Compositions comprising branched (1,3)(1,6)-beta-glucans for use in the treatment of cancer

Abstract

The present invention relates to a physiologically acceptable composition for use in the treatment of a cancer, wherein the physiologically acceptable composition comprises a first branched (1,3)(1,6)-β-glucan and a second branched (1,3)(1,6)-β-glucan different from the first. In an embodiment, the treatment comprises training the immune system, in particular the innate immune system.

Description

[0001] P134821PC00 Title: Physiologically acceptable compositions comprising β-glucans for use in the treatment of cancer. The invention relates to a physiologically acceptable composition, comprising β-glucans, for use in the treatment of a cancer. In particular, a composition for use according to the invention is suitable to induce training of the innate immunity and can be used for the treatment of a person having cancer or a person having a genetic predisposition relating to cancer. Traditionally, the vertebrate immune system has been categorized as falling into two different categories, namely the innate immune system and adaptive immune system. The innate immunity recognizes pathogens and tissue damage in a non-specific way through pattern recognition receptors (PRRs), which sense pathogen-associated and damage-associated molecular patterns. The innate immune system is usually very effective in eliminating invading pathogens and is very rapid. On the other hand, the adaptive immune system responses depend on interaction between dendritic cells (DCs) that present antigens and specialized T cells and B cells. The lymphocyte-dependent adaptive immune responses are slower to develop, but are antigen specific and lead to long-term immunological memory. Recently, the general understanding of innate immunity has been changed. Research has indicated that also the innate immune system shows adaptive characteristics and can present some memory feature of a first encounter, a concept termed ‘trained innate immunity’ or ‘trained immunity’ (Netea et al., 2011. Cell Host Microbe 9:355-361). This concept describes the long-term functional reprogramming of innate immune cells, which is evoked by exogenous or endogenous insults and which leads to an altered response towards a second challenge after the return to a non-activated state (Netea et al., 2020. Nat Rev Immunol 20: 375-388). The secondary response to the subsequent non-specific stimulus can be altered in such a way that the cells respond more potently or less strongly than to the primary response, conferring context-adjusted and time- adjusted responses. For certain infections and vaccinations, a broad protection against the same and unrelated pathogens through innate immune mechanisms has been observed. For example, in mouse models the BCG vaccine has been shown to be protective against candidiasis. Similarly in humans, live vaccines such as the BCG vaccine, measles vaccine, smallpox vaccine and oral polio vaccine, have been shown to provide non-specific protective effects against infections other than the target disease (Benn et al., 2013. Trends Immunol 34: 431-439). Another example is a prior mild infection with C. albicans that confers protection against the subsequent exposure to a normally lethal candidiasis infection in mice (Bistoni et al., 1986. Infect Immun 51: 668-674). Thus, the innate immune system has the ability to generate memory responses after infections or vaccinations to mount rapid and enhanced immune responses upon exposure to secondary infections. Vaccination with the BCG vaccine has been shown to induce anti-tumour immune effects leading to the prevention or treatment of malignancies such as bladder cancer, melanoma, leukaemia and lymphoma. This anti-cancer effect of the BCG vaccine seemed to be dependent on the induction of trained innate immunity in monocytes and macrophages (Buffen et al., 2014. PLoS Pathogen 10:e1004485). However, the use of BCG vaccine has many disadvantages. One of the disadvantages is that the vaccine is difficult to manufacture leading to chronic shortages. Furthermore, mould contamination has been reported to be occurring often during the manufacturing process. Another disadvantage of the BCG vaccine is that it comprises live bacteria, which can induce systemic infection that can be life-threatening in patients with a defect in host defence, such as immunocompromised patients. The inventors consider that promoting trained immunity could be beneficial in different therapeutic contexts such as the treatment and prevention of a cancer. β-Glucans are part of the cell wall of fungi and yeasts and have been studied intensively for over 50 years. They are inter alia known to have immunomodulating effects boosting immunity against cancer, see Anticancer Research March 2018, 38 (3) 1327-1333. During the last few years, it was shown that β-glucan is also a strong inducer of trained immunity. Human monocytes that are pre-exposed to β-glucan exhibit an enhanced immune response upon re- stimulation with various microbial ligands in vitro (Quintin et al., 2012. Cell Host Microbe 12: 223-232). Furthermore, in vivo animal studies demonstrated that treatment with β-glucan offers macrophage-mediated protection from subsequent challenge with pathogens, including Candida albicans and Staphylococcus aureus (Bistoni et al., 1986. Infect Immun 51: 668-674; Quintin et al., 2012. Cell Host Microbe 12: 223-232). Considering the short lifespan of myeloid cells in the circulation, the mechanism responsible for the long-lasting protective effects of β- glucan was initially unclear. However, a recent study by Mitroulis et al. (2018. Cell 172:147-161) revealed that β-glucan not only induces trained immunity in mature monocytes and macrophages, but that it also alters the functional program of hematopoietic progenitors in the bone marrow, which likely accounts for the prolonged generation of trained myeloid cells in the circulation. Similar adaptations at the level of the bone marrow have been observed for other inducers of trained immunity such as BCG vaccine (Kaufmann et al., 2018. Cell 172: 902- 904) and a high-fat diet (Christ et al., 2018. Cell 172:162-175). Recently, β-glucan- mediated trained immunity was suggested as an effective and novel treatment option for pancreatic cancer, wherein the treatment effect was observed both when a yeast-derived β-glucan was administered intraperitoneally (IP) and orally (Woeste et al., 2023. Journal for ImmunoTherapy of Cancer 11:e006221). There is a continuing need to provide alternative products suitable for use in the treatment or prevention of a cancer. It is in particular an object to provide a composition for use in the treatment of a cancer, which is suitable to provide an improved activation of the innate immunity compared to a known composition, e.g. the BCG vaccine or a known composition comprising a β-glucan from for example Candida albicans. One or more further objects that may be addressed will follow from the description herein below. We have now found that one or more of said objects are realized by providing a specific composition comprising at least two differently branched (1,3)(1,6)β- glucans. Accordingly, the invention relates to a physiologically acceptable composition for use in the treatment of a cancer, wherein said physiologically acceptable composition comprises a first branched (1,3)(1,6)-β-glucan and a second branched (1,3)(1,6)-β-glucan different from the first. As illustrated in the Examples, below, a composition for use according to the invention, comprising at least two different types of branched (1,3)(1,6)β-glucans (as active ingredients), has been found effective in training of the innate immune system. More specifically, a combination of two different branched (1,3)(1,6)β- glucans lead to an improved effect compared to the individual effects of the glucans. In particular, a strong amplification of the production of proinflammatory cytokine TNF-α and a moderate increase of the production of proinflammatory cytokine IL-6 was observed by induction using a combination (referred to as ABB i16), which is a combination of two different branched (1,3)(1,6)β-glucans, being S. cerevisiae β-glucans ABB i24 and ABB i25. The observed increase in production of proinflammatory cytokines was higher for the combination ABB i16 compared to the individual effects of ABB i24 and ABB i25, as well as compared to the effect induced by a Candida albicans β-glucan known to be able to induce trained immunity (Quintin et al., 2012. Cell Host Microbe 12: 223-232). It was furthermore identified that a complexity of multiple receptors and intracellular signalling pathways play a role in the observed induction of the trained innate immunity by the combination of both branched (1,3)(1,6)β-glucans. Further, administration of the β-glucan combination ABB i16 also conferred protection against the development of cancer in multiple mice cancer models, including a bladder cancer model and melanoma model. Thus, the use in the treatment of a cancer by therapy in accordance with the invention can either be the treatment of a cancer that has manifested in a human or animal having a cancer or be a prophylactic treatment, which prophylactic treatment can reduce the risk of developing a cancer. Furthermore, it is shown in the examples, below, that a composition according to the invention, comprising at least two different branched (1,3)(1,6)β- glucans can modulate the balance between various T helper (Th) lymphocyte responses. In particular it is shown that using a combination of two different branched (1,3)(1,6)β-glucans (referred to as ABB i16), a heightened Th1 cell response is induced. Furthermore, an elevated secretion of the Th2 immunomodulatory cytokine IL-10 and a decrease in IL-5 / IL-10 ratio was observed by ABB i16 administration. Furthermore, ABB i16 was shown to have a role in the induction of a regulatory Th17 response, characterized by increased production of IL-22 and at the same time no induced production of IL-17, which is the prototype pro-inflammatory product of Th17 responses. IL-17 is known to be associated with autoimmune disease phenotypes, suggesting that it can be safely used as an oral supplement for patients with autoimmune conditions. The involvement of ABB i16 in the different Th responses, shows that ABB i16 has the potential to regulate the Th1 / Th2 / Th17 balance towards antimicrobial and regulatory effects. Interestingly, the effects of a combination of two different branched (1,3)(1,6)β-glucans could be enhanced by co-administration with other compounds. In particular, co-stimulation of ABB i16 alongside Bacille Calmette-Guerin (BCG), a combination of IL-1β and IL-23 or a combination of IL-12 and IL-18 was found to further enhance Th1 polarization and IL-22 induction, hinting at an additive effect of ABB i16 on both Th1 and regulatory Th17 immune responses. Figure 1: Schematic representation of the structure of a branched (1,3)(1,6)-β- glucan. This figure is adapted from Waszkiewicz-Robak (“Spent Brewer’s Yeast and Beta-Glucans Isolated from Them as Diet Components Modifying Blood Lipid Metabolism Disturbed by an Atherogenic Diet”. Lipid Metabolism, edited by Rodrigo Baez, IntechOpen, 2013. 10.5772 / 51530). Figure 2: Gas chromatography (GC) chromatogram of the linkage residues of different tested β-glucan comprising compositions. Glycosyl linkage analysis performed by combined gas chromatography / mass spectrometry (GC-MS) of the partially methylated alditol acetate (PMAA) derivatives produced by hydrolysis of the β-glucan compositions. Chromatogram of’, β-glucan ABB i25 and β-glucan ABB i24 are shown in panels (A) and (B) respectively. Figure 3: Trained immunity induction by various β-glucans. Monocytes were trained with 1, 2, 5 and 10 µg / ml of Candida albicans (Ca) β-glucan or Saccharomyces cerevisiae (Sc) β-glucans for 24 hours. On the 6 day after resting, macrophages were re-stimulated with 10 ng / ml of LPS and incubated for 24 hours. TNF-α and IL-6 production was measured in supernatants by ELISA (n=6 for both cytokines) at day 7. Data in bar plots are represented as mean ± SEM. Statistical analysis was performed by Mann-Whitney U test. *P < 0.0332, **P < 0.0021; differs from Ca β-glucan control, or #p < 0.0332 differs between Sc β-glucans 1,2 and 3- stimulated conditions. Tested β-glucans include Sc β-glucan ABB i24, Sc β-glucan ABB i25 and a combination of β-glucans ABB i24 and ABB i25, which is indicated as “Sc β-glucan ABB i16”. Figure 4: Receptors and signalling pathways involved in β-glucan trained immunity induction. Monocytes were pre-incubated for 1h with RPMI, IgG2B Mouse, or IgG2b Goat (positive controls), anti-hDectin-1 antibody, anti-CR3 antibody, and TLR-4 inhibitor Bart.-LPS (A), or DMSO (positive control), Syk kinase inhibitor, Raf-1 inhibitor GW5074, and PI 3-kinase inhibitor SL-2052 (B), or with RPMI, IgG2B Mouse or IgG2b Goat (positive controls), anti-hDectin-1 antibody, anti-MMR, anti-CR3 antibody, and TLR-4 inhibitor Bart.-LPS (C) then primed for 24 hr with Candida albicans (Ca) β-glucan or Saccharomyces cerevisiae (Sc) β-glucan ABB i16 (i.e. a combination of glucans ABB i24 and ABB i25). Subsequently washed, rested for 5 days, and at day 6 restimulated for 24h with 10 ng / ml LPS. TNF-α and IL-6 production was measured in supernatants by ELISA, n=6. Data in bar plots are represented as mean ± SEM. Statistical analysis was performed by Mann-Whitney U test. *p < 0.05, **p < 0.01 compared to control for panels (A) and (B); and *p < 0.05 compared to control for panel (C). Figure 5: Tumor weight reduction in a murine model of melanoma (A) and bladder cancer (B) after induction of the innate immunity. Mice were inoculated with either B16 / F10 melanoma cells or MB49 bladder cancer cells and tumor weight (in mg) was measured after 14-days. The effects of a PBS injection (placebo) or ABB i16 (1 mg administered i.p.) was compared. Figure 6. Tumor volume and tumor weight reduction in a murine model of melanoma. Mice were inoculated with B16 / F10 melanoma cells cancer cells and tumor volume (in mm3) was measured daily between day 9 and 14 after tumor cell inoculation (A). The effects of a PBS injection (“WT CONTROL”) or ABB i16 (1 mg administered i.p., “WT BGLUCAN”) was compared. On day 14, the tumor weight in mg was compared between control and ABB i16-treated mice (B). Figure 7: Dose-response analysis of cytokine production after PBMC stimulation with different β-glucan concentrations.(A) schematic timeline of in vitro direct stimulation experiments. (B) Th1, (C) Th2 and D) Th17 cytokine levels after stimulation of human PBMCs for 7 days in the presence of HPS with RPMI and ABBi16 (1, 10, and 100 µg / mL) (n=6). Data are presented as mean ± SEM. Figure 8: Comparison of Th1 / Th2 / Th17 cytokine profiles induced through PBMC stimulation with ABB i16 and C. albicans. (A) schematic timeline of in vitro direct stimulation experiments. (B) Th1, (C) Th2 and (D) Th17 cytokine levels after stimulation of human PBMCs for 7 days in the presence of HPS with either RPMI, ABBi16 (10 µg / mL) or heat-killed C. albicans (1 × 106microorganisms / mL), as indicated (n=6). Data are presented as mean ± SEM. Figure 9: Additive effects of ABB i16 on BCG-induced Th1 / Th2 / Th17-specific cytokine production. (A) schematic timeline of in vitro direct stimulation experiments. (B) Th1, (C) Th2 and (D) Th17 cytokine levels were assessed after stimulation of human PBMCs for 7 days in the presence of HPS with either RPMI, ABB i16 (10 µg / mL), BCG (5 µg / mL), or BCG combined with ABB i16, as indicated (n=6). Data are presented as mean ± SEM. Figure 10: ABB i16 enhances the synergistic effect of IL-1β+IL-23 and IL- 12+IL-18 on Th cell cytokine production. (A) schematic timeline of in vitro direct stimulation experiments. (B) IFN-γ, TNF, and IL-22 concentrations were assessed after stimulation of human PBMCs for 7 days in the presence of HPS with either RPMI, ABB i16 (10 µg / mL), IL-1 in combination with IL-23 (both 100 ng / mL), or IL-1β and IL-23 together with ABB i16, as indicated (n=6). (C) Th1, Th2 and Th17 cytokine concentrations after stimulation of human PBMCs for 7 days in the presence of HPS with either RPMI, ABB i16 (10 µg / mL), IL-12 in combination with IL-18 (both 10 ng / mL), or IL-12 and IL-18 together with ABBi16, as indicated (n=6). Data are presented as mean ± SEM. Figure 11: Cytokine-driven Th1 / Th2 / Th17 polarization in the presence of ABBi16. (A) IL-5, IL-10, and IL-17 concentrations were assessed after stimulation of human PBMCs for 7 days in the presence of HPS with either RPMI, ABBi16 (10 µg / mL), IL-1 in combination with IL-23 (both 100 ng / mL), or IL-1 and IL-23 together with ABBi16, as indicated (n=6). (B) IL-5, IL-10, and IL-17 levels after stimulation of human PBMCs for 7 days in the presence of HPS with either RPMI, ABBi16 (10 µg / mL), IL-12 in combination with IL-18 (both 10 ng / mL), or IL-12 and IL-18 together with ABBi16, as indicated (n=6). Data are presented as mean ± SEM. For the purpose of clarity and a concise description, features are described herein as part of the same or separate embodiments, however, it will be appreciated that the scope of the invention may include embodiments having combinations of all or some of the features described. Definitions As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well. The term "or" includes any and all combinations of one or more of the associated listed items, unless the context clearly indicates otherwise (e.g. if an “either ….or” construction is used). It will be understood that the terms "comprises" and "comprising" specify the presence of stated features but do not preclude the presence or addition of one or more other features. It will be further understood that when a particular step of a method is referred to as subsequent to another step, it can directly follow said other step or one or more intermediate steps may be carried out before carrying out the particular step, unless specified otherwise. In the context of this application, the term "about" means generally a deviation of 15 % or less from the given value, in particular a deviation of 10% or less, more in particular a deviation of 5% or less. As is used herein, the term “physiologically acceptable composition”, refers to a composition that is suitable for administration to an individual such as an animal or a human. As is known in the art, the term “β-glucan”, refers to a polysaccharide at least composed of a chain (i.e. backbone) of a plurality of glucopyranose (i.e. D-Glucose) molecules linked by β-glycosidic bonds. Optionally, the β-glucan may further comprise one or more saccharide units other than glucopyranose, such as mannose units or N-acetylglucosamine units, e.g. at least 0.5 %, at least 1 % or at least 2 % of the number of monosaccharide units in the β-glucan. The number of monosaccharide units other than glucopyranose is smaller than the number of glucopyranose, typically up to 40 %, up to 30 %, up to 20 % up to 10 % or up to 5 % of the number of monosaccharide units in the β-glucan. Some β-glucans are continual repeats of monosaccharide units (i.e. unbranched), while other β-glucans may be branched and comprise different kinds of β-glycosidic bonds. In particular, β-glucans useful in accordance with the present invention are β-glucans obtained from or obtainable from yeast cell walls having predominately β-(1,3) and β-(1,6) glycosidic bonds. The designation β-(1,3) glycosidic bond for a glycosidic linkage indicates that the etheric oxygen bridge between two consecutive monosaccharide units of the β- glucan connects the number 1 carbon of the first unit to the number 3 carbon of the second unit, and that the etheric oxygen bridge attaches to carbon 1 of the first unit from above the ring. The designation β-(1,6) glycosidic bond for a glycosidic linkage indicates that the etheric oxygen bridge between two consecutive monosaccharide units of the β- glucan connects the number 1 carbon of the first unit to the number 6 carbon of the second unit, and that the etheric oxygen bridge attaches to carbon 1 of the first unit from above the ring. As is used herein, the term “(1,3)(1,6)-β-glucan” or “β-1,3 / 1,6-glucan”, refers to a β-glucan having a backbone (chain) of monosaccharide units, in particular glucopyranose units, linked by β-1,3 glycosidic bonds and further one or more monosaccharide units linked via a β-1,6 glycosidic bond. The branches in a (1,3)(1,6)-β-glucan are usually from about 1 to 6 monosaccharide units long. As is used herein, the term “a different β-glucan”, in particular refers to a β- glucan that differs in terms of structure, in particular in terms of the sequence of different monosaccharides, in terms of monosaccharide composition or in terms of difference in branches, compared to a β-glucan it is compared to. A person skilled in the art will be able to determine if two β-glucans are different from each other based on scientific literature, for example based on Boutros et al. (2022. Food Chemistry 367:130708). Boutros et al. describe the variation between different β- glucan lots of one (1,3)(1,6)-β-glucan source as well as the variation between (1,3)(1,6)-β-glucans from a different source. It was found that for the same beta- glucan, the measured molar percentages for different linkages differ with maximum a few percentage points over the lots, see Table 2 of Boutros et al.. Boutros et al. also describe the variation between beta-glucans from a different source, see e.g. Table 3 of Boutros et al., and observed that this variation was larger than the variation between β-glucans from the same source. It was further found that every proprietary source of yeast β-glucan that was tested had a unique purity profile, branching, and linkage pattern that determines the chemical structure and composition. To determine if two or more β-glucans are different or not, a person skilled in the art can determine the purity profile, branching and / or linkage pattern of the β-glucans and determine if the variation observed falls outside the ranges of variation between β-glucans from a same source (see Table 2 of Boutros et al.) and / or if the variation observed falls within the ranges of variation between β-glucans from a different source (see Table 3 of Boutros et al.) described by Boutros et al.. As is used herein, the term “individual”, refers to any living organism such as an animal or human that can benefit from the administration of a physiologically accepted composition for use according to the invention. The term “animal” as used herein, refers in particular to vertebrate animals including fish, birds, mammals, reptiles and amphibians. The animal can be a farm animal, domestic animal or laboratory animal. An individual, which may be treated in accordance with the invention may in particular be selected from humans, nonhuman primates and monkey species, cows, sheep, pigs, goats, horses, dogs, cats, rodents such as mice, rats and guinea pigs, poultry, such as chickens, hens, turkeys, ducks and geese and aquatic animals such as fish and shrimp. The term individual does not denote a particular age or sex (such as male / female). Thus, humans of any age group, including adults (older than 18 years of age) and children (0-18 years of age), e.g. newborn individuals (0-12 months of age), toddlers (12-36 months of age), can be treated in accordance with the invention. In a preferred embodiment, the composition is for use in the treatment of a human. As is used herein, the term “nutritional product”, refers to a composition intended for ingestion by an individual providing at least one nutrient to the individual. Nutritional products generally comprise one or more components selected from the group consisting of protein, fat, carbohydrate and micro- nutrients. As is used herein, the term “nutraceutical”, refers to any nutritional product providing an extra health benefit in addition to the basic nutritional value found in foods. As is used herein, the term “cosmeceutical”, refers to any cosmetic product with bioactive ingredients purported to have a health benefit. As is used herein, the term “trained immunity” or “trained innate immunity”, refers to the long-term functional reprogramming of innate immune cells, which is evoked by exogenous or endogenous insults and which leads to an altered response towards a second challenge after the return to a non-activated state (Netea et al., 2020. Nature Reviews Immunology 20: 375-388). As is used herein, the term “inactivated”, or “dead”, or “non-viable”, refers to an organism, such as a yeast, not being capable of reproduction or colonization. An inactivated organism can have intact or broken cell membranes. The skilled person will be able to obtain an inactivated organism yeast based on common general knowledge and the information disclosed herein. Possible means include irradiation, heat inactivation, sonication, lyophilization and chemical inactivation. As used herein, the term “heat-killed”, refers to an organism inactivated by heat-treatment and such not capable of metabolic activity nor colonization. Means of heat-treatment to inactivate an organism are known to a person skilled in the art and include tyndallization, pasteurization, ultra-high temperature (UHT) heating, Ohmic heating (or Joule heating), blanching, drying, boiling and sterilization. As is used herein, the term “tyndallization”, refers to a sterilization process often used to heat-kill probiotic microorganisms. Tyndallization involves repeating a heat-killing step for a certain consecutive days, for example as described on page 11 in the Handbook of Microbiological Media (third edition, 2004, CRC Press) by Ronald M. Atlas. Therefore, the term “tyndallized”, refers to an organism having been heat-killed by tyndallization and such not capable of metabolic activity or colonization. As is used herein, the term “alive”, or “viable”, refers to an organism capable of reproduction or colonization. As used herein, the term “cell lysis” refers to any type of cell disruption that results in the release of intercellular biological components naturally contained in the cells of an organism. Therefore, the term “lysate”, refers to a product obtained after cell lysis. A “lysate”, as used herein, means in particular essentially the entire lysate obtained by lysis of an organism and therefore comprises macromolecules such as DNA, RNA, proteins, peptides and lipids from the lysed cell; as well as cellular debris, including cell wall material, and cell membrane components from the lysed cells. Methods for obtaining a lysate are known to a person skilled in the art and include enzymatic, physical and chemical methods. Cell wall components can be separated from the fluid part of the lysate e.g. by centrifugation of filtering. As used herein, the term “extract” of a microorganism, refers to a fluid part or fraction of the microorganism cell or lysate, in particular liquid contents of yeast or bacterial cells, in particular obtainable by filtration or by centrifugation, or a fraction thereof, obtainable by extraction from the cells or the lysate, using an extracting phase. As used herein, the term “cancer’, refers to a group of diseases that can start in almost any organ or tissue of the body when abnormal cells grow uncontrollably, go beyond their usual boundaries to invade adjoining parts of the body and / or spread to other organs (as defined by World Health Organisation, WHO). The latter process is called metastasizing and is a major cause of death from cancer. A neoplasm and malignant tumour are other common names for cancer. Cancers are mainly classified in two ways: by the type of tissue in which the cancer originates (histological type) and by primary site, i.e. the location in the body where the cancer first developed (topography). A person skilled in the art can consult e.g. International Classification of Diseases for Oncology, 3rd Edition (ICD-O-3) for definitions regarding and classification of cancer-types. According to ICD-O-3, there are six major categories of histological cancer types, being carcinoma, sarcoma, myeloma, leukaemia, lymphoma and mixed types. Beta-glucans β-glucans form a natural component of the cell walls of bacteria, fungi, yeast, and cereals such as oat and barley. Each type of β-glucan comprises a different molecular backbone, level of branching, and molecular weight which affects its solubility and physiological impact. Β-glucans found in the cell walls of yeast, such as Saccharomyces cerevisiae, contain a 1,3-carbon backbone with elongated 1,6 carbon branches. In yeast, β-glucans act as a barrier with the main role in supporting the cell wall due to a rigid structure. Yeast cell wall ranges between 15– 30 wt% of the dry mass of the yeast that has in its composition 50 wt% β-1,3- glucans and 10 wt% β-1,6-glucans, meaning that yeast S. cerevisiae is theoretically composed of a mixed-linkage of β-1,3 / 1,6-glucans between 9 wt% and 18 wt% (Avramia et al.,2021. Int J Mol Sci 22: 825). Methods to extract β-glucans from different sources such as yeast are known to the person skilled in the art and include for example extraction with hot alkali and acetic acid followed by aqueous washes to remove soluble components in the cell walls. A more detailed protocol is for example described by EP 0466037 B1. Alternatively, a β-glucan can be obtained by synthesis. It is known in the art that complex branched glucans can be obtained by synthesis, see e.g. US2718179 and WO2013176542. Βeta-glucans act by interacting with different receptors on white blood cells in the immune system such as macrophages, neutrophils, granulocytes, natural killer cells, dendritic cells, and corresponding cells found in tissue surfaces. These receptors include the Toll-like receptor (TLR) family but also non-TLR receptors such as dectin-1 receptor or complement receptor 3 (CR3). It was found that dectin- 1 is a type II C-type lectin receptor that specifically recognize (1,3)- and / or (1,6)- linked β-glucans, and that recognition of β-glucan by dectin-1 induces several dectin-1-mediated cellular responses (Brown, 2005. Nature Review Immunology 6: 33-43). These cellular responses include, the production of pro-inflammatory cytokines and chemokines, such as tumour-necrosis factor (TNF) and CXC- chemokine ligand 2 (CXCL2), interleukin-12 (IL-12) and interferon-γ (IFN-γ). When a TLR receptor and dectin-1 receptor interact simultaneously, they are reported to complement each other and produce a different and more desirable response than when the receptors are independently activated (Yadav & Schorey, 2006. Blood 108: 3168-3175; Ferwerda et al., 2008. Cell Microbiol 10: 2058-2066; Castro et al., 2020. Mol Nutr Food Res 65: e190107). It is furthermore known that the ability of β-1,3 / 1,6-glucans to activate innate immune cells depends on its branched structure: e.g. it was found that a β-1,3- glucan without any side chains (branches) does not activate macrophages. (Raa, 2015. Special Issue: Gut in Focus 26: 27824). Similarly, products with only one single glucose molecule in the ‘side chain’, as in most mushroom beta-1,3 / 1,6- glucans, were found to have lower macrophage activating activity than yeast cell- wall beta-1,3 / 1,6-glucan (Raa, 2015. Special Issue: Gut in Focus 26: 27824). In addition to chain length, the frequency of side chains was shown essential for immune-stimulating and immune-modulating ability of beta-1,3 / 1,6-glucans (Bohn & BeMiller, 1995. Carbohydrate Polymers 28: 3-14). These molecular structures may change or be destroyed during extraction, hence affecting the biological activity of the extracted product. The physiologically acceptable composition for use according to the invention comprises a first branched (1,3)(1,6)-β-glucan and a second branched (1,3)(1,6)-β- glucan different from the first branched (1,3)(1,6)-β-glucan. Without being bound to theory, the inventors discovered that a stimulation of the immune system with different (1,3)(1,6)-β-glucans having a different degree of branching, allows the correct spatial disposition to stimulate multiple different β-glucan-receptors and multiple intracellular pathways involved in the trained innate immunity induction simultaneously, leading to the observed improved (synergistic) effect on trained immunity, as shown in the examples below (Figure 4). The first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β-glucan of the composition for use according to the invention preferably are recognised by (can bind to) at least two receptors chosen from the group consisting of dectin-1, TLR-4, and CR3, more preferably the first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β-glucan activate dectin-1, TLR-4 and CR3. The activation of a certain receptor (due to the binding) can for example be measured by using receptor inhibition experiments, e.g. as generally known in the art or as used in the Examples (for dectin-1, TLR-4 and CR3). Alternatively or in addition, the first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β-glucan of the composition according to the invention preferably are activated by at least two receptors chosen from the group consisting of dectin-1, TLR-4, MMR and CR3, preferably the first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β-glucan activate at least three receptors chosen from the group consisting of dectin-1, TLR-4, MMR and CR3, more preferably the first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β-glucan activate dectin-1, TLR-4, MMR and CR3. Preferably, the physiologically acceptable composition for use according to the invention further comprises a mannan. A mannan is a polysaccharide at least composed of a chain (i.e. backbone) of a plurality of mannose molecules linked by β- glycosidic bonds. Mannans are known to increase pro-inflammatory cytokine production in a TLR4 dependent manner (Tada et al., 2002. Microbiology and Immunology 46: 503-512). Therefore, the presence of mannan in the physiologically acceptable composition for use according to the invention can contribute to stimulation of receptors and intracellular pathways involved in the trained innate immunity induction, providing a stronger induction of trained immunity. If the physiologically acceptable composition comprises mannan, the content of the mannan is preferably 0.5-10 wt% based on the total weight of the composition, more preferably the content of the mannan is 0.5-6.0 wt% based on the total weight of the composition, most preferably the content of the mannan is 0.5-4.5 wt% based on the total weight of the composition. Said first branched (1,3)(1,6)-β-glucan preferably is free of 3-linked mannose units and / or free of 4,6 linked glucose units and / or free of 2,3,4 linked glucose residues. The first branched (1,3)(1,6)-β-glucan has preferably 0 – 2 %, preferably 0 – 0.6 %, preferably 0.1 – 0.5 %, more preferably 0.2-0.4 %, such as about 0.3 %, terminally linked mannose units of the total number of monosaccharide units in the β-glucan. The first branched (1,3)(1,6)-β-glucan has preferably 5-13%, preferably 6-12%, preferably 7-11%, more preferably 8-10%, such as about 8.7 % terminally linked glucose units of the total number of monosaccharide units in the β-glucan. The first branched (1,3)(1,6)-β-glucan has preferably 45-80%, preferably 50-75%, preferably 55-70%, more preferably 60-66%, such as about 63 % 1,3 linked glucose units of the total number of monosaccharide units in the β-glucan. The first branched (1,3)(1,6)-β-glucan has preferably 7-16%, preferably 8-15%, preferably 9- 14%, more preferably 10-13%, such as about 11.4 %, 1,6 linked glucose units of the total number of monosaccharide units in the β-glucan. The first branched (1,3)(1,6)- β-glucan has preferably 0-5%, preferably 0.5-3.5%, preferably 0.5-2.5%, more preferably 1-2 %, such as about 1.5 %, 1,4 linked glucose units of the total number of monosaccharide units in the β-glucan. The first branched (1,3)(1,6)-β-glucan has 0-5%, preferably 0.5-2.5%, preferably 0.5-1.5%, more preferably 0.75-1.25 %, such as about 1 %, 3,4 linked glucose units of the total number of monosaccharide units in the β-glucan. The first branched (1,3)(1,6)-β-glucan has 1-10%, preferably 2-9%, preferably 3-8%, preferably 4-7%, more preferably 5-6 %, such as about 5.4 % , 2,3 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 5-13 %, preferably 6-12 %, preferably 7-11 %, more preferably 8-10 %, such as about 8.8%, 3,6 linked glucose units of the total number of monosaccharide units in the β-glucan. Most preferably the first branched (1,3)(1,6)-β-glucan is ‘Auxoferm Glucan Max’ as commercially available (Ohly GmbH), which is also named ABB i25. Details of the glucan structure, such as glycosyl linkages, can be analysed in a manner known per se, e.g. by gas chromatography / mass spectrometry (GC-MS) of partially methylated alditol acetate (PMAA) derivatives produced from the samples containing the glucans, based on the procedure described by Willis et al. (2013) PNAS, 110 (19) 7868-7873. Said first branched (1,3)(1,6)-β-glucan preferably has a lower terminally linked mannose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a higher terminally linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)- β-glucan preferably has a lower 1,3 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a lower 1,3 linked mannose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a higher 1,6 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)- β-glucan preferably has a higher 1,4 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a lower 3,4 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a lower 2,3 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a higher 3,6 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a lower 4,6 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a lower 2,3,4 linked glucose content than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a side chain length of 1.5-3.5, preferably 2-3, more preferable 2.2-2.4, such as about 2.3, most preferably 2.3. The second branched (1,3)(1,6)-β-glucan preferably has a side chain length of 1.5-3.5, preferably 2-3, more preferable 2.4-2.6, such as about 2.5, most preferably 2.5. The first branched (1,3)(1,6)-β-glucan preferably has a lower side chain length than the second branched (1,3)(1,6)-β-glucan. The first branched (1,3)(1,6)-β-glucan preferably has a side chain frequency of 5-11, preferably 6-10, more preferable 7-9, such as about 8, most preferably 8. The second branched (1,3)(1,6)-β-glucan preferably has a side chain frequency of 8-14, preferably 9-13, more preferable 10-12, such as about 11, most preferably 11. The first branched (1,3)(1,6)-β-glucan preferably has a lower side chain frequency than the second branched (1,3)(1,6)-β-glucan. Said second branched (1,3)(1,6)-β-glucan has preferably 0-3%, more preferably 0-1.5%, preferably 0.2-1%, more preferably 0.4-0.8 %, such as about 0.6 %, terminally linked mannose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 1-9%, preferably 2-8%, preferably 3-7%, preferably 4-6%, more preferably 4.5-5.5 %, such as about 5.1 %, terminally linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 45-85%, preferably 50-80%, preferably 55-75%, preferably 60-70%, more preferably 62-68 %, such as about 65.2 %, 1,3 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β- glucan has preferably 0-5%, preferably 0.5-3%, preferably 0.75-2%, preferably 1- 1.8%, more preferably 1.2-1.6 %, such as about 1.4%, 1,3 linked mannose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 4-14%, preferably 5-13%, preferably 6-12%, preferably 7-11%, preferably 8.5-10.5%, more preferably 9-10 %, such as about 9.5 %, 1,6 linked glucose units of the total number of monosaccharide units in the β- glucan. The second branched (1,3)(1,6)-β-glucan has preferably 0-5%, preferably 0.25-3%, preferably 0.5-2%, more preferably 1-1.7 %, such as about 1.3 %, 1,4 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 0-5%, preferably 0.5-4%, preferably 1-3%, more preferably 1.2-2 %, such as about 1.6 %, 3,4 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 4-15%, preferably 5-12%, preferably 6- 11%, preferably 7-10%, more preferably 7.5-9 %, such as about 8.2 %, 2,3 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 2-10%, preferably 3-9%, preferably 4-8%, preferably 5-7%, more preferably 6-6.6 %, such as about 6.3 %, 3,6 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 0-5%, preferably 0.2-2%, preferably 0.4-1.5%, more preferably 0.6-1 %, such as about 0.8 %, 4,6 linked glucose units of the total number of monosaccharide units in the β-glucan. The second branched (1,3)(1,6)-β-glucan has preferably 0-6%, preferably 1-5%, preferably 2-4%, more preferably 2.5-3.5 %, such as about 3.1 %, 2,3,4 linked glucose units of the total number of monosaccharide units in the β-glucan. Most preferably the second branched (1,3)(1,6)-β-glucan is ‘Auxoferm HGT’ as commercially available (Ohly GmbH), which is also named ABB i24. In the physiologically acceptable composition for use according to the invention, preferably at least one branched (1,3)(1,6)-β-glucan is composed of glucose units and one or more further monosaccharide units different from a glucose unit, which further monosaccharide unit(s) form up to 3 % of the monosaccharide units of said branched (1,3)(1,6)-β-glucan, preferably 1 – 3 % of the monosaccharide units of said branched (1,3)(1,6)-β-glucan, most preferably about 2 % of the monosaccharide units of said branched (1,3)(1,6)-β-glucan. Preferably said glucan composed of glucose units and one or more further monosaccharide units is a mannoglucan. Advantageously, the glucose content of the mannoglucan is 95.5- 99.5% and the mannose content is 0.5 – 4.5 %, preferably the glucose content is 97 - 99.5% and the mannose content is 0.5 - 3 %, more preferably the glucose content is 97.5 - 98.5% and the mannose content is 1.5 - 2.5 %; herein the glucose content is calculated as the number of glucose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan. The mannose content is calculated as the number of mannose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan. Preferably the first branched (1,3)(1,6)-β-glucan has a lower mannose content than the second branched (1,3)(1,6)-β-glucan, more preferably the mannose content of the first glucan being in the range of 0 - 1.5% and the relative mannose content of the second glucan preferably being more than 1.5%, in particular up to 6 %, more in particular up to 3 %; herein the mannose content is calculated as the number of mannose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan. A physiologically acceptable composition for use according to the invention comprises an amount of the at least two (1,3)(1,6)-β-glucans together of at least 10 wt%, preferably an amount of at least 20%, more preferably an amount of at least 30%, more preferably an amount of at least 40%, , more preferably an amount of at least 50 wt%, more preferably an amount of at least 60 wt%, more preferably an amount of at least 70 wt%, more preferably an amount of at least 80 wt%, more preferably an amount of at least 90 wt%, more preferably an amount of at least 95 wt%, such as an amount of about 99 wt% or about 100 wt% or an amount of 100 wt%, based on the total weight of the composition. A physiologically acceptable composition for use according to the invention preferably comprises an amount of the at least two (1,3)(1,6)-β-glucans together of at least 10 wt%, preferably an amount of at least 20%, more preferably an amount of at least 30%, more preferably an amount of at least 40%, more preferably an amount of at least 50 wt%, more preferably an amount of at least 60 wt%, more preferably an amount of at least 70 wt%, more preferably an amount of at least 80 wt%, more preferably an amount of at least 90 wt%, more preferably an amount of at least 95 wt%, such as an amount of about 99 wt% or about 100 wt% or an amount of 100 wt%, based on the dry weight of the composition. A physiologically acceptable composition for use according to the invention may comprises an amount of the at least two (1,3)(1,6)-β-glucans together of at least between 40-90 wt% based on the total weight of the composition or based on dry weight of the composition. Preferably, the physiologically acceptable composition for use according to the invention comprises a weight ratio of the first (1,3)(1,6)-β-glucan to the second (1,3)(1,6)-β-glucan between 0.5 and 15 preferably between 1 and 12, more preferably between 2 and 11, more preferably between 3 and 10, in particular a weight ratio in the range of 7 to 9. Other components of a physiologically acceptable composition In addition to (1,3)(1,6)-β-glucans, a physiologically acceptable composition for use according to the invention may also comprise at least one microorganism or a component (other than a (1,3)(1,6)-β-glucan), a lysate or an extract thereof. Preferably, said microorganism is a yeast. A preferred yeast included in the physiologically acceptable composition for use according to the invention is a Saccharomyces cerevisiae. Said yeast may be live or inactivated. Preferably, the inactivated yeast is heat-killed. Advantageously, the yeast is mineral-enriched. Preferably, the yeast is rich in zinc, selenium or both. Further components that may be present in the physiologically acceptable composition include protein, fat, ash, additives, excipients and other carbohydrates such as mannose and a mannan. Furthermore, the physiologically acceptable composition for use according to the invention advantageously includes BCG and / or a combination of cytokines. Preferably the combination of cytokines is a combination of IL-β1 with IL-23 or a combination of IL-12 and IL-18. Administration mode and formulations of a physiologically acceptable composition The physiologically acceptable composition for use according to the present invention may be administered systemically. Advantageously, the composition is administered topically, in particular directly to a specific site where cancer cells or precancerous cells are present or to a specific site where an increased risk of the development of cancer cells or precancerous cells is expected (such as due to a genetic disposition or exposure to a potentially carcinogenic event). Thus, in a specific embodiment, the composition comprising the beta-glucans is for use in the treatment of a person having a genetic predisposition relating to cancer to prevent development of a malignity. Administration can be achieved in any way effective for delivering the composition to cancer cells or precancerous cells, which administration may be based on a known administration mode for a specific cancer to be treated. For example, administration methods include oral, topical, intradermal, transdermal, parenteral, subcutaneous, intramuscular, intraperitoneal, intrabursal, intratumoral, or intranasal instillation, intracavitary or intracapsular instillation, intraocular, intraarterial, intralesional, or by application to mucosa such as the mucosa of the nose, throat and bronchi. The composition is in particular suitable for oral, subcutaneous, intradermal, intra- tumoral or local administration. The physiologically acceptable composition for use according to the present invention may be administered into the gastrointestinal tract, e.g. orally. In a specific embodiment, the composition is administered by tube feeding or as a suppository. Formulations of a composition for use according to the invention suitable for oral intake include but are not limited to: capsules, coated capsules, tablets, sachets, pills, pearls, soft-gels, vials, powders, granules, solutions, suspensions, emulsions, elixirs, syrups, sprays, lozenges, troches, gums, hard candies and gels. Formulations of a composition for use according to the invention suitable for topical administration include compositions suitable for application to a part of the gastro-intestinal tract at which the effect of the yeast components is needed, such as a suppository or a gastro-intestinal medical device. Administration into the gastrointestinal tract may be topically. Topical administration into the gastrointestinal tract generally comprises administration at mucosa or epithelium of the gastro-intestinal tract. A medical product comprising the composition for use according to the invention can be a product suitable to create a protective biofilm or the like at the surface of the gut epithelium. In a specific embodiment, the composition is to be administered as a sustained release product. The composition for use according to the invention may be a food product. The food product may be a fermented food product or a non- fermented food product. Examples of a particularly suitable food products include dairy products such as a yogurt, a yogurt drink, cheese, milk, milk powder, infant formula, cream, ice-cream, cream powder and butter; fruit-based products such as fruit juice, compote or fruit jelly; solid foodstuffs such as flours, cereals, a snack, a biscuit, and liquid formulations such as vegetable beverages, smoothies, isotonic drinks, salts solutions and enteral nutrition recipes. The physiologically acceptable composition for use according to the invention for intake by an animal may be any appropriate food product for animals and include, in addition to the food products listed above, tablets, coated tablets, granules, cereal grains, (dried) meat, (dried) fish, oil meals, cakes, cookies, sugarcane, and roughages such as grasses, hays, silage, root crops, straw and stover. The nutritional product for use according to the invention may be in the form of a dietary supplement, a food additive, a feed additive, a functional food in human nutrition, a functional food in animal nutrition, a food additive or functional ingredient for nutraceuticals or food for special medical purposes. The physiologically acceptable composition for use according to the invention can be a pharmaceutical product or a nutraceutical product. Said pharmaceutical product may further comprise a pharmaceutically acceptable adjuvant and / or excipient. Adjuvants and excipients are well known to a person skilled in the art. A pharmaceutical product in accordance with the present invention may include one or more further active ingredients. Some examples of active ingredients of which one or more may advantageously be included in a pharmaceutical product in accordance with the present invention are corticosteroids (like prednisone and budesonide); calcineurin inhibitors (like cyclosporine and tacrolimus); biologics (like adalimumab and infliximab); allergy medications like antihistamines (like cetirizine and loratadine), corticosteroids (like fluticasone), and biologics for allergies (like dupilumab and omalizumab); immune modulators (like methotrexate and azathioprine). It is also possible to use a pharmaceutical product according to the present invention with another treatment, like immunoglobulin therapy or recombinant human interleukin-7. A nutraceutical product according to the present invention may further comprise one or more nutrients, in particular one or more nutrients selected from proteins, digestible carbohydrates, lipids, vitamins and trace elements. Examples of preferred ingredients of which one or more can be included in nutraceutical products in accordance with the present invention are omega-3 fatty acids, curcumin and probiotics. They may help modulate immune responses. The physiologically accepted composition may further comprise a bulking agent, in particular a carbohydrate, for example maltodextrin. The production of a physiologically acceptable composition comprising β- glucans The physiologically acceptable composition for use according to the invention can be produced by blending the different components. Generally, a method for producing a composition for use in accordance with the invention comprises providing the first branched (1,3)(1,6)-β-glucan from a first source, in particular originating from a first organism, providing the second branched (1,3)(1,6)-β-glucan from a second source different from the first source, in particular originating from a different organism and blending the first branched (1,3)(1,6)-β-glucan and the second first branched (1,3)(1,6)-β-glucan. The (1,3)(1,6)-β-glucans may be provided as at least substantially pure materials or as part of a composition containing the respective branched β-glucan and one or more other ingredients. Optionally, one or more other components may be included in the composition, e.g. components for the composition mentioned elsewhere herein. In particular, one or more further components may be added selected from the group of additives, excipients, protein, ash, micro-organisms, (further) micro-organism components, cell-lysates and cell- extracts. Methods of treating an individual and training an individual’s innate immune system In accordance with the invention, the physiologically acceptable composition is advantageously for use in the treatment of an individual with a cancer. In a preferred embodiment, the use comprises the training, in particular the induction of training, of the innate immunity of said individual. In a further preferred embodiment, the use in the treatment of an individual with a cancer comprises the reduction in volume, weight or both of a malignant tumour of said individual. In a further preferred embodiment, the use in the treatment of an individual with a cancer comprises providing an increased life-expectancy or prognosis of survival. In a further preferred embodiment, the treatment of an individual with a cancer comprises improvement of the immune system, in particular an improvement in the innate immune system. In a further embodiment, the improvement of the immune system comprises treatment of an imbalanced T helper (Th) cell response such as an Th1 / Th2 response and / or imbalanced Th1 / Th2 / Th17 response. This is also advantageous in the treatment of a cancer patient. For instance, Th1 cells produce IFN-γ and IL-12. These have anticancer activity. Th2 cells mediate the humoral immune. They produce IL-4 and IL-10. IL-4 can inhibit the secretion of IFN-γ. Thus, a disbalanced Th1 / Th2 response (too high Th2 response) and / or imbalanced Th1 / Th2 / Th17 response can actually promote cancer procession. A cancer to be treated according to the invention can be any histological type of cancer. The cancer to be treated can be a carcinoma e.g. an adenocarcinoma or squamous cell carcinoma; a sarcoma e.g. osteosarcoma or osteogenic sarcoma (bone), chondrosarcoma (cartilage), leiomyosarcoma (smooth muscle), rhabdomyosarcoma (skeletal muscle), mesothelial sarcoma or mesothelioma (membranous lining of body cavities), fibrosarcoma (fibrous tissue), angiosarcoma or hemangioendothelioma (blood vessels), liposarcoma (adipose tissue), glioma or astrocytoma (neurogenic connective tissue found in the brain), myxosarcoma (primitive embryonic connective tissue) and mesenchymous or mixed mesodermal tumor (mixed connective tissue types); a myeloma; leukaemia e.g. myelogenous or granulocytic leukemia (malignancy of the myeloid and granulocytic white blood cell series), lymphatic, lymphocytic, or lymphoblastic leukaemia (m alignancy of the lymphoid and lymphocytic blood cell series) and polycythemia vera or erythremia (malignancy of various blood cell products, but with red cells predominating); lyphoma, e.g. Hodgkin lymphoma and non-Hodgkin lymphoma; and / or a mixed type cancer e.g. adenosquamous carcinoma, mixed mesodermal tumour, carcinosarcoma and teratocarcinoma. In a preferred embodiment, the cancer is selected from the group consisting of bladder cancer; skin cancers; osteosarcoma; colorectal cancer; lung cancer; breast cancer; and mouth and throat cancer. In a particularly preferred embodiment the cancer is selected from bladder cancer; melanoma (a skin cancer); and osteosarcoma. A cancer to be treated according to the invention can be any topological type of cancer, in particular a cancer selected from the group of bladder cancer, bone cancer (such as osteosarcoma), brain cancer, breast cancer, cervical cancer, colorectal cancer (which may be colon cancer, rectal cancer or cancer in both colon and rectum), eye cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, ovarian cancer, pancreas cancer, prostate cancer, skin cancer (such as melanoma), testicular cancer, thyroid cancer, uterine cancer, vaginal cancer, vulvar cancer, mouth cancer and throat cancer. In a preferred embodiment, the cancer is selected from the group consisting of bladder cancer, skin cancer, osteosarcoma, lung cancer, breast cancer, colorectal cancer and prostate cancer. In particular, good results have been achieved with a physiologically acceptable composition for use according to the invention, wherein the cancer is bladder cancer and melanoma. The physiologically acceptable composition for use according to the invention may be administered as a single dosage for a complete treatment or a plurality of times, dependent on the application. If multiple dosages are intended, the number of dosages is generally 10 times per day or less, in particular 1-5 times per day, although the interval between two dosages can also be more than a day, e.g. 2, 3 or 7 days. The physiologically acceptable composition may comprise or can be co- administered with a probiotic, a prebiotic, a postbiotic, an antibiotic, an analgesic, an anti-inflammatory agent, an anti-diarrhoeal agent, a vaccine such as the Bacillus Calmette-Guerin (BCG) vaccine (also referred to as BCG), an anti-cancer agent, a combination of cytokines or a mixture thereof. In particular, an increased survival rate is observed when a physiological composition for use according to the invention further comprises the BCG vaccine). The administration dosage, duration and frequency can be chosen within wide limits, dependent on the intended purpose and the subject to whom the composition is to be administered. The duration of treatment can be a relatively short period, e.g. of a week or less, or a day or less, e.g. for an acute manifestation of a disorder or symptom of a disorder. The duration of a treatment can also be prolonged, e.g. for a week or more, a month or more or a year or more, e.g. in case of a chronic disorder or in case of preventive treatment or dietary supplementation. A person skilled in the art will be able to determine an appropriate dose of the physiologically acceptable composition to administer to an individual based on common general knowledge and the information disclosed herein without undue experimentation. As the skilled person will understand, an actual preferred dosage depends on a variety of factors, including the age, body weight, general health, sex and species of the individual diet, mode and time of administration, rate of excretion, drug combination, the severity of the particular disorder, and the individual. The dosages disclosed herein are indicative of an average case for a human individual (including adults and children). There can of course be individual instances where higher or lower dosage ranges are merited. The usual effective daily dose for oral administration or other administration into the gastrointestinal tract, in particular for humans, is from about 100 mg (of the beta- glucan components) to about 1000 mg (of the beta-glucan components), preferably, from about 200 mg (of beta-glucan components) to about 900 mg (of the beta-glucan components), more preferably, from about 200 mg (of the beta-glucan components) to about 650 mg (of beta-glucan components). The usual effective daily dose for topical administration, in particular for humans, is from about 1 mg (of beta-glucan components) to about 5000 mg (of the beta-glucan components), preferably from about 5 mg (of the beta-glucan components) to about 2500 mg (of the beta-glucan components), more preferably about 25 mg (of the beta-glucan components) to about 1000 mg of the beta-glucan components), in particular from about 50 mg to about 500 mg (of the beta-glucan components). The invention provides a method for treating an individual, preferably a human, having a cancer, the method comprising an effective amount of the physiologically acceptable composition for use in accordance with the invention. The invention provides a method of treating an individual having cancer or having a genetic disposition to developing a cancer, comprising the administration of an effective amount of the physiologically acceptable composition according to the invention, hereby training the individual’s innate immune system. The invention has in particular found to be effective in increasing TNF-α. The invention provides a method of treating a subject having cancer or preventing the formation or development of a cancer in an individual in need thereof, comprising the administration of an effective amount of the physiologically acceptable composition for use according to the invention. As illustrated in the examples, a composition according to the invention has been found effective in improving TNF-α production. Therefore, the results are supporting that the composition of the invention is effective for the prevention of disorders such as a cancer associated with a low TNF-α production. Further, the examples support effective administration prior to the occurrence of a tumour to have a size-reducing effect on a tumour. Further, the effectivity of a prophylactic treatment can routinely be demonstrated, e.g. by monitoring the incidence of a disorder becoming manifesting in a cohort study wherein the incidence in a group of subjects treated with a composition according to the invention is compared with the incidence in a reference group of subjects (e.g. treated with placebo). The effect of a treatment of an individual having the disorder can be a complete cure, an alleviation of a symptom, reduced suffering etc. The invention provides a method of treating an individual with a cancer characterized by a deficient capacity to synthesize TNF-α and / or an impaired activation of immune cells such as macrophages, T lymphocytes and natural killer (NK) cells, comprising the administration of an effective amount of the physiologically acceptable composition for use according to the invention to said individual. The invention provides the use of the physiologically acceptable composition according the invention for use in the preparation of a product, which may be a medicament or a food product (such as a medical or clinical food), for use in the treatment of a cancer. The invention provides the use of the physiologically acceptable composition according the invention for the preparation of a product to train the innate immune system. The invention will now be illustrated by the following examples, which are provided by way of illustration and it will be understood that many variations in the methods described and the amounts indicated can be made without departing from the spirit of the invention and the scope of the appended claims.

[0002] EXAMPLES Example 1: Structure of different β-glucans. Materials and methods Glycosyl linkage analysis was performed by combined gas chromatography / mass spectrometry (GC-MS) of the partially methylated alditol acetate (PMAA) derivatives produced form β-glucans. The tested β-glucans are ABB i24, being ‘Auxoferm HGT’ (lot number EH22000389, obtained from Ohly GmbH, which is presently also available as ‘Auxoferm Glucan’) and ABB i25, being ‘Auxoferm Glucan Max’ (lot number EH21000534, derived from Ohly GmbH), for which the product details are shown in Table 1 and Table 2 respectively. The analysis procedure is based on the one described by Willis et al. (2013, PNAS, 110:7868-7873). Briefly, permethylation of the samples was effected by two rounds of treatment with sodium hydroxide (15 min) and methyl iodide (30 min). The samples were then hydrolyzed using 2 M TFA (2 h in sealed tube at 120 °C), reduced with NaBD4, and acetylated using acetic anhydride / TFA. The resulting PMAAs were analyzed on an Agilent 7890A GC interfaced to a 5975C MSD (mass selective detector, electron impact ionization mode); separation was performed on a 30 m Supelco SP-2331 bonded phase fused silica capillary column. Table 1: Product details of β-glucan ABB i25, being ‘Auxoferm Glucan Max’, based on product sheet dated 11 / 2021 made available by Ohly GmbH. Description AUXOFERM® Glucan is the highly concentrated cell wall fraction gained from pure primary grown yeast of the species Saccharomyces cerevisiae. The crude yeast cell walls are subjected to a gentle purification process and then finally dried. AUXOFERM® Glucan Max stands out for its content of higher molecular and non- digestible polysaccharides - especially beta glucan. The ß-(1,3) / (1,6)-linkage of glucose units in the glucan chain does allow high water binding and emulsifying capacity in different food systems. ß-(1,3) / (1,6)-glucans are furthermore known to strengthen the immune system (Vetvicka V, Vannucci L, Sima P, Richter J. (2019) Beta Glucan: Supplement or Drug? From Laboratory to Clinical Trials. Molecules. 24(7):1251. doi:10.3390 / molecules24071251). Average analysis Dry matter 96 % Protein (Nx6.25) 4 % (1,3)(1,6)-ß-D-Glucan (on dry 72 % matter basis) Ash 3 % Table 2: Product details of β-glucan ABB i24, being ‘Auxoferm HGT’, based on product sheet dated 12 / 2014 made available by Ohly GmbH. Description Auxoferm HGT is the highly concentrated cell wall fraction gained from pure culture yeast of the species Saccharomyces cerevisiae. The crude yeast cell walls are subjected to a complex purification process and then finally dried. Auxoferm HGT stands out for its content of higher molecular and indigestible polysaccharides especially glucans. The ß- (1,3) / (1,6)-linkage of glucose units in the glucan chain do allow high water binding and emulsifying capacity in different food systems. Average analysis Dry matter 96.0 % Protein ≤ 8.0 % (Nx6.25) Carbohydrates 60.0 % (total) As ß-Glucans 45.0 % Ash 10.0 % pH (in 2% 6.5 – 7.5 solution) Selected minerals Sodium < 5.00 % Potassium 0.27 % Magnesium 0.02 % Calcium 0.05 % Phosphorus 1.20 % Results Analysis of the peak areas derived from the glycosyl linkage analysis (Figure 2) is shown in Table 3. The amount of 3,6-linked glucose in ABB i25 and ABB i24 was 8.8 and 6.3% respectively. ABB i25 is shown to have a 1-6 branch of about 2.3 residues every 8thbranch of the backbone. ABB i24 is shown to have a 1-6 branch of about 2.5 residues every 11thbranch of the backbone. In contrast to ABB i24, ABB i25is not having 3-linked mannose residues, (4,6)-linked glucose residues and 2,3,4 linked glucose residues.

[0003] Table 3: Ratio of the peak areas of the glycosyl linkage analyses performed on different β-glucans. ABB i25 ABB i24 Terminal Mannopyranosyl residue (t-Man) 0.3 0.6 Terminal Glucopyranosyl residue (t-Glc) 8.7 5.1 3 linked Glucopyranosyl residue (3-Glc) 63.0 65.2 3 linked Mannopyranosyl residue (3-Man) - 1.4 6 linked Glucopyranosyl residue (6-Glc) 11.4 9.5 4 linked Glucopyranosyl residue (4-Glc) 1.5 1.3 3,4 linked Glucopyranosyl residue (3,4-Glc) 1.0 1.6 2,3 linked Glucopyranosyl residue (2,3-Glc) 5.4 8.2 3,6 linked Glucopyranosyl residue (3,6-Glc) 8.8 6.3 4,6 linked Glucopyranosyl residue (4,6-Glc) - 0.8 2,3,4 linked Glucopyranosyl residue (2,3,4-Glc) - 3.1 Side Chain Length 2.3 2.5 Side Chain Frequency n* 8 11

[0004] Example 2: Effect of β-glucans on trained immunity in innate immune cells isolated from healthy volunteers. Materials and methods Innate immunity training was tested in vitro using adherent monocytes obtained from subsequent isolation of peripheral blood mononuclear cells (PBMCs) and monocytes. The induction of trained immunity and the associated functional reprogramming of monocytes is detailed here using different β-glucans. The in vitro protocol of monocyte training was described before. For complete details on the use and execution of this protocol, reference is made to Dominguez- Andres et al. (Dominguez-Andrés et al., 2021. STAR Protocols 2, 100365). Β-glucans used in the experiments include β-1,3-(D)-Glucan, herein referred to as ‘Candida Albicans (Ca) β-glucan’, provided by David Williams (College of Medicine, Johnson City, USA) and the branched (1,3)(1,6)-β-glucans ABB i24 and ABB i25 as used in example 1. Escherichia coli lipopolysaccharide (LPS; serotype O55:B5; Sigma-Aldrich, St. Louis, MO, USA) was further purified as described previously (Hirschfeld et al., 2000. J Immunol 165:618–622). Buffy coats from healthy donors were obtained. Isolation of peripheral blood mononuclear cells (PBMCs) was performed by dilution of blood in sterile phosphate-buffered saline (PBS) and density centrifugation over Ficoll-Paque (GE Healthcare, UK). Cells were washed three times in cold PBS. Percoll isolation of monocytes was performed as described previously (Repnik et al., 2003. J Immunol Methods 278:283-292). Briefly, PBMCs were layered on top of a hyperosmotic Percoll solution (48.5% Percoll [Sigma-Aldrich, St. Louis, MO, USA], 41.5% sterile H2O, 0.16 M filter-sterilized NaCl) and centrifuged for 15 min at 580 × g. The interphase layer was isolated, and the cells were washed with cold PBS. Cells were resuspended in Dutch modified RPMI culture medium (Invitrogen, CA, USA) supplemented with 10 μg / ml gentamicin, 10 mM GlutaMAX, and 10 mM pyruvate and counted. An extra purification step was added by allowing Percoll-isolated monocytes to adhere to polystyrene flat-bottom plates (Corning, NY, USA) for 1 h at 37°C; the cells were washed with warm PBS to obtain maximal purity. PBMCs were resuspended in culture medium and the concentration was adjusted to 5 x 106cells / ml (or 500,000 cells in 100 µl / well) and added to flat-bottom 96-well plates. After incubation for 1 h at 37°C and washing with warm PBS, monocytes were incubated with culture medium only as a negative control or 1, 2, 5 and 10 μg / ml β-glucan for 24 h. To eliminate the possibility of lipopolysaccharide (LPS) contamination, all the fractions of β-glucans were pre-incubated for 1 h with and without 2 µl / ml of polymyxin B (Bedford Laboratories, Bedford, OH). After the incubation, the cells were washed once with warm PBS to remove the stimulus and allowed to rest in medium for 5 days. Subsequently, the cells were restimulated with 10 ng / ml LPS for 24 h. Hereafter, supernatants were collected and stored at −20°C until further experiments. For receptor and intracellular signalling pathways inhibition experiments, 5 × 106monocytes / ml (or 5 × 105cells in 100 μl / well) were added to flat-bottom 96- well cell culture plates and pre-incubated for 1h with antibodies and small molecule inhibitors. PRRs were blocked using 10 μg / ml of anti-Dectin-1, 10 uM of anti-CR3 antibodies, along with their respective isotype control antibodies Mouse IgG and Goat IgG (R&D Systems), and 20 ng / ml Bartonella quintana LPS (Bart. LPS, TLR4 antagonist). For the inhibition of downstream signaling mediators, monocytes were pre-incubated with 50 nM of spleen tyrosine kinase (Syk) inhibitor (574711, Sigma), 1 µM of noncanonical serine-threonine kinase Raf-1 inhibitor (GW5704, Sigma), and 10 µM of phosphoinositide-3 kinase (PI3K) inhibitor Wortmannin (SL-2052, R&D Systems). Cells were then stimulated at 37°C, 5% CO2 with 2 μg / mL β-1,3-(D)-Glucan (i.e. Ca β-glucan) and 2 μg / ml β-glucan ABB i16, i.e. a combination of ABB i24 and ABB i25 at a ratio of 12.5% of ABB i24 to 87.5% of ABB i25. The culture contained the inhibitors both during blocking (1h) and stimulation periods (24h). Supernatants were collected in the first 24 hours of stimulation to measure the production of pro-inflammatory cytokines. Afterwards, cells were washed with warm PBS, then allowed to rest in medium (with 10% human pooled serum) for 5 days. On the 6th day, the cells were restimulated with Escherichia coli LPS (10 ng / ml, serotype 055:B5, Sigma-Aldrich) for 24h, and supernatants were collected for cytokine production measurements. Cytokine production in supernatants was determined using commercial enzyme-linked immunosorbent assay kits for tumour necrosis factor alpha (TNF-α; R&D Systems, MN, USA) and IL-6 (Sanquin, Amsterdam, The Netherlands) following the instructions of the manufacturers. Differences between experimental conditions were analyzed using Mann- Whitney U test. A two-sided P value below 0.05 was considered statistically significant. All data were analyzed using GraphPad Prism software version 5.0 (La Jolla, CA, USA). Data are shown as means ± standard errors of the means. Results Results are shown in Figure 3 (A and B). Training of monocytes with the combination ABB i16 of the branched (1,3)(1,6)-β-glucans, i.e. a combination of ABB i24 and ABB i25, blended at a ratio of 12.5% of ABB i24 to 87.5% of ABB i25, induced the strongest increase in cytokine production, namely TNF-α production, upon restimulation (Figure 3), whereas the effects of ‘ABB i24’ and ‘ABB i25’ individually were less pronounced (Figure 3). The β-glucan positive control used in the experiments is a Ca β-glucan for which abilities to induce trained immunity have been well-established (Quintin et al., 2012. Cell Host Microbe 12: 223-232). Interestingly, a combination of the branched (1,3)(1,6)-β-glucans ABB i24 and ABB i25 (also referred to as combination Abb i16) was found to be significantly different in terms of induced TNF-α production from the β-glucan control (i.e. ‘Ca β-glucan’), while the branched (1,3)(1,6)-β-glucans individually (i.e. ABB i24 and ABB i25 individually) were not found to be significantly different from the β-glucan control (see Figure 3B). Additionally, Figure 3B shows a combination of the branched (1,3)(1,6)-β-glucans provided significantly different in terms of induced TNF-α production from individually administered ABB i24 and ABB i25. A situation with strong TNF-α production and moderate IL-6 release is biologically preferable over a situation with lower TNF-α but strong IL-6. The reason for this is that IL-6 induces acute phase proteins and fever, often resulting in hyperinflammation. For instance, it is involved in cytokine storm and considered a biomarker of poor prognosis. TNF-α is very relevant for host defence, released by macrophages, T-lymphocytes and Natural killer (NK) cells in response to lipopolysaccharide, other bacterial products, and interleukin-1 (IL-1). It stimulates a fast response by triggering phagocytosis and neutrophils mobilization, among other inflammatory mechanisms. There is a clear synergistic effect seen for a combination with the first branched (1,3)(1,6)-β-glucan and the second branched (1,3)(1,6)-β-glucan (i.e. ABB i16 which is a combination of ABB i24 and ABB i25). In order to investigate which receptors and their downstream signaling molecules could influence the strong stimulatory effect on the trained innate immunity induced by ABB i16 as follows. Monocytes were incubated in the presence of different receptor and pathway inhibitors (and appropriate negative controls) one hour prior the addition of ABB i16 (at a concentration of 1 µg / mL). The trained innate immunity phenotype was assessed by the ability of trained monocytes to release TNF-α and IL-6 after LPS re-challenge on day 6. The blockade of the uptake of ABB i16 by Dectin-1, CR3 and TLR4 led to a significant decrease in the cytokine production of both TNF-α and IL-6 after LPS restimulation, in comparison with their isotype controls (Figure 4 A). The exposure of monocytes to the Syk inhibitor during the trained immunity induction phase of the experiment led to a significant decrease in the later induction of TNF-α and IL-6 in ABB i16- trained macrophages compared to the control condition (Figure 4 B). A similar, though less pronounced downregulation of TNF-α and IL-6 production was observed after blocking the Dectin-1 signaling pathway component Raf-1 (Figure 4 B). Subsequently, it was assessed whether the inhibition of the PI3K signaling cascade also plays a role in trained innate immunity induction by ABB i16. It is known in the art that β-glucan recognition in innate immune cells by Dectin-1 and subsequent intracellular signaling can be mediated by other downstream components, such as phosphoinositide-3 kinase (PI3K) (Shah et al., 2009. Molecular Immunology 46: 8-9). Indeed, after stimulation of monocytes pre-incubated with a PI3K inhibitor, the production of TNF-α and IL-6 pro-inflammatory cytokines was notably lower (Figure 4 B). These results show that Dectin-1 receptor and its downstream signaling molecules Syk and Raf-1 are required for the production of IL-6 and TNFα induced by stimulation with ABB i16. Furthermore, the data show a role for the PI3K downstream mediator and CR3 in signaling following recognition of ABB i16, reflecting the complexity of the Dectin-1 signaling that plays a role in the induction of the trained innate immunity by ABB i16. Interestingly, induction of the trained innate immunity by ABB i16 also seems to be dependent on TLR4, which is in contrast to induction of the trained innate immunity by Ca β-glucans. Therefore, it might be that the observed high effectivity of the induction of the trained innate immunity by ABB i16, which is a combination of two different β-glucans ABB i24 and ABB i25, is due to dual induction of Dectin- 1 and TLR4. In addition to Dectin-1, CR3 and TLR4, a role for the MMR receptor was also observed (Figure 4 C). The inhibition of Dectin-1, MMR, CR3, and TLR4 all blocked the uptake of ABB i16, leading to a significant decrease in both TNF and IL-6 production in the first 24 hours after ABB i16 stimulation, in comparison to their corresponding controls (Figure 4 C). Example 3: β-glucan-mediated protection in cancer through innate immunity training Materials and methods A murine bladder cancer model was established as follows. Mice were anesthetized with 2-4% isofluorane. Subsequently, a catheter was inserted through the urethra into the bladder. The bladder was prepared for tumour implantation by instillation of 50 µl poly-lysine for 20 minutes. Hereafter, the catheter was removed and a new (second) catheter was inserted in the urethra for intravesical instillation of MB49 cells, BCG and / or ABB i16. Tumour challenge took place at day 0 with 7.5x105MB49 cells. In some experimental groups, at day 7 before tumour challenge (i.e. day -7) training of the innate immune system was induced by administering 1 mg of ABB i16 (or PBS in the control group). A similar experimental design was used in a murine melanoma model. In this model, tumour challenge took place at day 0 with 3x105B16.F10 cells. In some experimental groups, at day 7 before tumour challenge (i.e. day -7) training of the innate immune system was induced by administering 1 mg of ABB i16 (or PBS in the control group). The weight of the tumour was measured after excision at the end of the study. Results Administration of a combination with the first branched (1,3)(1,6)-β-glucan and the second branched (1,3)(1,6)-β-glucan (i.e. ABB i16 which is a combination of ABB i24 and ABB i25) was found to mediate protection in a murine bladder cancer model and melanoma cancer model as a reduction of tumour weight was observed in both of these cancer models (figure 5A and B). In addition, tumor volume in a murine melanoma cancer model was diminished during the entire observation period (Figure 6A). Example 4: Effect of β-glucans on Th1 / Th2 / Th17 balance. Materials and methods Buffy coats (Sanquin blood bank, NL32357.091.10) from healthy volunteers were obtained. The buffy coats were diluted 1:1 with calcium / magnesium-free phosphate-buffered saline (PBS, Gibco, Life Technologies). Peripheral blood mononuclear cells (PBMCs) were isolated using density-gradient centrifugation over Ficoll-Paque solution (GE Healthcare). The cells were washed twice with cold PBS and resuspended in Dutch-modified RPMI-1640 culture medium (Roswell Park Memorial Institute, Invitrogen), which was further supplemented with gentamicin (50 μg / mL; Centrafarm), GlutaMAX (2mM; Gibco, Life Technologies), and pyruvate (10 mM; Life Technologies) (from now on referred to as RPMI+). The PBMC yields and composition were assessed using a Sysmex XN-450 hematology analyser (Sysmex Corporation). For cell stimulation experiments, 5 ×106 / mL of the PBMCs (or 5 × 105cells in 100 μL / well) were added to a round-bottom 96-well cell culture plate in RPMI+ medium which was further supplemented with 10% human pooled serum. The cells were stimulated with various stimuli combinations. Stimuli and recombinant cytokines were used at the following final concentrations: 1, 10, or 100 μg / mL of β-glucan ABB i16, i.e. the branched (1,3)(1,6)-β-glucans ABB i24 and ABB i25 as used in example 1; 1 × 106microorganisms / mL of heat-killed Candida albicans (strain UC 820); 5 µg / mL of Bacillus Calmette–Guérin (BCG- Medac, Medac); 10 ng / mL of rhIL-12 and rhIL-18, 100 ng / mL of rhIL-1 and rhIL- 23, as well as RPMI+ as a negative control. The PBMCs were incubated at 37°C, 5% CO2 and left to rest for 7 days, after which supernatants were collected and stored at -20°C until further analysis. TNF-alpha (also referred to herein as TNF), IFN-γ, IL-5, IL-17, IL-10, and IL- 22 were measured in cell culture supernatants using DuoSet ELISA kits (R&D Systems), according to the manufacturer’s instructions. Gen5 software was used in conjunction with Microsoft Excel to calculate cytokine concentrations in the original samples. Data are shown as mean ± SEM, unless otherwise indicated. Individual data points in graphs represent biological replicates, not technical repeats. The number of data points is displayed in each figure, with the n for each experiment also indicated in the figure legend. Statistical analyses were performed in GraphPad Prism (version 9.4.1, GraphPad software) and R (4.2.3). Wilcoxon matched-pairs signed rank tests were used to compute p-values unless otherwise stated. P-values below 0.05 (two-tailed) were considered statistically significant. Results In order to investigate potential variability in the Th1, Th2, and Th17 cytokine responses based on the concentration of ABBi16 β-glucan used for stimulation, a dose-response analysis was conducted. Specifically, PBMCs were exposed to 1, 10, and 100 µg / mL of ABB i16 over 7 days, after which the cytokine production in cell culture supernatants was measured (Figure 7 A). Exposure of cells to 10 µg / mL of the ABB i16 induced significantly higher production of IFN-γ, IL-10, and IL-22, compared to the other concentrations of 1 and 100 µg / mL (Figure 7 B, C and D). TNF secretion was observed in a dose-dependent manner, being significantly higher at the condition of cells stimulated with 100 µg / mL compared to both 1 and 10 µg / mL (Figure 7 B). Interestingly, the production of IL-5 appeared independent of the ABBi16 concentration used for the 7-day PBMC stimulation (Figure 7 C). Furthermore, secretion of IL-17 remained consistently low across all concentrations of ABBi16 used for stimulation (Figure 7 D). The fact that ABBi16 did not induce IL-17 production, the prototype pro-inflammatory product of Th17 responses, suggests that it can be safely used as an oral supplement for patients with autoimmune conditions. For the subsequent experiments, a dose of 10 µg / mL of ABBi16 was selected, due to the significant increase in IFN-γ, IL-10, and IL-22 secretion observed in cells treated with this concentration of stimulus. Next, the effect observed of ABB i16 on Th1, Th2, and Th17 cytokine responses was compared to the effect induced by C. albicans. Therefore, PBMCs were treated with either ABB i16 or C. albicans for 7 days, after which cytokine production capacity was measured (Figure 8 A). C. albicans induced significantly higher amounts of Th1 cell-derived IFN-γ and TNF in PBMCs after 7 days (Figure 8 B). Regarding the Th2 response, IL-5 production appeared comparable between ABB i16 and C. albicans conditions, with no significant difference observed (Figure 8 C). Conversely, the IL-10 production was significantly elevated in cells stimulated with Candida compared to those stimulated with ABBi 16 (Figure 8 C). As expected (Figure 7 D) there was a notable absence of IL-17 production in PBMCs treated with ABB i16, while Candida stimulation exhibited significant induction of IL-17 (Figure 8 D). Notably, C. albicans also induced a significantly stronger induction of IL-22 compared to ABB i16 (Figure 8 D). Collectively, this data show that ABB i16 predominantly induces a Th1 response characterized by IFN-γ and TNF secretion, along with a Th2 response marked by IL-10 production, and additionally IL-22 induction. Furthermore, it was determined whether co-stimulation with BCG and ABB i16 would enhance Th1, Th2, and Th17 cellular responses in PBMCs (Figure 9 A). ABB i16 in combination with BCG led to increased production of IFN-γ, TNF, IL-10 in comparison with BCG or ABB i16 alone and led to an increase in IL-22 in comparison with BCG alone (Figures 9 B, 9 C and 9 D). The addition of ABB i16 to BCG led to no changes in IL-5 production (Figure 9 B). IL-17 production remained low for all treatment conditions, with no significant effect on the IL-17 response after the addition of β-glucan to BCG compared to the other stimulation conditions (Figure 9 D). These findings collectively indicate a synergy between ABB i16 and BCG for Th1 polarization, while ABB i16 induces additive effects on BCG-induced IL-10 and IL-22 production. Furthermore, it was determined whether co-stimulation with certain combinations of cytokines and ABB i16 would enhance Th1, Th2, and Th17 cellular responses in PBMCs. An increased IFN-γ, IL-17 and IL-22 production was observed in cells exposed to the recombinant cytokines IL-1β+IL-23 compared to ABB i16 alone (Figure 10 B and Figure 11 A). Surprisingly, the addition of ABB i16 to cells stimulated with IL-1β+IL-23 inhibited IL-17 production, while maintaining IFN-γ and IL-22 secretion unchanged (Figure 11 A and Figure 10 B). No difference in TNF production at day 7 between the ABBi16 stimulation and the ABBi16 combined with IL-1 β and IL-23 condition was observed (Figure 10 B). However, the latter showed a significant increase in TNF secretion compared to IL-1 β +IL-23 stimulation alone, indicating that the heightened cytokine production is solely due to ABB i16 stimulation. A significant increase in IFN-γ and TNF secretion was observed when IL-18+IL-12 was supplemented with ABBi16 compared to the production induced by either IL-18+IL-12 or ABBi16 alone (Figure 10 C). Of note, IL-22 production remained similar in cells stimulated with IL-18+IL-12 alone or in combination with ABBi16 (Figure 10 C).

Claims

Claims 1. A physiologically acceptable composition for use in the treatment of a cancer, wherein the physiologically acceptable composition comprises a first branched (1,3)(1,6)-β-glucan and a second branched (1,3)(1,6)-β-glucan different from the first.

2. The physiologically acceptable composition for use according to claim 1, wherein the first branched (1,3)(1,6)- β-glucan is free of 3-linked mannose units and / or free of 4,6 linked glucose units.

3. The physiologically acceptable composition for use according to claim 1 or claim 2, wherein at least one branched (1,3)(1,6)-β-glucan is composed of glucose units and one or more further monosaccharide units different from a glucose unit, which further monosaccharide unit(s) form up to 3 % of the monosaccharide units of said branched (1,3)(1,6)-β-glucan, preferably 1 – 3 % of the monosaccharide units of said branched (1,3)(1,6)-β-glucan, most preferably about 2 % of the monosaccharide units of said branched (1,3)(1,6)-β-glucan.

4. The physiologically acceptable composition for use according to claim 3, wherein said glucan composed of glucose units and one or more further monosaccharide units is a mannoglucan, wherein preferably the glucose content of the mannoglucan is 95.5 - 99.5% and the mannose content of the mannoglucan is 0.5 - 4.5 %, more preferably the glucose content is 97.5 - 98.5% and the mannose content is 1.5 - 2.5 %, wherein the glucose content is calculated as the number of glucose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan and the mannose content is calculated as the number of mannose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan.

5. The physiologically acceptable composition for use according to claim 4, wherein the first branched (1,3)(1,6)-β-glucan has a lower mannose content than the second branched (1,3)(1,6)-β-glucan, preferably the mannose content of the first glucan being in the range of 0 - 1.5% and the relative mannose content of the second glucan preferably being more than 1.5%, in particular up to 6 %, wherein the mannose content is calculated as the number of mannose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan.

6. The physiologically acceptable composition for use according to any of the previous claims, wherein the first branched (1,3)(1,6)-β-glucan has a lower 4,6 linked glucose content than the second branched (1,3)(1,6)-β-glucan, wherein preferably the first branched (1,3)(1,6)-β-glucan preferably has a 4,6 linked glucose content of 0 - 0.5 % and / or the second branched (1,3)(1,6)-β-glucan has a 4,6 linked glucose content of more than 0.5 %, in particular up to 2 %, wherein the 4,6 linked glucose content is calculated as the number of 4,6 linked glucose units over the total number of monosaccharide units in the branched (1,3)(1,6)-β-glucan.

7. The physiologically acceptable composition for use according to any of the previous claims, wherein the second branched (1,3)(1,6)-β-glucan has 2-10 %, preferably 3-8 %, in particular 3-7 % 3,6 linked glucose units of the total number of monosaccharide units in the β-glucan.

8. The physiologically acceptable composition for use according to any of the previous claims, wherein the first β-glucan has 2-20 %, preferably 6-16 %, more preferably 8-14 %, in particular 10.5-12.5 % 1,6 linked glucose units of the total number of glucose units in the first branched (1,3)(1,6)-β-glucan.

9. The physiologically acceptable composition for use according to any of the previous claims, wherein the second branched (1,3)(1,6)-β-glucan has 2-16 %, preferably 4-14 %, more preferably 6-12 %, in particular 8.5-10.5 % 1,6 linked glucose units of the total number of monosaccharide units in the second branched (1,3)(1,6)-β-glucan.

10. The physiologically acceptable composition for use according to any of the previous claims, wherein the second branched (1,3)(1,6)-β-glucan has more than 0 %, preferably 0.5-6 %, more preferably 1-5 %, in particular 2-4 % 2,3,4 linked glucose units of the total number of monosaccharide units in the second branched (1,3)(1,6)-β-glucan.

11. The physiologically acceptable composition for use according to any of the previous claims, further comprising at least one microorganism or a component (other than a branched (1,3)(1,6)-β-glucan), a lysate or an extract thereof, wherein the at least one microorganism preferably is a yeast.

12. The physiologically acceptable composition for use according to any of the previous claims, further comprising a mannan, wherein the content of the mannan preferably is 0.5-10 wt% based on the total weight of the composition, morepreferably the content of the mannan is 0.5-4.5 wt% based on the total weight of the composition.

13. The physiologically acceptable composition for use according to any of the previous claims, wherein the first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β-glucan bind to at least two receptors chosen from the group consisting of dectin-1, TLR-4, and CR3, preferably the first branched (1,3)(1,6)-β- glucan and second branched (1,3)(1,6)-β-glucan bind to dectin-1, TLR-4 and CR3.

14. The physiologically acceptable composition for use according to claim 13, wherein the first branched (1,3)(1,6)-β-glucan and second branched (1,3)(1,6)-β- glucan bind to at dectin-1, TLR-4, and CR3.

15. The physiologically acceptable composition for use according to any of the previous claims, wherein the first branched (1,3)(1,6)-β-glucan is ABB i25 and the second branched (1,3)(1,6)-β-glucan is ABB i24.

16. The physiologically acceptable composition for use according to any of the previous claims, wherein the content of the first and second branched (1,3)(1,6)-β- glucan together is between 40-90 wt% based on the total weight of the composition 17. The physiologically acceptable composition for use according to any of the previous claims, wherein the content of the first and second branched (1,3)(1,6)-β- glucan together is between 40-90 wt% based on the dry weight of the composition.

18. The physiologically acceptable composition for use according to any of the previous claims, wherein the cancer is bladder cancer or a melanoma.

19. The physiologically acceptable composition for use according to any of the previous claims , wherein the physiologically acceptable composition further comprises BCG vaccine.

20. The physiologically acceptable composition for use according to any of the previous claims, wherein the immune system is trained, in particular the innate immune system.

21. The physiologically acceptable composition for use according to any of previous claims, wherein the composition is to be administered directly to a specific site where cancer cells or precancerous cells are present.

22. The physiologically acceptable composition for use according to any one of previous claims, wherein the composition is to be administered orally, topically, intradermally, transdermally, parenterally, subcutaneously, intramuscularly, intraperitoneally, intrabursally, intranasally, intracavitary, intracapsulary,intraocularly, intraarterially, intralesionally, intratumorally, intravesically or by application to mucosa, preferably topically, orally, intravesically, subcutaneously or intratumorally.

23. The physiologically acceptable composition for use according to any of previous claims, wherein the treatment comprises a reduction in volume, weight or both of a malignant tumour.

24. The physiologically acceptable composition for use according to any of previous claims, wherein the treatment comprises an increase in the life- expectancy or prognosis of survival.

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