result in delivery to and / or partial or total colonisation of the patient's intestine.

[0083] A suitable daily dose of the bacteria, for example for an adult human, may be from about 1 × 10 3< to about 1 × 10 11< colony forming units (CFU); for example, from about 1 × 10 7< to about 1 × 10 10< CFU; in another example from about 1 × 10 6< to about 1 × 10 10< CFU.

[0084] In certain embodiments, the composition contains the bacterial strain in an amount of from about 1 × 10 6< to about 1 × 10 11< CFU / g, respect to the weight of the composition; for example, from about 1 × 10 8< to about 1 × 10 10< CFU / g. The dose may be, for example, 1 g, 3g, 5g, and 10g.

[0085] Typically, a probiotic, such as the composition of the invention, is optionally combined with at least one suitable prebiotic compound. A prebiotic compound is usually a non-digestible carbohydrate such as an oligo- or polysaccharide, or a sugar alcohol, which is not degraded or absorbed in the upper digestive tract. Known prebiotics include commercial products such as inulin and transgalacto-oligosaccharides.

[0086] In certain embodiments, the probiotic composition of the present invention includes a prebiotic compound in an amount of from about 1 to about 30% by weight, respect to the total weight composition, (e.g. from 5 to 20% by weight). Carbohydrates may be selected from the group consisting of: fructo- oligosaccharides (or FOS), short-chain fructo-oligosaccharides, inulin, isomaltoligosaccharides, pectins, xylo-oligosaccharides (or XOS), chitosan-oligosaccharides (or COS), beta-glucans, arable gum modified and resistant starches, polydextrose, D-tagatose, acacia fibers, carob, oats, and citrus fibers. In one aspect, the prebiotics are the short-chain fructo-oligosaccharides (for simplicity shown herein below as FOSs-c.c); said FOSs-c.c. are not digestible carbohydrates, generally obtained by the conversion of the beet sugar and including a saccharose molecule to which three glucose molecules are bonded.

[0087] The compositions of the invention may comprise pharmaceutically acceptable excipients or carriers. Examples of such suitable excipients may be found in the reference

[56] . Acceptable carriers or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in reference

[57] . Examples of suitable carriers include lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol, sorbitol and the like. Examples of suitable diluents include ethanol, glycerol and water. The choice of pharmaceutical carrier, excipient or diluent can be selected with regard to the intended route of administration and standard pharmaceutical practice. The pharmaceutical compositions may comprise as, or in addition to, the carrier, excipient or diluent any suitable binder(s), lubricant(s), suspending agent(s), coating agent(s), solubilising agent(s). Examples of suitable binders include starch, gelatin, natural sugars such as glucose, anhydrous lactose, free-flow lactose, beta-lactose, corn sweeteners, natural and synthetic gums, such as acacia, tragacanth or sodium alginate, carboxymethyl cellulose and polyethylene glycol. Examples of suitable lubricants include sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride and the like. Preservatives, stabilizers, dyes and even flavouring agents may be provided in the pharmaceutical composition. Examples of preservatives include sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid. Antioxidants and suspending agents may be also used.

[0088] The compositions of the invention may be formulated as a food product. For example, a food product may provide nutritional benefit in addition to the therapeutic effect of the invention, such as in a nutritional supplement. Similarly, a food product may be formulated to enhance the taste of the composition of the invention or to make the composition more attractive to consume by being more similar to a common food item, rather than to a pharmaceutical composition. In certain embodiments, the composition of the invention is formulated as a milk-based product. The term "milk-based product" means any liquid or semi-solid milk- or whey- based product having a varying fat content. The milk-based product can be, e.g., cow's milk, goat's milk, sheep's milk, skimmed milk, whole milk, milk recombined from powdered milk and whey without any processing, or a processed product, such as yoghurt, curdled milk, curd, sour milk, sour whole milk, butter milk and other sour milk products. Another important group includes milk beverages, such as whey beverages, fermented milks, condensed milks, infant or baby milks; flavoured milks, ice cream; milk-containing food such as sweets.

[0089] In certain embodiments, the compositions of the invention contain a single bacterial strain or species and do not contain any other bacterial strains or species. Such compositions may comprise only de minimis or biologically irrelevant amounts of other bacterial strains or species. Such compositions may be a culture that is substantially free from other species of organism.

[0090] The compositions for use in accordance with the invention may or may not require marketing approval.

[0091] In some cases, the lyophilised bacterial strain is reconstituted prior to administration. In some cases, the reconstitution is by use of a diluent described herein.

[0092] The compositions of the invention can comprise pharmaceutically acceptable excipients, diluents or carriers.

[0093] In certain embodiments, the invention provides the above pharmaceutical composition, wherein the amount of the bacterial strain is from about 1 × 10 3< to about 1 × 10 11< colony forming units per gram with respect to a weight of the composition.

[0094] In certain embodiments, the invention provides the above pharmaceutical composition, wherein the composition is administered at a dose of 1 g, 3 g, 5 g or 10 g.

[0095] In certain embodiments, the invention provides the above pharmaceutical composition, wherein the composition is administered by a method selected from the group consisting of oral, rectal, subcutaneous, nasal, buccal, and sublingual.

[0096] In certain embodiments, the invention provides the above pharmaceutical composition, comprising a carrier selected from the group consisting of lactose, starch, glucose, methyl cellulose, magnesium stearate, mannitol and sorbitol.

[0097] In certain embodiments, the invention provides the above pharmaceutical composition, comprising a diluent selected from the group consisting of ethanol, glycerol and water.

[0098] In certain embodiments, the invention provides the above pharmaceutical composition, comprising an excipient selected from the group consisting of starch, gelatin, glucose, anhydrous lactose, free-flow lactose, beta-lactose, corn sweetener, acacia, tragacanth, sodium alginate, carboxymethyl cellulose, polyethylene glycol, sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate and sodium chloride.

[0099] In certain embodiments, the invention provides the above pharmaceutical composition, further comprising at least one of a preservative, an antioxidant and a stabilizer.

[0100] In certain embodiments, the invention provides the above pharmaceutical composition, comprising a preservative selected from the group consisting of sodium benzoate, sorbic acid and esters of p-hydroxybenzoic acid.

[0101] In certain embodiments, the invention provides the above pharmaceutical composition, wherein said bacterial strain is lyophilised.

[0102] In certain embodiments, the invention provides the above pharmaceutical composition, wherein when the composition is stored in a sealed container at about 4°C or about 25°C and the container is placed in an atmosphere having 50% relative humidity, at least 80% of the bacterial strain as measured in colony forming units, remains after a period of at least about: 1 month, 3 months, 6 months, 1 year, 1.5 years, 2 years, 2.5 years or 3 years.Culturing methods

[0103] The bacterial strains for use in the present invention can be cultured using standard microbiology techniques as detailed in, for example, references [58-60].

[0104] The solid or liquid medium used for culture may be YCFA agar or YCFA medium. YCFA medium may include (per 100ml, approximate values): Casitone (1.0 g), yeast extract (0.25 g), NaHCO 3 (0.4 g), cysteine (0.1 g), K 2 HPO 4 (0.045 g), KH 2 PO 4 (0.045 g), NaCl (0.09 g), (NH 4 ) 2 SO 4 (0.09 g), MgSO 4 ·7H 2 O (0.009 g), CaCl 2 (0.009 g), resazurin (0.1 mg), hemin (1 mg), biotin (1 µg), cobalamin (1 µg), p-aminobenzoic acid (3 µg), folic acid (5 µg), and pyridoxamine (15 µg).General

[0105] The practice of the present invention will employ, unless otherwise indicated, conventional methods of chemistry, biochemistry, molecular biology, immunology and pharmacology, within the skill of the art. Such techniques are explained fully in the literature. See, e.g., references

[61] and [62-68], etc.

[0106] The term "comprising" encompasses "including" as well as "consisting" e.g. a composition "comprising" X may consist exclusively of X or may include something additional e.g. X + Y.

[0107] The term "about" in relation to a numerical value x is optional and means, for example, x+10%.

[0108] The word "substantially" does not exclude "completely" e.g. a composition which is "substantially free" from Y may be completely free from Y. Where necessary, the word "substantially" may be omitted from the definition of the invention.

[0109] References to a percentage sequence identity between two nucleotide sequences means that, when aligned, that percentage of nucleotides are the same in comparing the two sequences. This alignment and the percent homology or sequence identity can be determined using software programs known in the art, for example those described in section 7.7.18 of ref.

[69] . A preferred alignment is determined by the Smith-Waterman homology search algorithm using an affine gap search with a gap open penalty of 12 and a gap extension penalty of 2, BLOSUM matrix of 62. The Smith-Waterman homology search algorithm is disclosed in ref.

[70] .

[0110] Unless specifically stated, a process or method comprising numerous steps may comprise additional steps at the beginning or end of the method, or may comprise additional intervening steps. Also, steps may be combined, omitted or performed in an alternative order, if appropriate.

[0111] Various embodiments of the invention are described herein. It will be appreciated that the features specified in each embodiment may be combined with other specified features, to provide further embodiments. In particular, embodiments highlighted herein as being suitable, typical or preferred may be combined with each other (except when they are mutually exclusive).EXAMPLES Example 1 - Efficacy of bacterial inocula in a mouse model of house dust mite-induced asthma Summary

[0112] Mice were administered with compositions comprising bacterial strains according to the invention and were subsequently challenged with house dust mite (HDM) extract to elicit an allergic inflammatory response. The inflammatory response to HDM includes eosinophilic and neutrophilic components, is mediated by IL-17 and the Th17 pathway, and is a model for asthma. The magnitude and characteristics of the inflammatory response exhibited by mice treated with compositions of the invention were compared to control groups. The compositions of the invention were found to alleviate the inflammatory response, and to reduce recruitment of eosinophils and neutrophils, indicating that they may be useful for treating IL-17-mediated conditions such as eosinophilia, neutrophilia and asthma.Strain

[0113] 751: bacterium deposited under accession number NCIMB 42380Study designGroups:

[0114] 1. Negative control group. Treatment with vehicle control (per oral). 4. Treatment with therapeutic bacteria inoculum strain 751 (per oral). 7. Positive control group. Treatment with Dexamethasone (i.p.). 8. Untreated Control Group. Number of mice per group = 5 Day -14 to day 13: Daily administration of vehicle control per oral (Group 1). Day -14 to day 13: Daily administration of therapeutic bacteria inoculum per oral (Group 2-6). Day 0, 2, 4, 7, 9, 11 Administration of 15ug HDM (house dust mite extract - Catalogue number: XPB70D3A25, Lot number: 231897, Greer Laboratories, Lenoir, NC, USA) in a volume of 30ul PBS per nasal (Group 1-8). Day 0, 2, 4, 7, 9, 11 Administration of Dexamethasone (i.p., 3mg / kg, Sigma-Aldrich, Catalogue number D1159) (Group 7). Day 14 Sacrifice of all animals for analysis. Total number of mice = 40. Endpoints and analysis

[0115] On day 14 animals were sacrificed by lethal intraperitoneal injection with pentabarbitol (Streuli Pharma AG, Uznach, Cat: 1170139A) immediately followed by a bronchoalveolar lavage (BAL).

[0116] Cells were isolated from the BAL (bronchoalveolar lavage) fluid and differential cell counts performed (200 cell counts / samples).Material and Methods

[0117] Mice . Female 7 week old BALB / c mice were purchased from Charles River Laboratories and randomly allocated to cages totally 5 mice per cage (Ventilated cages sourced from Indulab AG, Gams, Switzerland Cage type: "The SealsafeTM - IVC cage. Product number 1248L). Cages were labeled with study number, group number and experimental starting date. Mice were monitored weekly and acclimatized to facility for 7 days prior to initiation of study (Study Day -14). Animals were 8 weeks old on Study Day -14. Potable water and food were available ad libitum. Cage enrichment was present. Daily care of the animals was performed according to local authorization license number 2283.1 (issued and approved by: Service de la consommation et des affaires vétérinaires du Canton de Vaud). Potable water and food were available ad libitum and refreshed once daily. Cage enrichment was present. Animal welfare regulations were observed as given by official authorities of Switzerland under ordinance 455.163 of the FVO (Federal Veterinary Office) on laboratory animal husbandry, production of genetically modified animals, and methods of animal experimentation.

[0118] Culturing of bacteria inoculum . Within a sterile workstation, a cryo-vial of bacteria was thawed by warming in gloved hand and ~0.7 ml of contents injected into a Hungate tube (Cat Number, 1020471, Glasgerätebau Ochs, Bovenden-Lenglern, Germany), containing 8 ml of anaerobic YCFA. Two tubes per strain were usually prepared. The Hungate tubes were then incubated (static) at 37°C for 16h (strain 751).

[0119] Culturing of vehicle control . A Hungate tube containing 8 ml of anaerobic YCFA was incubated (static) at 37°C for 16h.

[0120] Administration of bacteria inoculum or vehicle control . 400ul of cultured bacteria inoculum or vehicle control were administered per day per oral gavage.

[0121] Intranasal sensitization. Mice were anesthetized by i.p. injection with 9.75 mg xylasol and 48.75 mg ketasol per kg (Dr. E. Graeub AG, Bern, Switzerland) and administered with 15ug of HDM (Catalogue number: XPB70D3A25, Lot number: 231897, Greer Laboratories, Lenoir, NC, USA) in a volume of 30ul PBS per nasal.

[0122] Preparation and administration of positive control compound Dexamethasone . Dexamethasone 21-phosphate disodium salt (Sigma-Aldrich, Catalogue number D1159, Lot N° SLBD.1030V) was solved in H 2 O and administered to the animals in a dose of 3mg / kg in a volume of 200ul per oral at days indicated in study protocol above.

[0123] Terminal procedure. On day 14 animals were sacrificed by lethal i.p. injection with pentabarbitol (Streuli Pharma AG, Uznach, Cat: 117013 9A) immediately followed by bronchoalveolar lavage (BAL) in 500 ul of saline.

[0124] Measurement of cellular infiltrates into BAL. Cells were isolated from the BAL fluid and differential cell counts were performed based upon standard morphological and cytochemical criteria.

[0125] Graphs and statistical analysis . All graphs were generated with Graphpad Prism Version 6 and a one-way ANOVA was applied. Results from the statistical analysis were provided with the individual data tables. Error bars represent Standard Error of the Mean (SEM).Results and analysis

[0126] The results of the experiments are shown in Figures 1-9.

[0127] No morbidity or mortality was noted in the mice treated with the bacteria or the vehicle. The two controls, vehicle treatment (negative control) and the dexamethasone treatment (positive control) behaved as expected, with impaired eosinophilia and neutrophilia noted following dexamethasone treatment.

[0128] The most important results of this experiment are displayed in Figures 6 and 7, which report on the total number and percentage of neutrophils detected in bronchiolar lavage following challenge with HDM. Strain 751 reduced total neutrophils and the proportion of neutrophils in BAL relative to the vehicle-only control.Example 2 - Efficacy of bacterial inocula in a mouse model of severe neutrophilic asthma Summary

[0129] Mice were administered with compositions comprising bacterial strains according to the invention and were subsequently sensitised with subcutaneous administrations of house dust mite (HDM) extract and challenged with an intranasal administration of HDM in order to model the inflammatory response of severe neutrophilic asthma. The magnitude and characteristics of the inflammatory response exhibited by mice treated with compositions of the invention were compared to control groups. The compositions of the invention were found to alleviate the inflammatory response, and in particular to reduce recruitment of neutrophils, in a manner comparable to the positive control comprising administrations of anti-IL-17 antibodies. The data therefore indicate that the compositions of the invention may be useful for treating IL-17- and Th17-mediated conditions such as neutrophilia and asthma.Strain751: bacterium deposited under accession number NCIMB 42380Study designGroups:

[0130] 1. Negative control group. Treatment with vehicle control (per oral). 4. Treatment with therapeutic bacteria inoculum strain 751 (per oral). 7. Positive control group. Treatment anti-IL-17 (i.p.). 8. Untreated Control Group. 9: Healthy mice (baseline). Number of mice per group (Group 1-8) = 5 Day -14 to day 17: Daily administration of vehicle control per oral (Group 1). Day -14 to day 17: Daily administration of therapeutic bacteria inoculum per oral (Group 2-6). Day 0: Sensitization with HDM in CFA (s.c.) (Group 1-8). Day 7: Sensitization with HDM in CFA (s.c.) (Group 1-8). Day 13, 15, 17: Administration of anti IL-17 neutralizing antibody per i.p. (Group 7). Day 14, 15, 16, 17: Challenge with HDM in 30ul PBS per nasal (Group 1-8). Day 18: Sacrifice of all animals for analysis. Endpoints and analysis:

[0131] On day 14 animals were sacrificed by lethal intraperitoneal injection with pentabarbitol (Streuli Pharma AG, Uznach, Cat: 1170139A) immediately followed by a bronchoalveolar lavage (BAL). Cells were isolated from the BAL fluid and differential cell counts performed (200 cell counts / samples).Material and Methods.

[0132] Mice. Female 7 week old C57BL / 6 mice were purchased from Charles River Laboratories and randomly allocated to cages totally 5 mice per cage (Ventilated cages sourced from Indulab AG, Gams, Switzerland Cage type: "The SealsafeTM - IVC cage. Product number 1248L). Cages were labelled with study number, group number and experimental starting date. Mice were monitored weekly and acclimatized to facility for 7 days prior to initiation of study (Study Day -14). Animals were 8 weeks old on Study Day -14. Potable water and food were available ad libitum. Cage enrichment was present. Daily care of the animals was performed according to local authorization license number 2283.1 (issued and approved by: Service de la consommation et des affaires vétérinaires du Canton de Vaud). Potable water and food were available ad libitum and refreshed once daily. Cage enrichment was present. Animal welfare regulations were observed as given by official authorities of Switzerland under ordinance 455.163 of the FVO (Federal Veterinary Office) on laboratory animal husbandry, production of genetically modified animals, and methods of animal experimentation.

[0133] Culturing of bacteria inoculum . Within a sterile workstation, a cryo-vial of bacteria was thawed by warming in gloved hand and ~0.7 ml of contents injected into a Hungate tube (Cat Number, 1020471, Glasgerätebau Ochs, Bovenden-Lenglern, Germany), containing 8 ml of anaerobic YCFA. Two tubes per strain were usually prepared. The Hungate tubes were then incubated (static) at 37°C for 16h (strain 751).

[0134] Culturing of vehicle control . A Hungate tube containing 8 ml of anaerobic YCFA was incubated (static) at 37°C for 16h.

[0135] Administration of bacteria inoculum or vehicle control . 400ul of cultured bacteria inoculum or vehicle control were administered per day per oral gavage.

[0136] HDM sensitization. 50 µg of HDM (Catalogue number: XPB70D3A25, Lot number: 231897, Greer Laboratories, Lenoir, NC, USA) in PBS was emulsified in equal volume of complete Freund's adjuvant (CFA Chondrex Inc. Washington, USA) and administered subcutaneously in a volume of 200 µl, twice over two weeks on opposite flanks. A week after the second immunization, mice were anesthetized by i.p. injection with 9.75 mg xylasol and 48.75 mg ketasol per kg (Dr. E. Graeub AG, Bern, Switzerland) and then given intranasal challenges of 15 µg of HDM in a volume of 30ul PBS on 4 consecutive days. Analysis was performed one day after the final challenge.

[0137] Preparation and administration of positive control compound anti mouse IL-17 antibody . Anti-IL-17 neutralizing antibody was sourced from Bio X Cell and was stored at 4°C (Clone 17F3, Cat. Number BE0173, Bio X Cell) and administered per i.p. at a dose of 12.5 mg / kg at days indicated in study protocol above.

[0138] Terminal procedure. On day 18 animals were sacrificed by lethal i.p. injection with pentabarbitol (Streuli Pharma AG, Uznach, Cat: 117013 9A) immediately followed by bronchoalveolar lavage (BAL) in 500 ul of saline.

[0139] Measurement of cellular infiltrates into BAL. Cells were isolated from the BAL fluid and differential cell counts were performed based upon standard morphological and cytochemical criteria.

[0140] Graphs and statistical analysis. All graphs were generated with Graphpad Prism Version 6 and a one-way ANOVA was applied. Results from the statistical analysis are provided with the individual data tables. Error bars represent Standard Error of the Mean (SEM).Results and analysis

[0141] The results of the experiment are shown in Figures 10-18.

[0142] No morbidity or mortality was noted in the mice treated with the bacteria or the vehicle. As shown in Figures 15 and 16, strain 751 was highly efficacious in alleviating the magnitude of the neutrophilic inflammatory response. Indeed, the treatment with strain 751 showed comparable results to treatment with anti-IL-17 antibodies. In addition, strain 751 reduced eosinophil numbers relative to the controls, as shown in Figures 11 and 12.Example 3 - Efficacy of bacterial inocula to treat arthritis in a type II collagen-induced arthritis mouse model Materials and methodsStrains 751: bacterium deposited under accession number NCIMB 42380Bacterial cultures

[0143] Bacterial cultures were grown up for administration in an anaerobic workstation (Don Whitley Scientific).

[0144] Bacterial strain #751 was grown using glycerol stocks. The glycerol stocks were stored at -80°C. Three times per week, glycerol stocks were thawed at room temperature and streaked on YCFA plates. A new glycerol aliquot was used on each occasion. Bacteria were allowed to grow on a given plate for up to 72 hours.

[0145] Solutions to be administered to the animals were prepared twice daily with an eight hour interval for morning (AM) and afternoon (PM) treatments. A bacterial colony was picked from the streaked plate and transferred into a tube containing YCFA media. Bacterial strain #751 was allowed to grow for 16 hours before AM administrations. Bacteria were sub-cultured at 1% into YCFA media for PM administrations. OD values were recorded for each strain after morning and afternoon treatment preparations.Type II collagen-induced arthritis mouse model

[0146] Adult male DBA / 1 mice were randomly allocated to experimental groups and allowed to acclimatise for two weeks. On Day 0, animals were administered by subcutaneous injection with 100 microliters of an emulsion containing 100 micrograms of type II collagen (CII) in incomplete's Freund's adjuvant supplemented with 4 mg / ml Mycobacterium tuberculosis H37Ra. On Day 21, animals were administered by subcutaneous injection with a booster emulsion containing 100 µg of type II collagen in incomplete Freund's adjuvant.

[0147] Treatments were given according to the administration schedule below. From Day -14 until the end of the experiment on Day 45, animals were weighed three times per week. From Day 21 until the end of the experiment, animals were scored three times per week for clinical signs of arthritis to include swelling of the hind- and front paws, radio-carpal (wrist) joints and tibio-tarsal (ankle) joints.

[0148] On Day 45 mice were culled and terminal blood samples were taken for cytokine analysis.

[0149] On Day -14, Day 0 and Day 45, faecal samples were collected for microbiological analysis, immediately snap-frozen and stored at -80°C.

[0150] The collagen-induced arthritis (CIA) mouse model is a well-established mouse model for rheumatoid arthritis

[71] . Immunisation with CII causes a pathogenesis that includes several important pathological features of rheumatoid arthritis, including synovial hyperplasia, mononuclear cell infiltration and cartilage degradation. Significantly, the development of CIA is mediated by Th17 cells through secretion of IL-17A

[72] . The immune response underlying the arthritis model is enhanced by the use of Freund's adjuvant supplemented with Mycobacterium tuberculosis.

[0151] On Day 21, spleens were collected from three satellite animals in each group. Cells were cultured for 72 hours in the presence or absence of type II collagen. Cytokines, including TNF-a, IL-6, IFN-γ, IL-4, IL-10 and IL-17, were quantified in the culture supernatants and in terminal serum by Luminex. Cell proliferation was quantified using a tritiated thymidine incorporation method.Treatment Groups and Dosages All Groups were n=15 (n=12 for the main study group and n=3 for satellite groups)

[0152] The vehicle used for the biotherapeutics was Yeast extract-Casitone-Fatty Acids (YCFA) medium. Group Dose Administration Disease Induction Route Regimen 1Vehicle5 ml / kgPOBID: Day -14-EndDay 0: Collagen / CFA, once, SC2Biotherapeutic #7515 ml / kgPOBID: Day -14-EndDay 21: Collagen / IFA, once, SCPO: oral gavage, SC: subcutaneous injection, BID: twice a day, CFA: complete Freund's adjuvant. Bodyweights

[0153] From Day -14 until the end of the experiment, animals were weighed three times per week. Data were graphed (Mean ± SEM).Non-specific clinical observations

[0154] From Day -14 until the end of the experiment, animals were checked daily for non-specific clinical signs to include abnormal posture (hunched), abnormal coat condition (piloerection) and abnormal activity levels (reduced or increased activity).Clinical Observations

[0155] From Day 21 until the end of the experiment on Day 45, animals were scored three times per week for clinical signs of arthritis to include swelling of the hind- and front paws, radio-carpal (wrist) joints and tibio-tarsal (ankle) joints. Each limb was scored using the following scale: (0) normal, (1) slight swelling, (2) mild swelling, (3) moderate swelling and (4) severe swelling. A clinical score was calculated by adding each limb score. The maximum possible clinical score for an animal was (16). Animals with a score equal to (12) on two consecutive occasions and animals with a score greater than (12) on any one occasion were culled. Data were graphed (Mean ± SEM).Cell proliferation analysis

[0156] On Day 21, three satellite animals per group were culled and spleens were dissected out. Spleen cells were cultured for 72 hours in presence or absence of type II Collagen. After 72 hours, cells were pulsed overnight in the presence of tritiated thymidine. Cell proliferation was quantified by measuring thymidine incorporation. Data were graphed (Mean ± SEM). Supernatants were taken and tested for the presence of key cytokines.Cytokine analysis

[0157] Terminal supernatants from the spleen cell cultures were tested in order to quantitate TNF-α, IL-6, IFN-γ, IL-4, IL-10 and IL-17 by Luminex. Data were graphed (Mean ± SEM).Microbiological analysis

[0158] On Day -14, Day 0 and Day 45, faecal samples were collected from each animal, immediately snap-frozen, and stored at -80°C. Caeca (including content) were immediately snap-frozen and stored at -80°C. A bacterial identification test was performed daily by plating the bacteria.Histopathology

[0159] At the end of the experiment, hind paws were stored in tissue fixative. Samples were transferred into decalcification solution. Tissue samples were processed, sectioned and stained with Haematoxylin & Eosin. Sections were scored by a qualified histopathologist, blind to the experimental design, for signs of arthritis to include inflammation, articular cartilage damage and damage to the underlying metaphyseal bone. A detailed scoring system was used (see below). Data were graphed (Mean ± SEM). Raw and analysed data were provided as well as representative pictures. Table 1: Histopathology Scoring System Grade Description Inflammation 0Normal joint1Mild synovial hyperplasia with inflammation dominated by neutrophils. Low numbers of neutrophils and macrophages in joint space.2Synovial hyperplasia with moderate to marked inflammation involving both neutrophils and macrophages. Neutrophils and macrophages in joint space; may be some necrotic tissue debris.3Synovial hyperplasia with marked inflammation involving both neutrophils and macrophages. Loss of synoviocyte lining. Inflammation may extend fromsynovium to surrounding tissue including muscle. Numerous neutrophils and macrophages in joint space, together with significant necrotic tissue debris.Articular cartilage damage 0Normal joint1Articular cartilage shows only mild degenerative change. Early pannus formation may be present peripherally.2Articular cartilage shows moderate degenerative change and focal loss. Pannus formation is present focally.3Significant disruption and loss of articular cartilage with extensive pannus formation.Damage to the underlying metaphyseal bone 0Normal joint1No change to underlying metaphyseal bone.2May be focal necrosis or fibrosis of metaphyseal bone.3Disruption or collapse of metaphyseal bone. Extensive inflammation, necrosis or fibrosis extending to medullary space of the metaphysis. Results and analysisSurvival and Non-specific Clinical Observations

[0160] Some animals were culled prior to the scheduled end of the study due to the severity of the clinical signs of arthritis or due to the severity of the non-specific clinical observations.

[0161] Three animals were culled or found dead or culled during the pre-treatment period (Day -14 to Day 0): one animal in Group 1 (vehicle-treated, animal arrived from supplier with broken leg and was culled) and two animals in Group 2 (biotherapeutic #751-treated, possible lung dosing on first pre-treatment day, and post-dose clinical signs on second pre-treatment day).

[0162] Eight animals were culled due to the severity of the clinical signs of arthritis: five animals in Group 1 (vehicle-treated) and three animals in Group 2 (biotherapeutic #751-treated).

[0163] Four animals were culled due to the severity of the non-specific clinical signs including abnormal posture (hunched), abnormal coat condition (piloerection), abnormal activity levels (reduced activity): three animals in Group 1 (vehicle-treated) and one animal in Group 2 (biotherapeutic #751-treated).Bodyweights

[0164] Bodyweight data recorded from Day -14 until Day 0 and expressed as a percentage of the initial (Day -14) bodyweights were analysed by two-way ANOVA followed by Dunnett's post-test for multiple comparisons with Day -14 then for multiple comparison with the vehicle-treated group. The data are presented in Figure 19. Data from animals culled prior to the scheduled end of the experiment were excluded from the analyses.

[0165] When compared to Day -14, twice daily administrations by oral gavage induced a significant bodyweight loss in the vehicle-treated group on Day -9 and Day -7.

[0166] The bodyweights measured between Day -14 and Day -1 in the biotherapeutic-treated groups did not differ from the bodyweights measured in the vehicle-treated group on any given day.

[0167] Bodyweight data recorded from Day 0 until Day 28 and expressed as a percentage of the initial (Day 0) bodyweights were analysed by two-way ANOVA followed by Dunnett's post-test for multiple comparisons with Day 0 in the Vehicle group then for multiple comparison with the vehicle-treated group. The data are presented in Figure 20. Data from animals culled prior to the scheduled end of the experiment and from Satellite animals were excluded from the analyses. Day 28, Day 35 and Day 42 data were further analysed by one-way ANOVA followed by Dunnett's post-test for multiple comparisons to the vehicle-treated group.

[0168] The onset of clinical signs of arthritis was associated with a significant bodyweight loss on Day 26 and Day 28 (p < 0.0001) when compared to Day 0 in the vehicle-treated group.

[0169] There was no significant difference between experimental groups on Day 35 or Day 42.Clinical Observations

[0170] Clinical score data were analysed by two-way ANOVA followed by Dunnett's post-test for multiple comparisons between days in the vehicle-treated group then for multiple comparisons between experimental groups and the vehicle-treated group each day. The data are presented in Figure 21. Data recorded from animals culled prior to the end of the experiment were excluded from the analysis. When animals were culled due to the severity of the clinical signs of arthritis, the last recorded score was reported for the following days and used in the statistical analyses.

[0171] A significant increase of the clinical scores was observed in the vehicle-treated group from Day 28 until Day 45 (p < 0.0001) when compared to Day 21.

[0172] Biotherapeutic #751 induced a reduction of the clinical scores when compared to the vehicle-treated group from Day 31 until Day 45, although the difference was non-significant.Cell proliferation analysis

[0173] To validate the assay, splenocytes were cultured in the presence of soluble anti-CD3 and anti-CD28 (anti-CD3 / CD28) as positive control stimuli to confirm the proliferative potential of the cells.

[0174] Strong proliferative responses to anti-CD3 / CD28 were seen in all experimental groups, showing cells were healthy, viable and able to respond to activation signals.

[0175] To test the proliferative response in presence of Collagen II (CII), splenocytes were cultured in the presence of CII at 50 µg / ml. Splenocyte proliferative response to CII were analysed by two-way ANOVA followed by Sydak's post-test for multiple comparisons between unstimulated and CII-stimulated splenocytes and one-way ANOVA followed by Dunnett's post-test for comparison of CII-stimulated response in different experimental groups with the vehicle-treated group. The data are presented in Figure 22.

[0176] CII induced a highly significant increase of 3< H-thymidine incorporation (cpm) when compared to the unstimulated splenocytes in the vehicle-treated group (p < 0.0001).

[0177] The groups treated with biotherapeutic #751demonstrated significantly lower levels of CII-induced splenocyte proliferation than the vehicle-treated group.Cytokine levels in tissue culture supernatants

[0178] Levels of each cytokine were measured in tissue culture supernatants derived from anti-CD3 / CD28 stimulated cultures by luminex analysis. These showed robust responses for all cytokines measured (mean levels in vehicle group were as follows: IL-4 = 6,406 pg / ml; IL-6 = 306 pg / ml; IL-10 = 10,987 pg / ml; IL-17A = 11,447 pg / ml; IFN-γ = 15,581 pg / ml; TNF-α = 76 pg / ml).

[0179] The following sections summarise the data obtained from the Collagen II-stimulated cultures. Where applicable, statistical analyses of the differences between cytokine levels in supernatants of unstimulated and CII-stimulated splenocytes were conducted using two-way ANOVA followed by Sidak's post-test for multiple comparisons, while one-way ANOVA followed by Dunnett's post-test was used for comparison of CII-stimulated response in biotherapeutic-treated groups with the vehicle-treated group. There was no significant difference in cytokine levels between the groups in both cases. This is likely due to the small sample size used (n=3).

[0180] In order to more accurately present the distribution of the data for the cytokines with substantial spread of the data, these are presented as scatter plots.

[0181] The group means of IL-4 in tissue culture supernatants after stimulation with CII were <5pg / ml. These are not considered biologically significant and not included here. The group means of TNF-α in tissue culture supernatants after stimulation with collagen were below limit of quantitation.Supernatant levels of IFN-γ (Figure 23)

[0182] Along with IL-17, IFN-γ is the major cytokine driving disease in the CIA model. The scatter plot in Figure 23 demonstrates IFN-γ levels after CII stimulation, with group median being higher for the Vehicle-treated group compared to the biotherapeutic. The outlier result from the same group 2 subject is responsible for the higher median in this group for IFN-γ and IL-10.Supernatant levels of IL-17A (Figure 24)

[0183] Levels of IL-17A were 50pg / ml in CII-stimulated cultures for the Vehicle-treated group. The levels of this cytokine appeared to be lower in the biotherapeutic group compared to the Vehicle-treated.Supernatant levels of IL-10 (Figure 25)

[0184] Levels of IL-10 in Vehicle-treated group were 13 pg / ml and 2.1 pg / ml for CII-stimulated, and media control cultures, respectively. Higher levels of IL-10 (which is an anti-inflammatory cytokine) for the vehicle-treated group may be expected because inflammation and pro-inflammatory cytokine induction could be accompanied by an anti-inflammatory feedback mechanism.Supernatant levels of IL-6 (Figure 26)

[0185] Inflammatory cytokines such as IL-6 and TNF-α are not typically produced at high levels in anti-CII cultures. However, their levels may be altered as a result of immune modulation. Levels of IL-6 in CII-stimulated cultures were modest, reaching 10pg / ml. Although higher than in media control cultures, these differences were too small to provide rationale for performing statistical analyses.Microbiological analysis

[0186] Bacterial growth was confirmed by measuring the optical density at 600 nm using a spectrophotometer. Bacterial identity was confirmed by comparing streaked plate pictures to reference pictures.

[0187] Following the improved bacterial preparation method, consistently high doses of bacterial strain were administered from Day -2 and Day -3 as indicated by the high OD values measured.

[0188] Faecal samples were collected and snap-frozen on Day -14, Day 0 and at termination.Histopathology

[0189] The histopathology results are shown in Figures 66-70. As expected for this model, intra-individual and inter-individual variability was observed in terms of the presence / absence of arthritis or the severity of change present.

[0190] The nature of the pathology was as expected for this model, with extensive mixed chronic-active inflammation of the synovium and bursa extending to involve the peri-articular soft tissues (muscle, adipose tissue, dermal collagen). In the most severely affected joints there was articular cartilage degeneration and loss with intra-articular debris and inflammation and disruption of the joint and bone structure by fibrosis and inflammation.

[0191] The incidence of histopathological changes was: vehicle - 80% (16 / 20); Biotherapeutic #751 - 45% (9 / 20). Treatment with Biotherapeutic #751 reduced the incidence of histopathological scores in mouse hind limbs when compared to the vehicle-treated group (see Figures 66-69). Histopathology scores were analysed by one-way ANOVA for non-parametric data (Kruskal-Wallis test) followed by Dunn's post-test for multiple comparisons to the vehicle-treated group. Biotherapeutic #751 induced a significant reduction of the joint inflammation scores observed in histopathology when compared to the vehicle-treated group (p < 0.01). Biotherapeutic #751 induced a significant reduction of the cartilage damage scores observed in histopathology when compared to the vehicle-treated group (p < 0.001). Biotherapeutic #751 induced a significant reduction of the bone damage scores observed in histopathology when compared to the vehicle-treated group (p < 0.001). Biotherapeutic #751 induced a significant reduction of the total histopathology scores when compared to the vehicle-treated group (p < 0.01).Summary

[0192] Increased clinical scores were observed from Day 28 after the first administration of type II collagen, as expected in this model of arthritis in DBA / 1 mice. Biotherapeutic #751 was shown to be effective at treating arthritis in this model and Biotherapeutic #751 was effective for reducing the severity of the clinical scores. Biotherapeutic #751 was also effective for reducing pathological disease in the joints, as demonstrated in the histopathological analysis.

[0193] Proliferative recall responses to Collagen II were seen in splenocyte cultures from all experimental groups. The collagen-specific response was significantly reduced following treatment with biotherapeutic #751 (Group 2).

[0194] Most of the T cell cytokines tested showed detectable increases between Collagen II-stimulated and media controls in the Vehicle-treated group. These increases were not as obvious in the biotherapeutic-treated group. This broadly supports the proliferative recall responses to Collagen II described above.

[0195] There was evidence of suppression of the Th1 / Th17 axis, which is the pathogenic response in this model and in human RA. Correlation of reduced levels of cytokines with reduced proliferation is suggestive of immune modulation. There was no evidence that this modulation resulted either from enhanced levels of Th2 associated IL-4 or with increases in the immune modulating cytokine, IL-10.Example 4 - Further analysis of the effect of bacterial inocula in the mouse model of house dust mite-induced asthma

[0196] The mice tested in Example 1 were subjected to further analyses to further characterise the effect of the compositions of the invention on the allergic asthma inflammatory response.Materials and methods

[0197] Blood withdrawal and serum preparation on day 14. Blood samples of animals were collected via cardiac puncture. Serum was isolated from the blood sample by centrifugation for 5 min at 14000g and stored at -20 °C.

[0198] Organ removal on day 14. Collection of the left lung lobe in formalin for follow-on histological analysis. Collection of the right lung lobes (all remaining lobes) and removal of serum for snap freezing and follow-on analysis. Remaining BAL fluid was snap frozen for follow-on analysis.Measurement of antibody levels in serum and BAL fluid

[0199] Total IgE and house-dust-mite (HDM) specific IgG1 antibody production were measured in the BAL and serum by ELISA assay.Isolation of lung and histological analysis

[0200] Left lung lobes were fixed in formalin followed by embedment in paraffin, sectioning, and staining with hematoxylin and eosin and PAS. Subsequent histological scoring was performed blinded as followed: Five random fields of view per sample were scored for inflammation (peribronchial infiltration and perivascular infiltration) and mucus production. Inflammatory infiltration was scored with the following grading system: 0 - normal 1 - mild inflammatory infiltrates 2 - moderate inflammatory infiltrates 3 - marked inflammatory infiltrates 4 - severe inflammatory infiltrates 5 - very severe inflammatory infiltrates

[0201] In each field of view, airways were measured in size and mucus cell numbers were quantified / um.Measurement of inflammatory mediators in lung tissue

[0202] Right lung lobes (all remaining lobes) isolated for quantification of inflammatory mediators were snap frozen for subsequent measurement of CCL11, IFN-gamma, IL-1 alpha, IL-1 beta, IL-4, IL-5, IL-9, IL-17A, CXCL1, CCL3, CXCL2 and CCL5 by commercially available multiplex assay (Merck-Millipore). Analysis was performed according to the manufacturer's instructions.Results and analysis

[0203] The results of the experiments are shown in Figures 28-46.

[0204] In support of the findings described in Example 1, analysis of the cellular infiltrates in the lung tissue of mice treated with strain 751 showed a notable and statistically significant reduction in mean inflammation score (see Figures 32 and 34).

[0205] Antibody levels in the BAL fluid and serum were analysed (see Figures 28-31). No clear effect of the bacterial treatment on serum antibody levels was observed. This may reflect a failure in the experiment, because the spread of data and the error bars for each treatment are large, and the positive and negative controls do not appear to have behaved as would be expected. Also, the baseline serum antibody levels could have masked any changes.

[0206] Similarly, no clear effect of the bacterial treatment on cytokine levels in lung tissue was observed (see Figures 36-46). Again, this may reflect a failure in the experiment, because the spread of data and the error bars for each treatment are large, and the positive and negative controls do not appear to have behaved as would be expected. It is also possible that the mechanism of action involved influences earlier cytokine responses that were no longer detectable on day 4 post the final HDM airway challenge. Some care should be taken when interpreting the cytokine data in the current study, due to the variability in the levels detected. This variability could in part be explained by the fact that the lung tissue was separated for the different analyses, and thus one lung lobe might not have been fully representative or comparable to the same lobe in other mice due to patchy distribution of the inflammation.Example 5 - Further analysis of the effect of bacterial inocula in the mouse model of severe neutrophilic asthma

[0207] The mice tested in Example 2 were subjected to further analyses to further characterise the effect of the compositions of the invention on the neutrophilic response associated with severe asthma.Materials and methods

[0208] Organ removal on day 18. Collection of the left lung lobe in formalin for follow-on histological analysis. Collection of the right lung lobes (all remaining lobes) and removal of serum for snap freezing and follow-on analysis. Remaining BAL fluid was snap frozen for follow-on analysis.

[0209] Measurement of inflammatory mediators in lung tissue (follow-on analysis). Right lung lobes (all remaining lobes) isolated for quantification of inflammatory mediators were snap frozen for subsequent measurement of IFN-gamma, IL-1 alpha, IL-1 beta, CXCL1, CCL3, CXCL2, CCL5, IL-17A, TNF-alpha, IL-17F, IL-23 and IL-33 by commercially available multiplex assay (Merck-Millipore). Analysis was performed according to the manufacturer's instructions.

[0210] Measurement of antibody levels in serum and BAL fluid (follow-on analysis). House-dust-mite (HDM) specific IgG1 and IgG2a antibody production were measured in the BAL and serum by ELISA assay.

[0211] Isolation of lung and histological analysis (follow-on analysis). Left lung lobes were fixed in formalin followed by embedment in paraffin, sectioning, and staining with hematoxylin and eosin and PAS. Subsequent histological scoring was performed blinded as followed: Five random fields of view per sample were scored for inflammation (peribronchial infiltration and perivascular infiltration) and mucus production. Inflammatory infiltration was scored with the following grading system: 0 - normal 1 - mild inflammatory infiltrates 2 - moderate inflammatory infiltrates 3 - marked inflammatory infiltrates 4 - severe inflammatory infiltrates 5 - very severe inflammatory infiltrates Results and analysis

[0212] The results of the experiments are shown in Figures 47-64.

[0213] Further analysis of antibody levels revealed that the efficacy of bacterial strain 751 was also reflected in reduced HDM-specific IgG1 levels in the BAL fluid and serum (see Figures 47 and 49). Firm conclusions regarding an effect on IgG2a levels cannot be drawn. Overall, the data from the antibody analysis is suggestive of a reduction related to an overall reduced inflammatory response, as opposed to a selective effect on antibody isotype switching.

[0214] Histological analysis supported the differential cell counts from the BAL fluid, showing a reduced cellular infiltrate in mice treated with Strain 751 (see Figures 51-53).

[0215] In relation to cytokine levels, as for Example 4, the spread of data and the error bars for each treatment are large, and the positive and negative controls do not appear to have behaved as necessarily would be expected. It is also possible that the mechanism of action involves influencing earlier cytokine responses that were no longer detectable on day 4 post the final HDM airway challenge. Some care should be taken when interpreting the cytokine data in the current study, due to the variability in the levels detected. This variability could in part be explained by the fact that the lung tissue was separated for the different analyses, and thus one lung lobe might not have been fully representative or comparable to the same lobe in other mice due to patchy distribution of the inflammation. Despite this variability, a clear anti-inflammatory effect on cytokine levels for strain 751 was shown, and the positive control anti-IL-17 Ab generally behaved as expected.

[0216] With the above caveats, the data in Figures 56, 58, 59, 61 and 63 suggest that treatment with the bacterial strains of the invention, and in particular strain 751 may achieve a reduction in the levels of IL-1b, IFNg, RANTES, MIP-1a and KC (the mouse orthologue of human IL-8), which may be indicative of a mechanism of action related to influences on chemokine release (and thus recruitment of cells) by stromal or innate immune cells. These cytokines are part of the Th17 pathway. Taking this dataset together, a clear conclusion can be drawn that Strain 751 was highly effective at protecting mice against inflammation in this mouse model of severe neutrophilic asthma.Example 6 - Efficacy of bacterial inocula in a mouse model of multiple sclerosis Summary

[0217] Mice were administered with compositions comprising bacterial strains according to the invention and the mice were subsequently immunised with myelin oligodendrocyte glycoprotein to induce experimental autoimmune encephalomyelitis (EAE). EAE is the most commonly used experimental model for human multiple sclerosis. The compositions of the invention were found to have a striking effect on disease incidence and disease severity.Strain751: bacterium deposited under accession number NCIMB 42380Study designGroups:

[0218] 1. Negative control group. Treatment with vehicle control (per oral). 4. Treatment with therapeutic bacteria inoculum strain 751 (per oral). 9. Positive control group. Treatment with Dexamethasone (i.p.). 10. Untreated Control Group. Number of mice per group = 10 Days -14 to day 27: Daily administration of vehicle control per oral (Group 1). Days -14 to day 27: Daily administration of therapeutic bacteria inoculum per oral (Group 4). Days 0-28: administration of Dexamethasone (i.p.) three times a week (Group 9) Day 0: MOG35-55 (myelin oligodendrocyte glycoprotein - 2mg / ml) and CFA (2mg / ml MTB) were mixed 1:1 resulting in 1mg / ml solutions. 100µl of the peptide-CFA mixture was injected subcutaneously into each hind leg. Administration of pertussis toxin intraperitoneally (300ng). Day 1: Administration of pertussis toxin intraperitoneally (300ng). Days 7-onwards: Measurement of disease incidence and weight three times a week. Endpoints and analysis

[0219] Mice were analysed for disease incidence and disease severity three times a week. Scoring was performed blind. Disease severity was assessed using a clinical score ranging from 0 to 5, with 5 indicating a dead mouse (see clinical scoring system below).Monitoring

[0220] On the indicated days mice were weighed and observed for disease activity score and disease incidence.

[0221] Disease activity score observations: 0 -No obvious changes in motor function compared to non-immunized mice. 0.5 -Tip of tail is limp. 1.0 -Limp tail. 1.5 -Limp tail and hind leg inhibition. 2.0 -Limp tail and weakness of hind legs. OR - There are obvious signs of head tilting when the walk is observed. The balance is poor. 2.5 -Limp tail and dragging of hind legs. - OR - There is a strong head tilt that causes the mouse to occasionally fall over. 3.0 -Limp tail and complete paralysis of hind legs. 3.5 -Limp tail and complete paralysis of hind legs.

[0222] In addition to: Mouse is moving around the cage, but when placed on its side, is unable to right itself.

[0223] Hind legs are together on one side of body. 4.0 -Limp tail, complete hind leg and partial front leg paralysis. - Mouse is minimally moving around the cage but appears alert and feeding 4.5 -Complete hind and partial front leg paralysis, no movement around the cage. Mouse is immediately euthanized and removed from cage. 5.0Mouse is euthanized due to severe paralysis.

[0224] When an animal has equal or greater disease activity score of 1, it is considered to have a positive disease incidence score.Results

[0225] The results of the study are shown in Figures 71 and 72.

[0226] Disease induction in the negative control groups was successful with high scores shown by the vehicle control and the untreated control. The effect of treatment with strain 751 was striking and the mice treated with strain 751 exhibited notably reduced disease incidence and disease severity. Indeed, the reduction in disease incidence and disease severity was comparable to the positive control group. These data indicate the strain 751 may be useful for treating or preventing multiple sclerosis.Example 7 - Efficacy of bacterial inocula in mouse models of cancer Summary

[0227] This study tested the efficacy of compositions comprising bacterial strains according to the invention in four tumor models.Materials

[0228] Test substance - Bacterial strain #MRX004 (strain 751).

[0229] Reference substance - Anti-CTLA-4 antibody (clone: 9H10, catalog: BE0131, isotype: Syrian Hamster IgG1, Bioxcell).

[0230] Test and reference substances vehicles - Bacterial culture medium (Yeast extract, Casitone, Fatty Acid medium (YCFA)). Each day of injection to mice, antibody was diluted with PBS (ref: BE14-516F, Lonza, France).

[0231] Treatment doses - Bacteria: 2×10 8< in 200 µL. The a-CTLA-4 was injected at 10 mg / kg / inj. Anti-CTLA-4 was administered at a dose volume of 10 mL / kg / adm (i.e. for one mouse weighing 20 g, 200 µL of test substance will be administered) according to the most recent body weight of mice.

[0232] Routes of administration - Bacterial inoculum was administered by oral gavage (per os, PO) via a cannula. Cannulas were decontaminated every day. Anti-CTLA-4 was injected into the peritoneal cavity of mice (Intraperitoneally, IP).

[0233] Culture conditions of bacterial strain - The culture conditions for the bacterial strain were as follows: Pipette 10 mL of YCFA (from the prepared 10 mL E&O lab bottles) into Hungate tubes Seal the tubes and flush with CO 2 using a syringe input and exhaust system Autoclave the Hungate tubes When cooled, inoculate the Hungate tubes with 1 mL of the glycerol stocks Place the tubes in a static 37°C incubator for about 16 hours. The following day, take 1 mL of this subculture and inoculate 10 mL of YCFA (pre-warmed flushed Hungate tubes again, all in duplicate) Place them in a static 37°C incubator for 5 to 6h Cancer cell line and culture conditions -

[0234] The cell lines that were used are detailed in the table below: Cell line Type Mouse strain Origin EMT-6Breast carcinomaBALB / cATCCLL / 2 (LLC1)Lung carcinomaC57BL / 6ATCC CRL1642Hepa1-6Hepatocellular carcinomaC57BL / 6IPSEN INNOVATION

[0235] The EMT-6 cell line was established from a transplantable murine mammary carcinoma that arose in a BALB / cCRGL mouse after implantation of a hyperplastic mammary alveolar nodule

[73] .

[0236] The LL / 2 (LLC1) cell line was established from the lung of a C57BL mouse bearing a tumor resulting from an implantation of primary Lewis lung carcinoma

[74] .

[0237] The Hepa 1-6 cell line is a derivative of the BW7756 mouse hepatoma that arose in a C57 / L mouse

[75] .

[0238] Cell culture conditions - All cell lines were grown as monolayer at 37°C in a humidified atmosphere (5% CO 2 , 95% air). The culture medium and supplement are indicated in the table below: Cell line Culture medium Supplement EMT6RPMI 1640 containing 2mM L-glutamine (ref: BE12-702F, Lonza)10% fetal bovine serum (ref: #3302, Lonza)LL / 2 (LLC1)RPMI 1640 containing 2mM L-glutamine (ref: BE12-702F, Lonza)10% fetal bovine serum (ref: #3302, Lonza)Hepa1-6DMEM (ref: 11960-044, Gibco)10% fetal bovine serum (ref: #3302, Lonza)2mM L-Glutaminepenicillin-streptomycin (Sigma G-6784)

[0239] For experimental use, adherent tumor cells were detached from the culture flask by a 5 minute treatment with trypsin-versene (ref: BE17-161E, Lonza), in Hanks' medium without calcium or magnesium (ref: BE10-543F, Lonza) and neutralized by addition of complete culture medium. The cells were counted in a hemocytometer and their viability will be assessed by 0.25% trypan blue exclusion assay.Use of animals -

[0240] Healthy female Balb / C (BALB / cByJ) mice, of matching weight and age, were obtained from CHARLES RIVER (L'Arbresles) for the EMT6 model experiments.

[0241] Healthy female C57BL / 6 (C57BL16J) mice, of matching weight and age, were obtained from CHARLES RIVER (L'Arbresles) for the LL / 2(LLC1) and the Hepa1-6 model experiments.

[0242] Animals were maintained in SPF health status according to the FELASA guidelines, and animal housing and experimental procedures according to the French and European Regulations and NRC Guide for the Care and Use of Laboratory Animals were followed [76,77]. Animals were maintained in housing rooms under controlled environmental conditions: Temperature: 22 ± 2°C, Humidity 55 ± 10%, Photoperiod (12h light / 12h dark), HEPA filtered air, 15 air exchanges per hour with no recirculation. Animal enclosures were provided with sterile and adequate space with bedding material, food and water, environmental and social enrichment (group housing) as described: 900 cm 2< cages (ref: green, Tecniplast) in ventilated racks, Epicea bedding (SAFE), 10 kGy Irradiated diet (A04-10, SAFE), Complete food for immuno-competent rodents - R / M-H Extrudate, water from water bottles.Experimental design and treatmentsAntitumor activity, EMT6 model

[0243] Treatment schedule - The start of first dosing was considered as D0. On D0, non-engrafted mice were randomized according to their individual body weight into groups of 9 / 8 using Vivo manager ®< software (Biosystemes, Couternon, France). On D0, the mice received vehicle (culture medium) or bacterial strain. On D14, all mice were engrafted with EMT-6 tumor cells as described below. On D24, mice from the positive control group received anti-CTLA-4 antibody treatments.

[0244] The treatment schedule is summarized in the table below: Group No. Animals Treatment Dose Route Treatment Schedule 18Untreated---28Vehicle (media)-POQ1Dx4239Bacterial strain #1 (MRX004)2×108 bacteriaPOQ1Dx4248Anti-CTLA410 mg / kgIPTWx2

[0245] The monitoring of animals was performed as described below.

[0246] Induction of EMT6 tumors in animals - On D14, tumors were induced by subcutaneous injection of 1×10 6< EMT-6 cells in 200 µL RPMI 1640 into the right flank of mice.

[0247] Euthanasia - Each mouse was euthanized when it reached a humane endpoint as described below, or after a maximum of 6 weeks post start of dosing.Antitumor activity, LL / 2 (LLC1) model

[0248] Treatment schedule - The start of first dosing was considered as D0. On D0, non-engrafted mice were randomized according to their individual body weight into 7 groups of 9 / 8 using Vivo manager ®< software (Biosystemes, Couternon, France). On D0, the mice will received vehicle (culture medium) or bacterial strain. On D14, all mice were engrafted with LL / 2 tumor cells as described below. On D27, mice from the positive control group received anti-CTLA-4 antibody treatments.

[0249] The treatment schedule is summarized in the table below: Group No. Animals Treatment Dose Route Treatment Schedule 18Untreated---29Vehicle (media)-POQ1Dx4239Bacterial strain # 1 (MRX004)2×10 8< bacteriaPOQ1Dx4248Anti-CTLA410 mg / kgIPTWx2

[0250] The monitoring of animals was performed as described below.

[0251] Induction of LL / 2 (LLC1) tumors in animals - On D14, tumors were induced by subcutaneous injection of 1×10 6< LL / 2 (LLC1) cells in 200 µL RPMI 1640 into the right flank of mice.

[0252] Euthanasia - Each mouse was euthanized when it reached a humane endpoint as described below, or after a maximum of 6 weeks post start of dosing.Antitumor activity, Hepa1-6 model

[0253] Treatment schedule - The start of first dosing was considered as D0. On D0, non-engrafted mice were randomized according to their individual body weight into 7 groups of 9 using Vivo manager ®< software (Biosystemes, Couternon, France). On D0, the mice received vehicle (culture medium) or bacterial strain. On D14, all mice were engrafted with Hepa 1-6 tumor cells as described below. On D16, mice from the positive control group received anti-CTLA-4 antibody treatments.

[0254] The treatment schedule is summarized in the table below: Group No. Animals Treatment Dose Route Treatment Schedule 19Untreated---29Vehicle (media)-POQ1Dx4249Bacterial strain #2 (MRX004)2×10 8< bacteriaPOQ1Dx4279Anti-CTLA410 mg / kgIPTWx2

[0255] The monitoring of animals was performed as described below.

[0256] Orthotopic induction of Hepa 1-6 tumor cells in animals by intrasplenic injection - On D14, one million (1×10 6< ) Hepa 1-6 tumor cells in 50 µL RPMI 1640 medium were transplanted via intra-splenic injection into mice. Briefly, a small left subcostal flank incision was made and the spleen was exteriorized. The spleen was exposed on a sterile gauze pad, and injected under visual control with the cell suspension with a 27-gauge needle. After the cell inoculation, the spleen was excised.

[0257] Euthanasia - Each mouse was euthanized when it reached a humane endpoint as described in section below, or after a maximum of 6 weeks post start of dosing.

[0258] Evaluation of tumor burden at euthanasia - At the time of termination, livers were collected and weighed.Animal monitoring

[0259] Clinical monitoring - The length and width of the tumor was measured twice a week with callipers and the volume of the tumor was estimated by this formula

[78] : Tumor volume = width 2 × length 2

[0260] Humane endpoints

[79] : Signs of pain, suffering or distress: pain posture, pain face mask, behaviour; Tumor exceeding 10% of normal body weight, but non-exceeding 2000 mm 3< ; Tumors interfering with ambulation or nutrition; Ulcerated tumor or tissue erosion; 20% body weight loss remaining for 3 consecutive days; Poor body condition, emaciation, cachexia, dehydration; Prolonged absence of voluntary responses to external stimuli; Rapid laboured breathing, anaemia, significant bleeding; Neurologic signs: circling, convulsion, paralysis; Sustained decrease in body temperature; Abdominal distension.

[0261] Anaesthesia - Isoflurane gas anesthesia were used for all procedures: surgery or tumor inoculation, i.v. injections, blood collection. Ketamine and Xylazine anesthesia were used for stereotaxia surgical procedure.

[0262] Analgesia - Carprofen or multimodal carprofen / buprenorphine analgesia protocol were adapted to the severity of surgical procedure. Non-pharmacological care was provided for all painful procedures. Additionally, pharmacological care not interfering with studies (topic treatment) were provided at the recommendation of the attending veterinarian.

[0263] Euthanasia - Euthanasia of animals was performed by gas anesthesia over-dosage (Isoflurane) followed by cervical dislocation or exsanguination.ResultsAntitumor activity, EMT6 model

[0264] The results are shown in Figure 73. Treatment with the bacterial strain of the invention led to a clear reduction in tumour volume relative to both the negative controls. The positive control also led to a reduction in tumour volume, as would be expected.Antitumor activity, LL / 2 (LLC1) model

[0265] The results are shown in Figure 74. The negative and positive controls do not appear as would be expected, because tumour volume was greater in the mice treated with the positive control than in the negative control groups. Nevertheless, tumour volume in the mice treated with the bacterial strain of the invention was comparable to the positive control group, which is consistent with a useful therapeutic effect.Antitumor activity, Hepa1-6 model

[0266] The results are shown in Figure 75. The untreated negative control does not appear as would be expected, because liver weight was lower in this group than the other groups. However, the vehicle negative control and the positive control groups both appear as would be expected, because mice treated with vehicle alone had larger livers than mice treated with anti-CTLA4 antibodies, reflecting a greater tumour burden in the vehicle negative control group. Treatment with the bacterial strain of the invention led to a clear reduction in liver weight (and therefore tumour burden) relative to the mice in the vehicle negative control group.

[0267] These data indicate that strain 751 / MRX004 may be useful for treating or preventing cancer, and in particular for reducing tumour volume in breast, lung and liver cancers.Example 8 - Attachment to human cells in YCFA medium Summary

[0268] The level of binding of strain 751 and a number of Bifidobacterium breve strains to human cells was determined at 3 distinct time points in YCFA medium. The bacteria attached to the human cells were resuspended in medium and the optical density of the medium was then analysed - the higher the optical density, the higher the number of bacterial cells and thus, the higher the level of binding of the bacterial cells to human cells. The 751 strain was found to display reduced attachment to human cells compared to the Bifidobacterium breve reference strains.Results and analysis

[0269] The results of the experiment are shown in Figure 76.

[0270] As shown in Figure 76, the Bifidobacterium breve strains show a high level of attachment to human cells at all time points. On the other hand, the 751 strain has a drastically reduced level of attachment to human cells. Therefore, the low adherence to human cells of strain 751 may increase the beneficial effect of the compositions of the invention on the IL-17 or the Th17 pathway and on diseases mediated by IL-17 or the Th17 pathway.Example 9 - Assay detecting the production of exopolysaccharides Summary

[0271] The level of exopolysaccharide (EPS) production by the bacterial strain of the invention (751) and a number of Bifidobacterium breve strains was analysed at 37°C for 48 hours and at 30°C for 72 hours. EPSs are polysaccharides produced by certain bacteria which bind to the outside surface of the bacterial cell. The level of EPSs on the surface of bacteria can be determined using a Congo Red assay which binds to the polysaccharides. A higher intensity of Congo Red absorbance indicates a higher concentration of EPSs on the surface of the bacteria. The bacterial strain of the invention was found to produce and bind more EPSs than the Bifidobacterium breve strains.Results and analysis

[0272] The results of this experiment are shown in Figure 77.

[0273] As shown in Figure 77, the bacterial strain of the invention showed a greater Congo Red absorbance than the Bifidobacterium breve strains at both temperatures and time-points. Therefore, the strain of the invention displays greater EPS production and a greater level of extracellular bound EPSs. As the EPSs enable bacteria to bind to mucus and epithelial cells, the bacterial strain of the invention may be useful for competing with pathogenic cells for binding sites on epithelial cells and within mucus membranes. Thus, the bacterial strain of the invention may be useful for modulating the microbiome and treating a number of diseases associated with the microbiome.Example 10 - Stability testing

[0274] A composition described herein containing at least one bacterial strain described herein is stored in a sealed container at 25°C or 4°C and the container is placed in an atmosphere having 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90% or 95% relative humidity. After 1 month, 2 months, 3 months, 6 months, 1 year, 1.5 years, 2 years, 2.5 years or 3 years, at least 50%, 60%, 70%, 80% or 90% of the bacterial strain shall remain as measured in colony forming units determined by standard protocols.Example 11 - MRX004 bound and released exopolysaccharide production assay

[0275] For EPS extraction, MRX004 was cultured in 10 ml YCFA until it reached late exponential phase, when bacterial cells and supernatants were separated by centrifugation. Cells were washed once with PBS to remove any remaining culture medium. Secreted or released EPS (EPS-R) was precipitated from culture supernatants by treatment with ice-cold 100 % ethanol (overnight at 4°C with gentle agitation). To extract capsular or bound EPS (EPS-B), cells were incubated with 0.05 M EDTA (overnight at 4 °C with gentle agitation), and supernatants from this treatment were collected and subsequently treated with 100 % ice-cold ethanol (overnight at 4 °C with gentle agitation) to precipitate EPS-B. Precipitated EPS-B and EPS-R were pelleted by centrifugation and were allowed to dry briefly in a laminar hood before they were resuspended in enough sterile Ultrapure water to achieve a uniform solution. To further purify samples, they were dialysed against sterile Ultrapure water at a ratio of 1:100 for 48 hours with 3 buffer changes. EPS-B and EPS-R were quantified using the phenol-sulphuric acid method using glucose as standard. Using this assay, MRX004 was found to produce a greater amount of EPS-R (115 µg) than EPS-B (17 µg) (Figure 78).Example 12 - Attachment assay of MRX004 to Caco-2 cells

[0276] Attachment of MRX004 to host cells was analysed using an in vitro co-culture assay with Caco-2 intestinal epithelial cells. Caco-2 cells were seeded at a density of 1 × 10 Bacteria were cultured in 10 ml YCFA until they reached late exponential phase, when they were pelleted, washed twice with PBS, and resuspended in antibiotic-free cell culture medium. Bacterial density was adjusted to obtain an approximate multiplicity of infection (MOI) of 10:1 (which was confirmed by plating on YCFA agar using the WASP standard protocol), and MRX004 was co-incubated with Caco-2 cells in anaerobic conditions at 37°C for 2 hours. Medium was subsequently removed, and unbound bacteria were removed by washing Caco-2 cells three times with PBS. Bacteria-bound Caco-2 cells were lysed and removed from the vessel using treatment with 0.1 % Triton X-100, and 50 µl volumes of diluted lysate were plated in YCFA agar using the WASP. Attachment was calculated by counting the numbers of bacteria recovered from lysate and expressing this as a percentage of total bacteria. MRX004 was found to display low-level adherence (0.3 % of total culture) to Caco-2 cells (Figure 79).Example 13 - Characterisation of enzymatic activity

[0277] The Analytical Profile Index (API ®< ) test system consists of strips that contain miniaturised biochemical tests that assay for enzymatic activity in bacterial species. MRX004 (strain 751, the bacterium deposited under accession number NCIMB 42380) was characterised using two API test systems: Rapid ID 32A - This system is designed specifically for anaerobic species and encompasses tests for carbohydrate, amino acid and nitrate metabolism as well as alkaline phosphatase activity; and API ®< 50 CH - This system tests for the fermentation of 49 carbohydrate sources, and can be utilised in conjunction with API ®< CHL Medium for analysis of anaerobic species.

[0278] Rapid ID 32A testing was carried out on bacterial colonies as per manufacturer's instructions. Briefly, bacteria were cultured on YCFA agar for 24 hours at 37 °C in an anaerobic workstation. Colonies were removed from plates using a sterile 5 µl inoculating loop and resuspended in a 2 ml ampoule of API ®< Suspension Medium until a density roughly equivalent to that of McFarland standard No. 4 was achieved. Fifty-five microlitres of bacterial suspension was added to each cupule on a Rapid ID 32A strip, and the urease test was overlayed with two drops of mineral oil. Strips were covered with a plastic lid and incubated aerobically at 37 °C for 4 hours, following which the bottom row of cupules were developed using the following reagents: NIT: 1 drop each of NIT1 and NIT2; IND: 1 drop of James reagent; all remaining cupules: 1 drop of FastBlue reagent. Strips were incubated at room temperature for 5 minutes, following which the colour of each cupule was recorded and assigned a value of negative, intermediate positive or positive.

[0279] The results of the Rapid ID 32A analysis are shown in Figure 80. MRX004 tested positive for fermentation of several carbohydrate sources, namely α-galactosidase and β-galactosidase, α-glucosidase and β-glucosidase, α-arabinose, mannose and raffinose, as well as the amino acids arginine, proline, phenylalanine, leucine, tyrosine, glycine and histidine. Interestingly, roles have been reported for some of these amino acids in asthma. For instance, increased plasma concentrations of phenylalanine and histidine have been reported to be associated with adverse effects in asthma, including increased inflammation, histamine production and airway hyper-responsiveness. In addition, arginine metabolism is implicated in asthma pathogenesis, as increased levels of the arginine metabolite L-ornithine have been reported in paediatric patients, and administration of arginine attenuated inflammation in an in vivo asthma model. Based on these reports, it is possible that amino acid metabolism by MRX004 may be involved in the anti-asthma effects of this strain.

[0280] Comparative Rapid ID 32A analysis was carried out between MRX004 and four B. breve type strains, which are annotated in Figure 80B as Bif Ref 1 (DSM 20091), Bif Ref 2 (DSM 20213), Bif Ref 6 (JCM 7017) and Bif Ref 7 (UCC2003). This analysis demonstrated that MRX004 was the only strain tested to ferment the polysaccharide raffinose, which may be significant, because raffinose is involved in the production of bacterial components such as exopolysaccharides, and raffinose fermentation can also reportedly confer effects on the host such as increased caecal butyrate, increased gastrointestinal proliferation and weight loss.

[0281] API ®< 50 CH testing was carried out to further examine carbohydrate metabolism in MRX004. As per manufacturer's instructions, bacteria were cultured in 10 ml YCFA broth for 16-18 hours at 37°C in an anaerobic workstation. This culture was diluted in 10 ml API ®< CHL Medium so as to achieve a density roughly equivalent to McFarland standard No. 2, and 110 µl of this mixture was used to inoculate each cupule on a set of API ®< 50 CH test strips. Test strips were incubated in a humidified incubation box at 37 °C in an anaerobic workstation for 48 hours, following which the colour of each cupule was recorded and assigned a value of negative, intermediate positive, positive or doubtful.

[0282] Using API ®< 50, MRX004 tested positive for utilisation of the following carbohydrate sources: amidon (starch), amygdalin, arbutin, cellobiose, esculin, galactose, gentiobiose, glucose, glycogen, fructose, fucose, lactose, maltose, mannose, mannitol, melibiose, melezitose, methyl α-D-glucopyranoside, N-acetylglucosamine, ribose, saccharose (sucrose), salicin, sorbitol, trehalose, turanose and xylitol. These results correlated with those obtained for Rapid ID 32A testing in that MRX004 demonstrated fermentation of galactose, glucose, mannose and raffinose in both test systems. Interestingly, some MRX004 carbohydrate substrates, namely galactose and fructose, may be implicated in the mechanism of action of this strain, based on their reported effects in the literature. Galactose α-1,3- galactose derived from meat sources is a known allergen and causative agent of anaphylaxis, and intake levels of dietary fructose are correlated with increased asthma severity. Taken together both sets of API ®< data for MRX004 suggest that the metabolism of this strain may play a role in its anti-asthma effects.Example 14 - Genome analysis

[0283] A comparison of the genome content of strain MRX004 and the reference strains of B. breve, 1, 2, 6 and 7 was carried out using blastn as part of the BLAST+ 2.3.0 suite of programs. A maximum E-value cut-off score of 10E-5 was employed throughout the analysis.

[0284] 333 genes were identified (Table 1) that are present in the genome of strain MRX004 but are absent from the B. breve reference strains 1 (DSM 20091), 2 (DSM 20213), 6 (JCM 7017) and 7 (UCC2003). Many of the genes listed in Table 1 are frequently observed as being hypervariable among B. breve strains

[80] . As expected, the regions of variability include genes that code for proteins involved in carbohydrate metabolism and transport, phage-associated genes, mobile elements, as well as 173 genes predicted to encode proteins or genes of unknown function.

[0285] Genes that are present in MRX004 but absent from B. breve reference strains 1, 2, 6 and 7 are listed in Table 1. Genes that are not highlighted are absent in more than one of the four reference strains. The large number of genes that are present in MRX004 but are not present in numerous B. breve reference strains suggests that MRX004 is distinct from and / or distinguishable from these known B. breve strains. Genes highlighted with single underlining are present in MRX004 but absent in B. breve reference strain 1. Genes highlighted with double underlining and in bold are present in MRX004 but absent in B. breve reference strain 2. Genes highlighted with italics are present in MRX004 but absent in B. breve reference strain 6. A maximum E-value cut-off score of 10E-5 was employed for the blastn analysis. Table 1 4DBb 0021cMultiple sugar ABC transporter, substrate-binding protein4DBb_0023Probable LacI-type transcriptional regulator4DBb 0024Sucrose-6-phosphate hydrolase (EC 3.2.1.B3)4DBh 0026c Maltodextrin glucosidase (EC 3.2.1.20) 4DBb 0036cHypothetical protein4DBb 0038cMSM (multiple sugar metabolism) operon regulatory protein4DBb_0119cHypothetical protein4DBb 0120cHypothetical protein4DBb_0187Hypothetical protein4DBb_0188Hypothetical protein4DBb 0203cCell division protein FtsL4DBb_0204cHypothetical protein4DBb 0205cHypothetical protein4DBb_0206cTranscriptional regulator, HxlR family4DBb 0207Rrf2-linked NADH-flavin reductase4DBb 0208ATP-dependent DNA helicase RecG-related protein4DBb 0209cHypothetical protein4DBb_0210Putative transporter4DBb_0211Omega-3 polyunsaturated fatty acid synthase subunit, PfaA4DBb_0212Type I polyketide synthase4DBb_0213cHypothetical protein4DBb 0214cHypothetical protein4DBb 0215Hypothetical protein4DBb_0216cConserved hypothetical protein4DBb 0218cHypothetical protein4DBb_0219cDNA-cytosine methyltransferase4DBb 0220cHypothetical protein4DBb 0221cHypothetical protein4DBb 0222cHypothetical protein4DBb_0223cIntegrase4DBb_0256Hypothetical protein4DBb 0257cLacI-type transcriptional regulator4DBb_0258Putative glycosyl hydrolase of unknown function (DUF1680)4DBb_0284Transcriptional regulator, AraC family4DBb_0285N-Acetyl-D-glucosamine ABC transport system, sugar-binding protein4DBb 0286Sugar ABC transporter permease4DBb_0287N-Acetyl-D-glucosamine ABC transport system, permease protein 24DBb 0288Alpha-galactosidase (EC 3.2.1.22)4DBb_0329cATPase component BioM of energizing module of biotin ECF transporter4DBb 0330Major facilitator superfamily MFS 14DBb 0368GMP synthase [glutamine-hydrolyzing] (EC 6.3.5.2)4DBb_0369cMu-like prophage protein gp294DBh_0410 Putative galactosidase 4DBb_0419Hypothetical protein4DBb 0421Glycosyltransferase SypP4DBb 0422Capsular polysaccharide biosynthesis protein4DBb 0423Hypothetical protein4DBb 0424Glycosyltransferase4DBb_0425Membrane protein involved in the export of O-antigen, teichoic acid lipoteichoic acids4DBb 0426Glycosyltransferase (EC 2.4.1.-)4DBb_04272-succinyl-5-enolpyruvyl-6-hydroxy-3- cyclohexene-1-carboxylic-acid synthase (EC 2.2.1.9)4DBb 0428Hypothetical protein4DBb_0429Mobile element protein4DBb 0430Hypothetical protein4DBb 0431cHypothetical protein4DBb_0432cMobile element protein4DBb 0433cHypothetical protein4DBb_0434cHypothetical protein4DBb 0435Hypothetical protein4DBb 0436Hypothetical protein4DBb 0437cHypothetical protein4DBb_0438cMobile element protein4DBb_0439cHypothetical protein4DBb_0440cMobile element protein4DBb_0518cPIN domain protein4DBb_0519cHypothetical protein4DBb_0555cHypothetical protein4DBb_0556cHypothetical protein4DBb_0557cLSU ribosomal protein L31p @ LSU ribosomal protein L31p, zinc-independent4DBb_0558cSSU ribosomal protein S14p (S29e) @ SSU ribosomal protein S14p (S29e), zinc-independent4DBb_0559cLSU ribosomal protein L33p @ LSU ribosomal protein L33p, zinc-independent4DBb_0560cHypothetical protein4DBb_0561Hypothetical protein4DBh_0613c Cellulose synthase (UDP-forming) (EC 2.4.1.12) 4DBh_0614 Chitinase (EC 3.2.1.14) 4DBh_0615 Sensory box / GGDEF family protein 4DBb_0660Mobile element protein4DBb 0662Mobile element protein4DBb_0663cNeuraminidase NanP4DBb 0664Hypothetical protein4DBb 0665Hypothetical protein4DBb_0666Mobile element protein4DBb_0667Mobile element protein4DBb_0668Mobile element protein4DBb 0718Predicted biotin regulatory protein BioR (GntR family)4DBb_0719Hypothetical protein4DBb 0720Hypothetical protein4DBb_0778Hypothetical protein4DBb_0789cMobile element protein4DBb_0790cMobile element protein4DBb_0837cPossible conserved integral membrane protein.4DBb 0840Macrolide-efflux protein4DBb_0866Cation-transporting ATPase, E1-E2 family4DBb 0867Hypothetical protein4DBb_0872Hypothetical protein4DBb_0879cTranscriptional regulator, Cro / CI family4DBb 0880cIntegral membrane protein4DBb_0946MFS general substrate transporter4DBb 0947cPutative membrane protein4DBb_0948cMobile element protein4DBb_0952cHypothetical protein4DBb 0953Hypothetical protein4DBb_0954cHypothetical protein4DBb 0955cHypothetical protein4DBb 0956cPredicted permeases4DBb 0957Narrowly conserved hypothetical protein4DBb 0958cConserved hypothetical protein4DBb 0986cHvpothetical protein4DBb 0987cDuplicated ATPase component BL0693 of energizing module of predicted ECF transporter4DBb 0988cTransmembrane component BL0694 of energizing module of predicted ECF transporter4DBb_1009cPhage holin4DBb_1010cMembrane-bound lytic murein transglycosylase D precursor (EC 3.2.1.-)4DBb 1011cHypothetical protein4DBb 1012Hypothetical protein4DBb_1013cHypothetical protein4DBb_1014cHypothetical protein4DBb_1015cHypothetical protein4DBb_1016cHypothetical protein4DBb 1017cPhage tail protein4DBb_1018cPhage tail length tape-measure protein4DBb_1019cHypothetical protein4DBb_1020cHypothetical protein4DBb_1021cHypothetical protein4DBb_1022cHypothetical protein4DBb_1023chypothetical protein4DBb_1024cHypothetical protein4DBb_1025cHypothetical protein4DBb_1026cHypothetical protein4DBb_1027cHypothetical protein4DBb_1028cHypothetical protein4DBb_1029cHypothetical protein4DBb_1030cPhage protein4DBb 1031cPhage terminase, large subunit # Pham24DBb_1032cPhage terminase, large subunit4DBb_1033chypothetical protein4DBb 1034hypothetical protein4DBb 1035hypothetical protein4DBb_1036cFIG00424913: hypothetical protein4DBb_1037chypothetical protein4DBb_1038chypothetical protein4DBb_1039chypothetical protein4DBb_1040chypothetical protein4DBb_1041chypothetical protein4DBb_1042chypothetical protein4DBb_1043chypothetical protein4DBb_1044chypothetical protein4DBb_1045chypothetical protein4DBb_1046chypothetical protein4DBb 1047cChromosome (plasmid) partitioning protein ParB4DBb_1048chypothetical protein4DBb_1049chypothetical protein4DBb_1050cSingle-stranded DNA-binding protein4DBb_1051chypothetical protein4DBb_1052chypothetical protein4DBb_1053chypothetical protein4DBb_1054chypothetical protein4DBb_1055chypothetical protein4DBb_1056hypothetical protein4DBb_1057chypothetical protein4DBb_1058chypothetical protein4DBb_1059hypothetical protein4DBb_1060hypothetical protein4DBb 1061hypothetical protein4DBb_1062chypothetical protein4DBb_1063chypothetical protein4DBb 1064putative phage integrase4DBb_1113Permeases of the major facilitator superfamily4DBb 1142hypothetical protein4DBb_1143chypothetical protein4DBb 1172Integrase4DBb_1173chypothetical protein4DBb_1174cNarrowly conserved hypothetical protein4DBb 1175hypothetical protein4DBb_1176chypothetical protein4DBb 1177ABC transporter, ATP-binding protein4DBb 1178hypothetical protein4DBb_1179hypothetical protein4DBb 1180hypothetical protein4DBb_1181hypothetical protein4DBb_1182two-component system sensor kinase4DBb_1183hypothetical protein4DBb_1203cregulatory protein, LacI4DBb_1204cFIG01131316: hypothetical protein4DBb_1205ctransport system permease4DBb_1206cPredicted rhamnose oligosaccharide ABC transport system, permease component 24DBb_1207cextracellular solute-binding protein, family 14DBh_ 1212c hypothetical protein 4DBb_1213chypothetical protein4DBb_1214Mobile element protein4DBb_1215Mobile element protein4DBb_1219Hypothetical protein4DBb_1220cHypothetical protein,4DBb_1221cProtein of unknown function DUF262 family4DBb_1222cHypothetical protein4DBb_1223Hypothetical protein4DBb_1224Mobile element protein4DBb_1234cABC-type sugar transport system, periplasmic component4DBb_1235Hypothetical protein4DBb_1328cPutative phage integrase4DBb_1329Hypothetical protein4DBb_1330Hypothetical protein4DBb_1331cHypothetical protein4DBb_1332Hypothetical protein4DBb 1333Hypothetical protein4DBb_1334Hypothetical protein4DBb_1335cHypothetical protein4DBb_1336cNegative regulator of beta-lactamase expression4DBb_1337cHypothetical protein4DBb_1338cHypothetical protein4DBb_1339cHypothetical protein4DBb 1340cHypothetical protein4DBb_1341cHypothetical protein4DBb_1342cHypothetical protein4DBb_1343cHypothetical protein4DBb_1344Hypothetical protein4DBb_1345cPhage tail fiber protein4DBb_1346cHypothetical protein4DBb_1347cPhage minor tail protein4DBb_1348cHypothetical protein4DBb_1349cHypothetical protein4DBb_1350cPhage protein4DBb_1351cPhage protein4DBb_1352cPhage protein4DBb_1353cPhage protein4DBb_1354cHypothetical protein4DBb 1355cHypothetical protein4DBb_1356cPhage major capsid protein #Fam0025 # Pham1644DBb_1357cPutative phage prohead protease4DBb_1358cPhage portal protein4DBb_1359cgp2, terminase4DBb_1360cHypothetical protein4DBb_1361cHypothetical protein4DBb_1362cHypothetical protein4DBb_1363cHypothetical protein4DBb_1364cHypothetical protein4DBb_1365cHypothetical protein4DBb_1366cHypothetical protein4DBb_1367cHypothetical protein4DBb_1368cHypothetical protein4DBb_1369cHypothetical protein4DBb_1370cHypothetical protein4DBb_1371cHypothetical protein4DBb_1372cSingle-stranded DNA-binding protein4DBb_1373cHypothetical protein4DBb_1374cRecombinational DNA repair protein RecT (prophage associated)4DBb_1375cphage-related protein4DBb_1376cHypothetical protein4DBb_1377cHypothetical protein4DBb_1378cHypothetical protein4DBb_1379cHypothetical protein4DBb_1380cMethyltransferase (EC 2.1.1.-)4DBb_1381cHypothetical protein4DBb_1382cHypothetical protein4DBb_1383cHypothetical protein4DBb_1384Hypothetical protein4DBb_1385Hypothetical protein4DBb_1386Hypothetical protein4DBb_1387Hypothetical protein4DBb_1388Hypothetical protein4DBb_1456Sucrose permease, major facilitator superfamily4DBb_1486cEsterase / lipase4DBb_1487cGlucose / mannose:H+ symporter GlcP4DBb_1488cTwo-component response regulator yesN4DBb_1533 cHypothetical_protein 4DBb_1534cHypothetical protein4DBb_1535cType I restriction-modification system, restriction subunit R (EC 3.1.21.3)4DBb_1536cATP-dependent DNA helicase recG (EC 3.6.1.-)4DBb_1537cType I restriction-modification system, specificity subunit S (EC 3.1.21.3)4DBb_1538cType I restriction-modification system, DNAmethyltransferase subunit M (EC 2.1.1.72)4DBb_1539cHypothetical protein4DBb_1540Type I restriction-modification system, specificity subunit S (EC 3.1.21.3)4DBb_1541Integrase4DBb_1542cType I restriction-modification system, specificity subunit S (EC 3.1.21.3)4DBb_1545cRibose ABC transport system, high affinity permease RbsD (TC 3.A.1.2.1)4DBb_1546cRibose ABC transport system, periplasmic ribose-binding protein RbsB (TC 3.A.1.2.1)4DBb_1547cRibose ABC transport system, permease protein RbsC (TC 3.A.1.2.1)4DBb_1548cRibose ABC transport system, ATP-binding protein RbsA (TC 3.A.1.2.1)4DBb_1550cHypothetical protein4DBb_1551Hypothetical protein4DBb_1552cLacI-type transcriptional regulator4DBb_1553cSimilar to tetracycline resistance protein4DBb_1554cRibokinase (EC 2.7.1.15)4DBb_1555cNADH-dependent butanol dehydrogenase A (EC 1.1.1.-)4DBb_1556cPhosphoglycolate phosphatase (EC 3.1.3.18)4DBb_1557cInosine-uridine preferring nucleoside hydrolase (EC 3.2.2.1)4DBb_1558cFructokinase (EC 2.7.1.4)4DBb_1559cPhosphoribosylanthranilate isomerase (EC 5.3.1.24)4DBb_1560cATPase component STY3233 of energizing module of queuosine-regulated ECF transporter4DBb_1561cATPase component of general energizing module of ECF transporters4DBb_1562cTransmembrane component STY3231 of energizing module of queuosine-regulated ECF transporter4DBb_1563cSubstrate-specific component STY3230 of queuosine-regulated ECF transporter4DBb_1564Hypothetical sugar kinase in cluster with indigoidine synthase indA , PfkB family of kinases4DBb_1569Transcriptional regulator, TetR family4DBb_1570cEsterase / lipase4DBb_1571cHypothetical protein4DBb_1572cHypothetical protein4DBb_1573COG1309: Transcriptional regulator4DBb_1574cHypothetical protein4DBb_1578cMelibiose carrier protein4DBb_1579cHypothetical protein4DBb_1580cTranscriptional regulator, TetR family4DBb_1581Hypothetical protein4DBb_1582Two-component response regulator colocalized with HrtAB transporter4DBb_1583Hypothetical protein4DBb_1584cHypothetical protein4DBb_1585Sensor histidine kinase4DBb_1586Mg(2+) transport ATPase protein C4DBb_1587Transcriptional regulator, AbrB family4DBb_1588Hypothetical protein4DBb_1620c3'-to-5'oligoribonuclease (orn)4DBb_1769cTranscriptional activator MltR4DBb_1770cXylitol dehydrogenase (EC 1.1.1.9)4DBb_1771cRibitol / Xylitol / Arabitol transporter, MFS superfamily4DBb_1773cGlyoxalase family protein4DBb_1774cRibitol / Xylitol / Arabitol transporter, MFS superfamily4DBb_1775cSorbitol dehydrogenase (EC 1.1.1.14)4DBb_1926cMembrane protein, related to Actinobacillus protein (1944168)4DBb_1928cGlycosyl transferase, group 2 family protein4DBb_1929cTeichoic acid export ATP-binding protein TagH (EC 3.6.3.40)4DBb_1930cRhamnose-containing polysacharide translocation permease4DBb_1934cHypothetical protein4DBb_1935Possible glycosyltransferase4DBb_1936cCell wall surface anchor family protein4DBb_1937cD-alanyl-D-alanine carboxypeptidase (EC 3.4.16.4)4DBb_1965Hypothetical protein4DBb_2010cMobile element protein4DBb_2011Transcriptional regulator, LacI family4DBb_2012Xylose ABC transporter, periplasmic xylose-binding protein XylF4DBb_2013Ribose ABC transport system, ATP-binding protein RbsA (TC 3.A.1.2.1)4DBb_2014Ribose ABC transport system, permease protein RbsC (TC 3.A.1.2.1)4DBb_2015Hypothetical protein4DBb_2016Mobile element protein4DBb_2028cBeta-glucosidase (EC 3.2.1.21) Sequences

[0286] SEQ ID NO:1 (consensus 16S rRNA sequence for strain 751) SEQ ID NO:2 (strain 751 genome sequence) - see electronic sequence listing of PCT / GB2016 / 051776. REFERENCES

[0287] [1] Spor et al. (2011) Nat Rev Microbiol. 9(4):279-90. [2] Eckburg et al. (2005) Science. 10;308(5728):1635-8. [3] Macpherson et al. (2001) Microbes Infect. 3(12):1021-35 [4] Macpherson et al. (2002) Cell Mol Life Sci. 59(12):2088-96. [5] Mazmanian et al. (2005) Cell 15;122(1):107-18. [6] Frank et al. (2007) PNAS 104(34):13780-5. [7] Scanlan et al. (2006) J Clin Microbiol. 44(11):3980-8. [8] Kang et al. (2010) Inflamm Bowel Dis. 16(12):2034-42. [9] Machiels et al. (2013) Gut. 63(8):1275-83.

[10] WO 2013 / 050792

[11] WO 03 / 046580

[12] WO 2013 / 008039

[13] WO 2014 / 167338

[14] Goldin and Gorbach (2008) Clin Infect Dis. 46 Suppl 2:S96-100.

[15] Azad et al. (2013) BMJ. 347:f6471.

[17] Srůtková et al. (2011) J. Microbiol. Methods, 87(1):10-6.

[18] Ye et al. (2015) PLoS One. 10(1):e0117704.

[19] Fabro et al. (2015) Immunobiology. 220(1):124-35.

[20] Yin et al. (2014) Immunogenetics. 66(3):215-8.

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[22] Schieck et al. (2014) J Allergy Clin Immunol. 133(3):888-91.

[23] Balato et al. (2014) J Eur Acad Dermatol Venereol. 28(8):1016-24.

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[25] Fahy (2009) Proc Am Thorac Soc 6.256-259

[51] Miyamoto-Shinohara et al. (2008) J. Gen. Appl. Microbiol., 54, 9-24.

[52] Cryopreservation and Freeze-Drying Protocols, ed. by Day and McLellan, Humana Press.

[53] Leslie et al. (1995) Appl. Environ. Microbiol. 61, 3592-3597.

[54] Mitropoulou et al. (2013) J Nutr Metab. (2013) 716861.

[55] Kailasapathy et al. (2002) Curr Issues Intest Microbiol. 3(2):39-48.

[56] Handbook of Pharmaceutical Excipients, 2nd Edition, (1994), Edited by A Wade and PJ Weller

[57] Remington's Pharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985)

[58] Handbook of MicrobiologicalMedia, Fourth Edition (2010) Ronald Atlas, CRC Press.

[59] Maintaining Cultures for Biotechnology and Industry (1996) Jennie C. Hunter-Cevera, Academic Press

[60] Strobel (2009) Methods Mol Biol. 581:247-61.

[61] Gennaro (2000) Remington: The Science and Practice of Pharmacy. 20th edition, ISBN: 0683306472.

[62] Molecular Biology Techniques: An Intensive Laboratory Course, (Ream et al., eds., 1998, Academic Press).

[63] Methods In Enzymology (S. Colowick and N. Kaplan, eds., Academic Press, Inc.)

[64] Handbook of Experimental Immunology, Vols. I-IV (D.M. Weir and C.C. Blackwell, eds, 1986, Blackwell Scientific Publications)

[65] Sambrook et al. (2001) Molecular Cloning: A Laboratory Manual, 3rd edition (Cold Spring Harbor Laboratory Press).

[66] Handbook of Surface and Colloidal Chemistry (Birdi, K.S. ed., CRC Press, 1997)

[67] Ausubel et al. (eds) (2002) Short protocols in molecular biology, 5th edition (Current Protocols).

[68] PCR (Introduction to Biotechniques Series), 2nd ed. (Newton & Graham eds., 1997, Springer Verlag)

[69] Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987) Supplement 30

[70] Smith & Waterman (1981) Adv. Appl. Math. 2: 482-489.

[71] Brand et al. (2007) Nature Protocols. 2(5):1269-1275

[72] Jiao et al. (2014) Immunopathology and Infectious Diseases. 184(4):1085-93.

[73] Rockwell et al., (1972) J Natl Cancer Inst. 49:735-49.

[74] Bertram and Janik (1980) Cancer Lett. 11:63-73.

[75] Darlington (1987) Meth Enzymol. 151:19-38.

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[80] Bottacini et al (2014) BMC Genomics. 15:170, DOI: 10.1186 / 1471-2164-15-170, PMID: 24581150 SEQUENCE LISTING <110> 4D PHARMA RESEARCH LIMITED <120> COMPOSITIONS COMPRISING BACTERIAL STRAINS <130> P075775EP <150> GB 1510467.2 <151> 2015-06-15 <150> GB 1520501.6 <151> 2015-11-20 <160> 1 <170> SeqWin2010, version 1.0 <210> 1 <211> 1439 <212> DNA <213> Unknown <220> <223> Strain 751 consensus 16S rRNA sequence <400> 1

Claims

1. A composition comprising a bacterial strain having a genome with at least 90% sequence identity to SEQ ID NO: 2 for use in a method of treating or preventing allergic asthma, eosinophilic asthma or airway hyper-responsiveness associated with asthma.

2. The composition for use according to claim 1, wherein the bacterial strain has a genome with at least 92%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 2 across 100% of said SEQ ID NO:2.

3. The composition for use according to any preceding claim, wherein the bacterial strain utilises the following carbohydrate sources: amidon, amygdalin, arbutin, cellobiose, esculin, galactose, gentiobiose, glucose, glycogen, fructose, fucose, lactose, maltose, mannose, mannitol, melibiose, melezitose, methyl α-D-glucopyranoside, N-acetylglucosamine, ribose, saccharose, salicin, sorbitol, trehalose, turanose and xylitol.

4. The composition for use according to any preceding claim, wherein the bacterial strain provides the same pattern as the bacterium deposited under accession number NCIMB 42380 when analysed by amplified ribosomal DNA restriction analysis when using a Sau3AI restriction enzyme.

5. The composition for use according to any preceding claim, wherein the bacterial strain has low adherence to Caco-2 cells in YCFA compared to Bifidobacterium breve.

6. The composition for use according to any preceding claim, wherein the bacterial strain produces more bound surface exopolysaccharides compared to Bifidobacterium breve.

7. The composition for use according to any preceding claim, wherein the bacterial strain has a 16s rRNA sequence that is at least 99%, 99.5% or 99.9% identical to SEQ ID NO:1, or wherein the bacterial strain has the 16s rRNA sequence represented by SEQ ID NO: 1.

8. The composition for use according to any preceding claim, wherein the bacterial strain comprises at least 5, 10, 20, 50 or all of the genes in Table 1.

9. The composition for use according to any preceding claim, wherein the bacterial strain is deposited under accession number NCIMB 42380.

10. The composition for use according to any preceding claim, for use in a method of reducing an eosinophilic inflammatory response in the treatment or prevention of the asthma.

11. The composition for use according to any preceding claim, for use in reducing peribronchiolar infiltration in the treatment of the allergic asthma.

12. The composition for use according to any preceding claim, for use in a patient with elevated eosinophil levels, as identified through blood sampling or sputum analysis.

13. The composition for use according to any preceding claim, for use in a method of reducing IL-17 production or reducing Th17 cell differentiation in the treatment or prevention of the asthma; and / or wherein the composition is for use in a patient with elevated IL-17 levels or Th17 cells.

14. The composition for use according to any preceding claim, wherein the composition is formulated as a probiotic, and is not combined with a prebiotic compound.

15. The composition for use according to any preceding claim, wherein the composition is for oral administration, and / or wherein the composition comprises one or more pharmaceutically acceptable excipients or carriers, and / or wherein the bacterium is lyophilised.