Patent Description:
Exposure of humans and animals to environmental factors such as food additives, drugs, industrial chemicals and bacterial and fungal metabolites, particularly early in life, can detrimentally affect health. The immune system is known to be most sensitive to chemical-induced toxicity. The mycotoxin deoxynivalenol (DON) is a highly prevalent food contaminant known to induce immunotoxicity in humans and animals. DON is produced as secondary metabolite from Fusarium fungi species, which contaminates human food at a global level, especially cereal and grain-based products. Acute and chronic exposure to DON can have a significant impact on growth and food consumption, and negatively affect intestinal, neurological and reproductive systems. The immune system is extremely sensitive to DON, since ingestion of very low levels can induce immunomodulatory effects.

In human milk, human milk oligosaccharides (HMOs) are present that provide health benefit to infants such as improvement of the intestinal microbiota, the intestinal gut barrier and the innate mucosal immune system. Non-digestible oligosaccharides such as galactooligosaccharides (GOS) comprising galactosyllactoses (GLs), such as <NUM>'-GL, <NUM>'-GL and <NUM>'-GL, which are formed by the elongation of lactose with a further galactose residue, forming different galactosyl-lactoses can have similar functional properties as (HMOs). The composition of those GLs generated by trans-glycosylation highly depends on the enzyme source and technology chosen.

<CIT> shows that GOS/GLs have an anti-inflammatory properties on human intestinal epithelial cells in vitro and an GL-specific improved gut barrier effect.

<CIT> discloses a composition comprising a non-digestible oligosaccharide for use in the treatment, prevention or alleviation of a condition that has resulted from exposure to a trichothecene mycotoxin exposure in an individual, wherein the non-digestible oligosaccharide has a degree of polymerisation (DP) of <NUM> - <NUM>.

<CIT> discloses a composition for use in reducing the risk of occurrence of or preventing whey protein allergy associated with mycotoxin exposure in an infant who consumes cereals or cereal-comprising products daily and suffers from an increased risk of whey protein allergy associated with mycotoxin exposure, said composition comprising non-digestible oligosaccharide. <CIT> is silent on any effect of DON on systemic and adaptive immune response to pathogens or a vaccine or antigen.

<NPL> discloses a study directed toward the effect of beta1,<NUM>'-galactosyllactose toward gut barrier integrity in cell cultures exposed to DON. <CIT> concerns a nutritional composition comprising a combination of beta <NUM>,<NUM> galacto-oligosaccharides and beta <NUM>,<NUM> and/or <NUM>,<NUM> galacto-oligosaccharides and superior effects on immune system are demonstrated. <CIT> is directed toward the use of a non-digestible oligosaccharide for providing nutrition to an infant suffering from an increased risk of food allergy. The infant is preferably at increased risk of mycotoxin exposure, for instance by consuming cereals. <NPL>, relates to a study assessing the impact of GOS on DON-induced epithelial dysfunction of Caco-<NUM> cells and in mice. <CIT> discloses a composition comprising non-digestible oligosaccharide including GOS for treatment, prevention or alleviation of a mycotoxin exposure associated condition.

The mycotoxin deoxynivalenol (DON) is known to disrupt intestinal barrier and induce immunotoxicity Therefore, the effects of dietary interventions with trans-galactosyl-oligosaccharides (GOS) enriched in beta3'-GL (beta1 ,<NUM>'-galactosyllactose) on DON-induced immunotoxicity in a murine Influenza vaccination model was investigated. Mice received diets containing DON, GOS or a combination of these two.

DON exposure decreased Tbet+ Th1 cells in the spleen and induced significant reduction in IFN-γ secretion from splenocytes after ex vivo re-stimulation. Addition of GOS in DON-contaminated diets increased the frequency of Tbet+ Th1 cells and secretion of IFN-γ from re-stimulated splenocytes, and therefore prevent DON-induced reduction in type-<NUM> immune responses in vaccinated animals. Moreover, consuming DON-contaminated diets caused a significant drop in frequency of B cells in the spleen of vaccinated mice, which corresponds to the reduction in vaccine-specific IgG production in these animals. Addition of GOS to the diet of DON-exposed mice restored the percentage of B cells to the values of the control group.

It thus was found that that supplementation with GOS comprising beta3'-GL could restore B cells in the spleen and improve vaccination responsiveness in DON-exposed mice.

The invention concerns galacto-oligosaccharides or a nutritional composition comprising galacto-oligosaccharides for use in therapeutically preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus, wherein the galacto-oligosaccharides comprise beta1,<NUM>'-galactosyllactose, and wherein the galacto-oligosaccharides comprise at least <NUM> % beta1,<NUM>'-galactosyllactose based on total weight of the galacto-oligosaccharides.

The nutritional composition is a synthetic nutritional composition.

According to the present invention, beta1,<NUM>'-galactosyllactose is seen as the active component for achieving the advantageous effect on prevention and/or amelioration of a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus.

Likewise the invention can be worded as beta1,<NUM>'-galactosyllactose for use in preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus.

Mycotoxins are secondary metabolites produced by moulds and fungi contaminating cereal grains as well as forages, fruits, feed and food products as well as the environment (e.g., soil, water and air through aerosol acquired mycotoxicosis, etc.). Mycotoxins may have dangerous effects on human and animal health. Of particular note are the trichothecene mycotoxins, which are a class of compounds produced by the species Fusarium graminearum. This large family of sesquiterpene epoxides are closely related and vary by the position and number of hydroxylations and substitutions of a basic chemical structure. The major trichothecene produced by Fusarium graminearum is deoxynivalenol (DON) also known as vomitoxin for its ability to induce vomiting. The impact of DON on nutrient absorption in human intestinal epithelial cells has been investigated in <NPL>, and in <NPL>.

Mycotoxins can appear in the food chain as a result of fungal infection of plant products (e.g., forage, grain, plant protein, processed grain by-products, roughage and molasses products), and can either be eaten directly by humans, or introduced by contaminated grains, livestock or other animal feedstuff(s). Since DON frequently occurs in toxicologically relevant concentrations in cereals and grains, it can be qualified as a genuine problem for all humans and animals consuming a diet comprising cereals and/or grains. It is a particular concern for infants, and with that in mind Codex Committees on Contaminants in Food (CCCF) have been dedicated to provide maximum limits for deoxynivalenol levels still deemed acceptable in raw cereal grains such as wheat and barley grain and infant formula. DON can also be present in the milk produced by lactating mothers consuming a diet contaminated with DON.

It was now found that upon consumption of diets contaminated with DON, a significant drop in frequency of B cells in the spleen of vaccinated mice was observed. The drop in frequency of B cells corresponded to the reduction in vaccine-specific IgG production. Addition of GOS that comprised beta1,<NUM>'-galactosyllactose to the diet of DON-exposed mice restored the percentage of B cells. Also DON contamination in the diet induced a reduction in Tbet+ Th1 cells in the spleen of vaccinated mice. The addition of GOS that comprised beta1,<NUM>'-galactosyllactose to the diet increased the percentage of Tbet+ Th1 cells in the spleen of DON-exposed mice and restored IFN-γ production. Hence the use of GOS that comprise beta1,<NUM>'-galactosyllactose attenuate the adverse effects of DON on systemic adaptive immune response.

The present invention relates to beta1,<NUM>'-galactosyllactose, herein also referred to as beta3'-GL, for use in preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus.

This beta1,<NUM>'-galactosyllactose can be administered as such, in a suitable matrix, or in a nutritional composition. The beta1 ,<NUM>'-galactosyllactose may for example be part of a mixture of galacto-oligosaccharides (GOS), preferably beta-galacto-oligosaccharides (betaGOS). Beta1,<NUM>'-galactosyllactose is Gal-(beta <NUM>,<NUM>)-Gal-(beta <NUM>,<NUM>)-Glc, wherein Gal stands for galactose and Glc for glucose.

In a preferred embodiment, the beta1,<NUM>'-galactosyllactose is used as such. In another preferred embodiment, the beta1,<NUM>'-galactosyllactose is present in a nutritional composition. In one embodiment, the invention relates to galacto-oligosaccharides or to a nutritional composition comprising galacto-oligosaccharides for use in preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response, wherein the galacto-oligosaccharides comprises beta1,<NUM>'-galactosyllactose, and wherein the galacto-oligosaccharides comprise at least <NUM> % beta1 ,<NUM>'-galactosyllactose based on total weight of the galacto-oligosaccharides, wherein the nutritional composition is a synthetic nutritional composition.

The nutritional composition comprising beta1,<NUM>'-galactosyllactose or the nutritional composition comprising galacto-oligosaccharides that comprise beta1,<NUM>'-galactosyllactose for use according to the invention are herein also referred to as the present nutritional composition, or nutritional composition according to the present invention, or final nutritional composition. The nutritional composition according to the present invention is not human milk.

As described above, the beta1,<NUM>'-galactosyllactose may be part of a mixture of galacto-oligosaccharides (GOS), preferably beta-galacto-oligosaccharides (betaGOS).

A suitable way to form GOS is to treat lactose with beta-galactosidases. Dependent on the specificity of the enzyme used, a galactose unit is hydrolysed from lactose and coupled to another lactose unit via a beta-linkage to form a trisaccharide. A galactose unit may also be coupled to another single galactose unit to form a disaccharide. Subsequent galactose units are coupled to form oligosaccharides. The majority of such formed oligosaccharides have a degree of polymerization (DP) of <NUM> or lower. Depending on the enzyme these linkages between the galactose residues can be predominantly beta1,<NUM>', beta1,<NUM>' or beta1,<NUM>'.

A suitable way to produce beta1,<NUM>'-galactosyllactose, is by using a beta-galactosidase from S. thermophilus. Particularly suitable is the use of beta-galactosidase from strain CNCM I-<NUM> and/or CNCM I-<NUM> in a process as disclosed in example <NUM> of <CIT> or example <NUM> of <CIT>. thermophilus CNCM I-<NUM> was deposited under the Budapest Treaty on <NUM> August <NUM> at Collection Nationale de Cultures de Microorganisms van Institute Pasteur, Paris, France by Compagnie Gervais Danone. thermophilus CNCM I-<NUM> is also referred to as strain S. thermophilus ST065. thermophilus CNCM I-<NUM> was deposited under the Budapest Treaty on <NUM> August <NUM> at Collection Nationale de Cultures de Microorganisms van Institute Pasteur, Paris, France by Compagnie Gervais Danone. Both strains have also been published in <CIT>. The composition of this GOS is also described in more detail in <NPL> and also in and in example <NUM> of <CIT>. The amount of beta1,<NUM>'-galactosyllactose in this GOS preparation is in the range of <NUM>-<NUM> wt%, based on total galacto-oligosaccharides (excluding lactose, galactose and glucose). Another preferred sources of beta1,<NUM>'-galactosyllactose are commercially available GOS rich in beta1,<NUM> and beta1,<NUM> galacto-oligosaccharides include Bimuno from Clasado, or Purimune from GTC Nutrition. Beta1,<NUM>'- and beta1,<NUM>'-galactosyllactose can be enriched or purified from these GOS mixtures as known in the art, for example by size exclusion chromatography. Alternatively, pure beta1,<NUM>'-galactosyllactose is commercially available (Carbosynth Ltd, Compton, UK).

The GOS, including betaGOS, are non-digestible. Human digestive enzymes (including human lactase) are not able to hydrolyse GOS. GOS when consumed therefore reaches the large intestine intact and is available for fermentation by the intestinal microbiota.

Preferably the nutritional composition according to the present invention comprises at least <NUM> GOS per <NUM>, more preferably at least <NUM> even more preferably at least <NUM> per <NUM>. Preferably the composition does not comprise more than <NUM> of GOS per <NUM>, preferably not more than <NUM>, more preferably not more than <NUM>. More preferably, the nutritional composition according to the present invention comprises GOS in an amount of <NUM> to <NUM>/<NUM>, even more preferably in an amount of <NUM> to <NUM>/<NUM>, even more preferably in an amount of <NUM> to <NUM>/<NUM>.

In a preferred embodiment, the nutritional composition comprising galacto-oligosaccharides for use according to the present invention comprises at least <NUM> wt% galacto-oligosaccharides based on dry weight of the nutritional composition. Preferably the nutritional composition according to the present invention comprises at least <NUM> wt. % GOS based on dry weight of the nutritional composition, more preferably at least <NUM> wt. %, even more preferably at least <NUM> wt. %, all based on dry weight of the nutritional composition. Preferably the nutritional composition does not comprise more than <NUM> wt. % of GOS based on dry weight of the nutritional composition, more preferably not more than <NUM> wt. %, even more preferably not more than <NUM> wt%. The nutritional composition according to the present invention preferably comprises GOS in an amount of <NUM> to <NUM> wt. %, more preferably in an amount of <NUM> to <NUM> wt. %, most preferably in an amount of <NUM> to <NUM> wt. %, all based on dry weight of the nutritional composition.

Preferably the nutritional composition according to the present invention comprises at least <NUM> GOS per <NUM> kcal, more preferably at least <NUM>, even more preferably at least <NUM> per <NUM> kcal. Preferably the composition does not comprise more than <NUM> of GOS per <NUM> kcal, preferably not more than <NUM> per <NUM> kcal, more preferably not more than <NUM> per <NUM> kcal. More preferably, the nutritional composition according to the present invention comprises GOS in an amount of <NUM> to <NUM> per <NUM> kcal, even more preferably in an amount of <NUM> to <NUM> per <NUM>, even more preferably in an amount of <NUM> to <NUM> per <NUM>. Lower amounts result in a less effective composition, whereas the presence of higher amounts of GOS may result in side-effects such as osmotic disturbances, abdominal pain, bloating, gas formation and/or flatulence.

The nutritional composition comprising galacto-oligosaccharides for use according to the present invention comprises at least <NUM> % beta1 ,<NUM>'-galactosyllactose based on total weight of the galacto-oligosaccharides, preferably the nutritional composition comprising galacto-oligosaccharides comprises at least <NUM> % beta1 ,<NUM>'-galactosyllactose based on total weight of the galacto-oligosaccharides.

It is advantageous to have general GOS present in the present nutritional composition, besides the specific beta1 ,<NUM>'-galactosyllactose. A mixture of GOS with different sizes and linkages will have an increased beneficial effect on the microbiota and an improved production of short chain fatty acids, which in its turn have a further improved beneficial effect on immune function.

The total amount of GOS as defined for the present nutritional composition is including the amount of beta1,<NUM>'-galactosyllactose.

In a preferred embodiment, the nutritional composition according to the present invention comprises <NUM> to <NUM> galacto-oligosaccharides per <NUM>, wherein <NUM> to <NUM> per <NUM> of the galacto-oligosaccharides is beta1,<NUM>'-galactosyllactose. In another preferred embodiment, the nutritional composition according to the present invention comprises <NUM> to <NUM> galacto-oligosaccharides per <NUM>, wherein the amount of beta1,<NUM>'-galactosyllactose is more than <NUM> wt% based on total galacto-oligosaccharides, preferably more than <NUM> wt% based on total galacto-oligosaccharides. In another preferred embodiment, the nutritional composition according to the present invention comprises <NUM> to <NUM> g galacto-oligosaccharides per <NUM>, wherein the amount of beta1,<NUM>'-galactosyllactose ranges from <NUM>-<NUM> per <NUM>. In another preferred embodiment, the nutritional composition according to the present invention comprises <NUM> to <NUM> galacto-oligosaccharides per <NUM>, wherein the amount beta1,<NUM>'-galactosyllactose is more than <NUM> wt% based on total galacto-oligosaccharides, preferably more than <NUM> wt% based on total galacto-oligosaccharides, and wherein the amount of beta1,<NUM>'-galactosyllactose is between <NUM> and <NUM> per <NUM>.

In a preferred embodiment, the nutritional composition according to the present invention comprises <NUM> to <NUM> wt%, more preferably <NUM> to <NUM> wt. %, beta1,<NUM>'-galactosyllactose, based on dry weight of the nutritional composition. In a preferred embodiment, the nutritional composition comprises <NUM> to <NUM> wt% beta1,<NUM>'-galactosyllactose, based on dry weight of the nutritional composition. In another preferred embodiment, the nutritional composition comprises <NUM> to <NUM> wt% beta1,<NUM>'-galactosyllactose based on dry weight of the nutritional composition.

The nutritional composition according to the present invention preferably comprises <NUM> to <NUM> beta1,<NUM>'-galactosyllactose per <NUM> kcal of the nutritional composition. In a preferred embodiment, the nutritional composition comprises <NUM> to <NUM> beta1,<NUM>'-galactosyllactose per <NUM> kcal of the nutritional composition. In another preferred embodiment, the nutritional composition comprises <NUM> to <NUM> beta1,<NUM>'-galactosyllactose, per <NUM> kcal of the nutritional composition.

The nutritional composition according to the present invention preferably comprises <NUM> to <NUM> beta1,<NUM>'-galactosyllactose, per <NUM> of the nutritional composition. In a preferred embodiment, the nutritional composition comprises <NUM> to <NUM>, preferably <NUM> to <NUM> beta1,<NUM>'-galactosyllactose, per <NUM> of the nutritional composition. In another preferred embodiment, the nutritional composition comprises <NUM> to <NUM> beta1 ,<NUM>'-galactosyllactose per <NUM> of the nutritional composition.

Both when the beta1,<NUM>'-galactosyllactose is used as such and when it is present in a nutritional composition, it is preferred that the beta1,<NUM>'-galactosyllactose is administered in a daily dose of at least <NUM>, preferably at least <NUM> or <NUM>. It is preferred that the maximum daily dose is <NUM>, preferably <NUM>, <NUM> or <NUM>. Preferably the beta1,<NUM>'-galactosyllactose is administered in a daily dose of <NUM> to <NUM>, more preferred <NUM> to <NUM>.

It is preferred that the nutritional composition according to the present invention comprises <NUM> to <NUM> wt%, more preferably <NUM> to <NUM> wt. %, beta1,<NUM>'-galactosyllactose, based on dry weight of the nutritional composition and that the beta1,<NUM>'-galactosyllactose is administered in a daily dose of <NUM> to <NUM>. Preferably the beta1,<NUM>'-galactosyllactose is administered in a daily dose of <NUM> to <NUM>, more preferred <NUM> to <NUM>.

Preferably the nutritional composition according to the present invention also comprises fructo-oligosaccharides (FOS), as described in more detail below.

Preferably the nutritional composition according to the present invention further comprises a source of protein, a source of lipids and a source of and digestible carbohydrates, as described in more detail below.

When the beta1,<NUM>'-galactosyllactose is present in a nutritional composition, in a preferred embodiment the nutritional composition is an infant formula, a follow on formula or a young child formula.

Preferably the beta1,<NUM>'-galactosyllactose is present in a nutritional composition. The nutritional composition according to the present invention is not human milk. The nutritional composition according to the present invention is not a natural milk, for example cow's milk. The nutritional composition is a synthetic nutritional composition.

The present nutritional composition is preferably enterally administered, more preferably orally.

The present nutritional composition is preferably an infant formula, a follow on formula or a young child formula.

Examples of a young child formula are toddler milk, toddler formula and growing up milk. More preferably the nutritional composition is a follow on formula or young child formula. An infant formula is defined as a formula for use in infants and can for example be a starter formula, intended for infants of <NUM> to <NUM> or <NUM> to <NUM> months of age. A follow on formula is intended for infants of <NUM> or <NUM> months to <NUM> months of age. At this age infants start weaning on other food. A young child formula, or toddler or growing up milk or formula is intended for children of above <NUM> months of age preferably up to <NUM> months of age. Preferably the present nutritional composition is a follow on formula or young child formula.

The present nutritional composition preferably comprises lipid, protein and carbohydrate and is preferably administered in liquid form. The present nutritional composition may also be in the form of a dry food, preferably in the form of a powder which is accompanied with instructions as to mix said dry food, preferably powder, with a suitable liquid, preferably water. The present nutritional composition may thus be in the form of a powder, suitable to reconstitute with water to provide a ready to drink nutritional composition, preferably a ready to drink infant formula, follow on formula or young child formula, more preferably a ready to drink follow on formula or young child formula. The nutritional composition according to the invention preferably comprises other fractions, such as vitamins, minerals, trace elements and other micronutrients in order to make it a complete nutritional composition. Preferably infant formulae comprise vitamins, minerals, trace elements and other micronutrients according to international directives.

The present nutritional composition preferably comprises lipid, protein and digestible carbohydrate wherein the lipid provides <NUM> to <NUM>% of the total calories, the protein provides <NUM> to <NUM>% of the total calories, and the digestible carbohydrate provides <NUM> to <NUM>% of the total calories. Preferably, in the present nutritional composition the lipid provides <NUM> to <NUM>% of the total calories, the protein provides <NUM> to <NUM>% of the total calories, and the digestible carbohydrate provides <NUM> to <NUM>% of the total calories. For calculation of the % of total calories for the protein, the total of energy provided by proteins, peptides and amino acids needs to be taken into account.

Preferably the lipid provides <NUM> to <NUM> lipid per <NUM> kcal, preferably <NUM> to <NUM> per <NUM> kcal, the protein provides <NUM> to <NUM> per <NUM> kcal, preferably <NUM> to <NUM> per <NUM> kcal and the digestible carbohydrate provides <NUM> to <NUM> per <NUM> kcal, preferably <NUM> to <NUM> per <NUM> kcal of the nutritional composition. Preferably the present nutritional composition comprises lipid providing <NUM> to <NUM> per <NUM> kcal, protein providing <NUM> to <NUM> per <NUM> kcal and digestible carbohydrate providing <NUM> to <NUM> per <NUM> kcal of the nutritional composition.

Preferably the lipid provides <NUM> to <NUM> lipid per <NUM>, preferably <NUM> to <NUM> per <NUM>, the protein provides <NUM> to <NUM> per <NUM>, preferably <NUM> to <NUM> per <NUM> and the digestible carbohydrate provides <NUM> to <NUM> per <NUM>, preferably <NUM> to <NUM> per <NUM> of the nutritional composition. Preferably the present nutritional composition comprises lipid providing <NUM> to <NUM> per <NUM>, protein providing <NUM> to <NUM> per <NUM> and digestible carbohydrate providing <NUM> to <NUM> per <NUM> of the nutritional composition.

Preferably the lipid provides <NUM> to <NUM> wt%, preferably <NUM> to <NUM> wt. %, based on dry weight of the composition, the protein provides <NUM> to <NUM> wt%, preferably <NUM> to <NUM> wt. %, based on dry weight of the composition and the digestible carbohydrates comprise <NUM> to <NUM> wt%, preferably <NUM> to <NUM> wt. %, based on dry weight of the composition. Preferably the present nutritional composition comprises lipid providing <NUM> to <NUM> wt. %, protein providing <NUM> to <NUM> wt. % and digestible carbohydrate providing <NUM> to <NUM> wt. %, all based on dry weight of the composition.

The present composition preferably comprises lipids. Preferably the present composition comprises at least one lipid selected from the group consisting of vegetable lipids. Preferably the present composition comprises a combination of vegetable lipids and at least one oil selected from the group consisting of fish oil, algae oil, fungal oil, and bacterial oil. Preferably the lipid comprises the essential fatty acids alpha-linolenic acid (ALA), linoleic acid (LA) and/or long chain polyunsaturated fatty acids (LC-PUFA). The LC-PUFA, LA and/or ALA may be provided as free fatty acids, in triglyceride form, in diglyceride form, in monoglyceride form, in phospholipid form, or as a mixture of one of more of the above. Preferably the present nutritional composition comprises at least one, preferably at least two lipid sources selected from the group consisting of rape seed oil (such as colza oil, low erucic acid rape seed oil and canola oil), high oleic sunflower oil, high oleic safflower oil, olive oil, marine oils, microbial oils, coconut oil, palm kernel oil.

The present nutritional composition preferably comprises long chain poly-unsaturated fatty acids (LC-PUFA). LC-PUFA are fatty acids or fatty acyl chains with a length of <NUM> to <NUM> carbon atoms, preferably <NUM> or <NUM> carbon atoms, comprising two or more unsaturated bonds. Preferably the composition comprises n3-LCPUFA. Preferably at least one, preferably two, LC-PUFA selected from docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA). These n3-LC-PUFA are considered to support immune function and may therefore be particularly advantageously combined with beta1,<NUM>'-galactosyllactose in order to support prevention or amelioration of a food contaminant induced decrease of the adaptive immune response. This combination has unexpected advantageous effects and preferably works synergistically. The LC-PUFA may be provided as free fatty acids, in triglyceride form, in diglyceride form, in monoglyceride form, in phospholipid form, or as a mixture of one of more of the above. Suitable sources of these LC-PUFA are e.g. fish oil and oil from Mortierella alpina.

The preferred content of LC-PUFA in the present nutritional composition does not exceed <NUM> wt. % of total fatty acids, preferably does not exceed <NUM> wt. %, even more preferably does not exceed <NUM> wt. Preferably the present composition comprises at least <NUM> wt. %, preferably at least <NUM> wt. %, more preferably at least <NUM> wt. %, even more preferably at least <NUM> wt. % LC-PUFA of total fatty acids, more preferably DHA.

The present nutritional composition preferably comprises protein. The protein used in the nutritional composition is preferably selected from the group consisting of non-human animal proteins, preferably milk proteins, vegetable proteins, such as preferably soy protein and/or rice protein, and mixtures thereof. The present nutritional composition preferably contains casein, and/or whey protein, more preferably bovine whey proteins and/or bovine casein. Thus in one embodiment the protein in the present nutritional composition comprises protein selected from the group consisting of whey protein and casein, preferably whey protein and casein, preferably the whey protein and/or casein is from cow's milk. Preferably the protein comprises less than <NUM> wt% based on total protein of free amino acids, dipeptides, tripeptides or hydrolysed protein. The present nutritional composition preferably comprises casein and whey proteins in a weight ratio casein : whey protein of <NUM> : <NUM> to <NUM> : <NUM>, more preferably <NUM> : <NUM> to <NUM> : <NUM>, even more preferably <NUM> : <NUM> to <NUM> : <NUM>.

The wt% protein based on dry weight of the present nutritional composition is calculated according to the Kjeldahl-method by measuring total nitrogen and using a conversion factor of <NUM> in case of casein, or a conversion factor of <NUM> for other proteins than casein. The term 'protein' or 'protein component' as used in the present invention refers to the sum of proteins, peptides and free amino acids.

The present nutritional composition preferably comprises digestible carbohydrate. Preferred digestible carbohydrates are lactose, glucose, sucrose, fructose, galactose, maltose, starch and maltodextrin. Lactose is the main digestible carbohydrate present in human milk. The present nutritional composition preferably comprises lactose. Preferably the present nutritional composition does not comprise high amounts of carbohydrates other than lactose. Compared to digestible carbohydrates such as maltodextrin, sucrose, glucose, maltose and other digestible carbohydrates with a high glycemic index, lactose has a lower glycemic index and is therefore preferred. The present nutritional composition preferably comprises digestible carbohydrate, wherein at least <NUM> wt%, more preferably at least <NUM> wt%, more preferably at least <NUM> wt%, more preferably at least <NUM> wt%, even more preferably at least <NUM> wt%, most preferably at least <NUM> wt% of the digestible carbohydrate is lactose. Based on dry weight the present nutritional composition preferably comprises at least <NUM> wt% lactose, preferably at least <NUM> wt%, more preferably at least <NUM> wt% lactose.

The present nutritional composition preferably comprises non-digestible oligosaccharides (NDO). The term "oligosaccharides" as used herein refers to saccharides with a degree of polymerization (DP) of <NUM> to <NUM>, preferably a DP <NUM> to <NUM>, more preferably <NUM> to <NUM>, even more preferably <NUM> to <NUM>. If oligosaccharide with a DP of <NUM> to <NUM> is included in the present nutritional composition, this results in compositions that may contain oligosaccharides with a DP of <NUM> to <NUM>, a DP of <NUM> to <NUM> and/or a DP of <NUM> to <NUM>. The term "non-digestible oligosaccharides" (NDO) as used in the present invention refers to oligosaccharides which are not digested in the intestine by the action of acids or digestive enzymes present in the human upper digestive tract, e.g. small intestine and stomach, but which are preferably fermented by the human intestinal microbiota. For example, sucrose, lactose, maltose and maltodextrins are considered digestible.

Preferably the present non-digestible oligosaccharides are soluble. The term "soluble" as used herein, when having reference to a polysaccharides, fibres or oligosaccharides, means that the substance is at least soluble according to the method described by <NPL>).

Beta1,<NUM>'-galactosyllactose is considered a non-digestible oligosaccharide, more in particular a non-digestible galacto-oligosaccharide. As described above, the beta1,<NUM>'-galactosyllactose may be present in the nutritional composition according to the invention as such, or as part of a mixture of galacto-oligosaccharides (GOS), preferably beta-galacto-oligosaccharides (BGOS). In a preferred embodiment the beta1,<NUM>'-galactosyllactose is present as part of a mixture of galacto-oligosaccharides.

Galacto-oligosaccharides, and preferred embodiments of the present nutritional composition comprising galacto-oligosaccharides, are described in more detail above.

Preferably the present nutritional composition also comprises fructo-oligosaccharides (FOS). The term "fructo-oligosaccharides" as used in the present invention refers to carbohydrates composed of over <NUM>%, preferably over <NUM> % fructose units based on monomeric subunits, in which at least <NUM>%, more preferably at least <NUM>%, even more preferably at least <NUM>%, of the fructose units are linked together via a beta-glycosidic linkage, preferably a beta-<NUM>,<NUM> glycosidic linkage. A glucose unit may be present at the reducing end of the chain of fructose units. Preferably the fructo-oligosaccharides have a DP or average DP in the range of <NUM> to <NUM>, more preferably <NUM> to <NUM>, even more preferably <NUM> to <NUM>. The term "fructo-oligosaccharides" comprises levan, hydrolysed levan, inulin, hydrolysed inulin, and synthesised fructo-oligosaccharides. Preferably the preparation comprises long chain fructo-oligosaccharides with an average DP above <NUM>. Fructo-oligosaccharide suitable for use in the composition of the invention is also readily commercially available, e.g. RaftilineHP (Orafti). Preferably the nutritional composition according to the present invention comprises at least <NUM> FOS per <NUM>, more preferably at least <NUM> even more preferably at least <NUM>. Preferably the composition does not comprise more than <NUM> FOS per <NUM>, more preferably not more than <NUM> per <NUM> and most preferably not more than <NUM> per <NUM>. The amount of FOS is preferably <NUM> to <NUM> fructo-oligosaccharides per <NUM>, preferably <NUM> to <NUM> per <NUM>, more preferably <NUM> to <NUM> per <NUM>. Preferably the nutritional composition according to the present invention comprises at least <NUM> wt. % FOS based on dry weight, more preferably at least <NUM> wt. %, even more preferably at least <NUM> wt. Preferably the composition does not comprise more than <NUM> wt. % FOS based on dry weight of the total composition, more preferably not more than <NUM> wt. The presence of FOS shows a further improved effect on the microbiota and its SCFA production. The presence of fructo-oligosaccharides with a DP or average DP in the range of <NUM> to <NUM> further improves the vaccination response. It is believed that fructo-oligosaccharides with a DP or average DP in the range of <NUM> to <NUM> acts synergistically with beta1,<NUM>'-galactosyllactose on improving vaccination response.

Preferably the present nutritional composition comprises a mixture of galacto-oligosaccharides (including the beta1 ,<NUM>'-galactosyllactose) and fructo-oligosaccharides. Preferably the mixture of galacto-oligosaccharides and fructo-oligosaccharides is present in a weight ratio of from <NUM>/<NUM> to <NUM>/<NUM>, more preferably from <NUM>/<NUM> to <NUM>/<NUM>, more preferably from <NUM>/<NUM> to <NUM>/<NUM>, more preferably from <NUM>/<NUM> to <NUM>/<NUM>, more preferably from <NUM>/<NUM> to <NUM>/<NUM>, even more preferably from <NUM>/<NUM> to <NUM>/<NUM>, even more preferably in a ratio of about <NUM>/<NUM>. This weight ratio is particularly advantageous when the galacto-oligosaccharides have a low average DP and fructo-oligosaccharides has a relatively high DP. Most preferred is a mixture of galacto-oligosaccharides with an average DP below <NUM>, preferably below <NUM>, and fructo-oligosaccharides with an average DP above <NUM>, preferably above <NUM>, even more preferably above <NUM>.

The present nutritional composition preferably comprises <NUM> to <NUM> wt% total non-digestible oligosaccharides, more preferably <NUM> to <NUM> wt%, most preferably <NUM> to <NUM> wt%, based on dry weight of the nutritional composition. Based on <NUM> the present nutritional composition preferably comprises <NUM> to <NUM> total non-digestible oligosaccharides, more preferably <NUM> to <NUM>, most preferably <NUM> to <NUM>, based on <NUM> of the nutritional composition. A lower amount of non-digestible oligosaccharides will be less effective in improving the gut barrier function, whereas a too high amount will result in side-effects of bloating and abdominal discomfort. The total amount of non-digestible oligosaccharides includes galacto-oligosaccharides, including beta1,<NUM>'-galactosyllactose, fructo-oligosaccharides and any additional non-digestible oligosaccharides that may further be present in the composition.

It is also important that the nutritional composition according to the present invention does not have an excessive caloric density, however still provides sufficient calories to feed the subject. Hence, the liquid food preferably has a caloric density between <NUM> and <NUM> kcal/ml, more preferably a caloric density of between <NUM> and <NUM> kcal/ml, even more preferably between <NUM> and <NUM> kcal/ml, and most preferably between <NUM> and <NUM> kcal/ml. The infant formula, follow on formula or young child formula according to the invention is for use in providing nutrition to an infant or young child, preferably an infant.

It was found that galacto-oligosaccharides that comprise beta1 ,<NUM>'-galactosyllactose unexpectedly were capable to attenuate the adverse effects of deoxynivalenol (DON) on systemic adaptive immune response. Hence galacto-oligosaccharides that comprise beta1,<NUM>'-galactosyllactose or a nutritional composition comprising galacto-oligosaccharides that comprise beta1,<NUM>'-galactosyllactose can be used in preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus.

The present invention thus relates to galacto-oligosaccharides or a nutritional composition comprising galacto-oligosaccharides for use in preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus, wherein the galacto-oligosaccharides comprise beta <NUM>,<NUM>'-galactosyllactose, and wherein the galacto-oligosaccharides comprise at least <NUM> % beta1,<NUM>'-galactosyllactose based on total weight of the galacto-oligosaccharides, wherein the nutritional composition is a synthetic nutritional composition.

More in particular, it was found that galacto-oligosaccharides that comprise beta1,<NUM>'-galactosyllactose unexpectedly were capable to prevent the reduction in B cells due to the presence of the contaminant DON. Hence the adaptive immune response is immune response that is mediated via B cells.

Also more in particular it was found that galacto-oligosaccharides that comprise beta1,<NUM>'-galactosyllactose unexpectedly were capable to prevent the reduction of Tbet+ Th1 cells in the spleen due to the presence of the contaminant DON and that galacto-oligosaccharides that comprise beta1,<NUM>'-galactosyllactose unexpectedly were capable to prevent DON-induced reduction in IFN-γ production. Hence the B cell adaptive immune response is an immune response to an antigen that is derived from a virus.

In a preferred embodiment according to the invention, the decrease of the adaptive immune response is induced by a mycotoxin. In a preferred embodiment the mycotoxin is deoxynivalenol. In a preferred embodiment according to the invention, the decrease of the B cell adaptive immune response is induced by deoxynivalenol.

In a preferred embodiment according to the invention, the galacto-oligosaccharides or nutritional composition comprising galacto-oligosaccharides is administered to an infant or young child, preferably a weaning infant or young child. In the context of the present invention, an infant is defined as a human having an age of <NUM> to <NUM> months and a young child is defined as a human having an age of <NUM> to <NUM> months. Weaning usually starts when an infant is about <NUM> to <NUM> months. The immune system of such infants and young children are especially vulnerable to food contaminants, as the adaptive immune system is still developing and levels of food contaminant have a larger impact because infants and young children have lower body weights and the daily doe based on body weight will be higher compared to adults. Therefore especially infants and young children will benefit from the invention.

In a preferred embodiment according to the invention, the galacto-oligosaccharides or nutritional composition comprising galacto-oligosaccharides is administered or consumed together or shortly before or after consumption of cereals or cereal-comprising products. Shortly before or after means preferably refers to a period of <NUM> hours, so the present nutritional composition is preferably consumed <NUM> hours or shorter before consumption of cereals or <NUM> hours or shorter after consumption of cereals or cereal-comprising products. Preferably the present nutritional composition is consumed <NUM> hours or shorter before consumption of cereals or cereal-comprising products or <NUM> hours or shorter after consumption of cereals or cereal-comprising products. Preferably the present nutritional composition is consumed <NUM> hour or shorter before consumption of cereals or cereal-comprising products or <NUM> hours or shorter after consumption of cereals or cereal-comprising products.

In a preferred embodiment, the nutritional composition for the use according to the invention is an infant formula, follow on formula or young child formula, preferably an follow on formula or young child formula. A young child formula is nutrition intended for children having an age of above <NUM> to <NUM> months. A follow on formula is nutrition intended for infants when weaning starts to <NUM> months. In a preferred embodiment, the methods or uses according to the present invention are for healthy infants.

In a further preferred embodiment, the galacto-oligosaccharides or nutritional composition comprising galacto-oligosaccharides for use according to the present invention are for subjects that are exposed to food contaminants, more preferably for infants or young children that are exposed to food contaminants. In yet a further preferred embodiment, the galacto-oligosaccharides or nutritional composition according to the present invention are for subjects that consume cereals or cereal-comprising products. In a further preferred embodiment, the galacto-oligosaccharides or nutritional composition according to the present invention are for infants that consume cereals or for young children that consume cereals or cereal-comprising products.

In the context of the present invention the term "prevention" means "reducing the risk of" or "reducing the severity of".

In this document and in its claims, the verb "to comprise" and its conjugations is used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article "a" or "an" does not exclude the possibility that more than one of the element is present, unless the context clearly requires that there be one and only one of the elements. The indefinite article "a" or "an" thus usually means "at least one".

GOS comprising beta3'-GL was prepared using S. thermophilus ST065 (CNCM I-<NUM>) beta galactosidase after which oligosaccharides were obtained by freeze drying. The dry powder contained beta3'-GL (<NUM>/<NUM>), lactose (<NUM>/<NUM>), glucose (<NUM>/<NUM>), galactose (<NUM>/<NUM>) and other oligosaccharides (<NUM>/<NUM>). This included mainly di-tri- and tetra-saccharides. More details on the GOS production and composition are given in example <NUM> of <CIT>.

Six-week-old female C57BI/6JOlaHsd mice were purchased from Envigo (Horst, The Netherlands). Upon arrival, mice were conventionally housed under specific pathogen free (SPF) conditions with a light/dark cycle of <NUM>/<NUM> (lights on from <NUM> am-<NUM> pm) at controlled relative humidity (relative humidity of <NUM>-<NUM>%) and temperature (<NUM> ± <NUM>) with access to food and tap water ad libitum. The animals were randomly grouped as <NUM> mice per cage in filter-topped makrolon cages (<NUM>×<NUM>×<NUM>, floor area <NUM> cm2, Tecnilab- BMI , Someren, the Netherlands) with wood-chip bedding (Tecnilab- BMI , Someren in the Netherlands), and tissues (VWR, the Netherlands) were available as cage enrichment at the animal facility of Utrecht University. The animals received standard diets (pelleted food, AIN-<NUM>, Ssniff Spezialdiäten, Soest, Germany) and routine care for a week upon arrival in the animal facility, before the start of the experiments. This study was conducted in accordance with institutional guidelines for the care and use of laboratory animals established by the Animal Ethics Committee of the Utrecht University, and all animal procedures related to the purpose of the research were approved under license of the national competent authority, securing full compliance the European Directive <NUM>/<NUM>/EU for the use of animals for scientific purposes.

Semi-purified AIN-<NUM> soy protein-based diets were composed and mixed by different concentrations of DON (FERMENTEK Ltd, Jerusalem, Israel) and/or GOS comprising beta3'-GL of example <NUM> by Ssniff Spezialdiäten GmbH (Soest, Germany).

After a week of acclimatization, Animals were randomly divided into <NUM> groups (n=<NUM> per group) and received either control or modified diets. <NUM> dietary groups were formed:.

Vaccination was conducted <NUM> weeks after starting the diets, using Influvac (Abbott Biologicals B. , Weesp, The Netherlands) from season <NUM>/<NUM> as previously described (<NPL>). The mice received the primary and booster vaccinations by subcutaneous injections of <NUM>µl undiluted Influvac (containing hemagglutinin (HA) and neuraminidase antigens of three strains of myxovirus influenza, in a dose equivalent to <NUM>µg/mL HA per strain, in total <NUM>µg/ml HA). The booster vaccination was given <NUM> days after the primary vaccination. Sham group (n=<NUM>, negative control) which received injections of <NUM>µl PBS instead of vaccine was used to demonstrate the specificity of vaccine-induced response.

The animals were weighed before starting the diets (day -<NUM>) and before booster vaccination (day <NUM>). The weight gain was calculated using the formula: <MAT>.

Blood was collected at the end of the experiment by orbital extraction under anesthesia and then animals were killed by cervical dislocation. Blood samples were centrifuged (<NUM>,<NUM> rpm for <NUM>) to collect the serum and were stored at -<NUM> until analysis. To determine serum concentration of vaccine-specific antibodies, enzyme-linked immunosorbent assay (ELISA) was performed. Serum samples were incubated in <NUM>-well plates (Costar EIA/RIA plate, Alphen a/d Rijn, The Netherlands) pre-coated with <NUM>:<NUM> diluted Influvac in PBS. Final dilutions of <NUM>:<NUM> and <NUM>:<NUM> of serum samples were used for IgG1 and IgG2a measurements, respectively, and a dilution series of pooled serum that contained vaccine specific antibodies was added for standard curve calculation. For blocking nonspecific binding, the plate was incubated for <NUM> with <NUM>% BSA (Sigma, Zwijndrecht, The Netherlands) in PBS at room temperature. Anti-lgG1-biotin and anti-IgG2a-biotin (Becton Dickinson, Heerhugowaard, The Netherlands) antibodies were diluted <NUM>:<NUM> in dilution buffer (PBS with <NUM>% BSA and <NUM>% Tween). The plates were subsequently incubated with a <NUM>:<NUM> dilution of streptavidin-HRP (Biosource, Etten-Leur, The Netherlands) and optical density was measured with a Benchmark microplate reader (BioRad, Hercules, CA, USA) at a wavelength of <NUM>. Concentrations in test sera were calculated in arbitrary units (AU), relative to the standard curve.

Fresh splenocytes were isolated from spleens by crushing the tissue through <NUM> cell strainers on ice. After removing red blood cells by incubating in lysis buffer (<NUM> NH4Cl, <NUM> KHC3O, and <NUM> EDTA dissolved in <NUM> demi water and filter sterilized), splenocytes were counted and resuspended in RPMI <NUM> medium containing <NUM>% fetal bovine serum and penicillin (<NUM> U/mL)/streptomycin (<NUM>µg/mL) to reach the concentration of <NUM><NUM> cell/ml. Cells were washed in PBS/<NUM>% BSA and incubated with anti-mouse CD16/CD32 (<NUM>:<NUM> dilution in PBS/<NUM>% BSA; Mouse BD Fc Block, BD Pharmingen, San Jose, CA, USA) to block non-specific binding sites. For surface staining, cells were incubated at room temperature for <NUM> with CD4- Brilliant Violet <NUM>, CCR6-PE (BioLegend, San Diego, CA, United States), CD69-PE-Cy7, CXCR3-PE, CD25-PerCP-Cy5. <NUM>, (eBiosciences, Thermo Fisher Scientific, San Diego, CA, USA), T1ST2-FITC (MD Biosciences, St. Paul, MN, USA). Viable cells were distinguished by means of a fixable viability dye eFluor® <NUM> (eBioscience). For detecting intracellular transcription factors, cells were first fixed and permeabilized with Foxp3 Staining Buffer Set (eBioscience) according to manufacturer's protocol and then stained with Foxp3-FITC (eBioscience) and RoryT-Alexafluor <NUM> (BD Pharmingen, San Jose, CA, USA) antibodies. Results were collected with BD FACSCanto II flow cytometer (Becton Dickinson, Franklin Lakes, NJ, USA) and analyzed with FlowLogic software (Inivai Technologies, Mentone, VIC, Australia).

Bone marrow cells were isolated from femurs and tibias of healthy <NUM>-week-old C57BL/6JOlaHsd mice. Collected cells were cultured in RPMI <NUM> medium (Gibco) supplemented with <NUM>% FBS and <NUM> U/mL penicillin/streptomycin, <NUM> HEPES, <NUM> sodium pyruvate, and Eagles minimum essential medium (MEM) non-essential amino acids (all from Gibco Life Technologies) in the presence of <NUM> ng/mL GM-CSF (Prosepec, The Netherlands) for <NUM> days to obtain immature BMDC (iDC). Induced iDCs were then loaded with Influvac vaccine at a concentration of <NUM>µg/mL and incubated for <NUM> at <NUM>, <NUM>% CO2 to obtain matured DCs. iDCs treated with medium were used as negative control. Splenocytes collected from vaccinated mice were cocultured with matured DCs at <NUM>:<NUM> ratio, in <NUM>-well U-bottom culture plates for <NUM> days at <NUM>, <NUM>% CO2, with supplemented RPMI <NUM> medium (Gibco).

Cell culture supernatants were collected at day <NUM> and stored at -<NUM> until use and analyzed for the concentration of interleukin (IL)-<NUM>, IL-<NUM>, IL-<NUM>, IL-<NUM>, tumor necrosis factor (TNF)-α, macrophage inflammatory protein (MIP)-<NUM> and interferon (IFN)-γ using ProcartaPlex multiplex protein assay kit (Invitrogen, Thermo Fisher Scientific, Waltham, MA, USA) according to manufacturer's instructions.

All data were analyzed by GraphPad Prism <NUM> software (GraphPad Software, San Diego, CA, USA) using one-way ANOVA, followed by a Bonferroni's multiple comparison post hoc test for selected comparisons. Data are presented as mean ± SEM. *p < <NUM>, **p < <NUM> and ***p < <NUM> were considered statistically significant.

There was no significant difference in average weight gain between different dietary groups. There was an antigen specific response to Influvac after i. injection, as determined by vaccine-specific immunoglobulin levels in serum.

The frequency and activation status of regulatory T cells (Treg) and T helper cells (Th1 and Th2) in isolated spleen samples were studied using flow cytometry. The percentage of CD25+ FoxP3+ Treg cells was not significantly affected by addition of DON in the diet of vaccinated mice, but addition of GOS to DON-contaminated diets significantly increased the percentage of Treg cells in the spleen of these animals as compared to DON-exposed groups.

No significant effect was observed on CXCR3+ Th1 cells or T1ST2+ Th2 cells due to the presence of GOS or DON in the diet.

DON contamination induced a reduction in Tbet+ Th1 cells in the spleen of vaccinated mice, compared to control (p < <NUM>), but surprisingly addition of GOS increased the percentage of Tbet+ Th1 cells in the spleen of DON-exposed mice (p < <NUM>), and restored it back to the values of the control group, see Table <NUM>.

The frequency and activation status of B-cells in isolated spleen samples were studied using flow cytometry. Surface marker expression analysis of CD19 and CD220 revealed that DON induced significant the reduction in CD19+ B220+ B cell population (p < <NUM>). Addition of GOS to DON-contaminated diets significantly (p< <NUM>) increased the percentage of B cells in the spleen and was able to restore the effect of DON (See Table <NUM>). CD27 expression was used to distinguish between memory and naive B cells.

In order to study cytokine production capacity of immune competent cells in vaccinated mice, collected splenocytes were re-stimulated ex vivo by co-culturing the cells with antigen-loaded dendritic cells. No significant effect for dietary GOS or DON was observed on concentrations of IL-<NUM>, IL-<NUM>, IL-<NUM>, TNF-α and MIP-<NUM> in cell supernatants Dietary supplementation with GOS was able to prevent DON-induced reduction in IFN-γ production (See Table <NUM>).

Adding DON in the diet of vaccinated mice had a specifically detrimental effect on B cell-mediated humoral immunity, as indicated by the reduced vaccine-specific immunoglobulin production. DON exposure also decreased Tbet+ Th1 cells in the spleen and induced significant reduction in IFN-γ secretion from splenocytes after ex vivo re-stimulation. Results of our study showed that addition of GOS in DON-contaminated diets increased the frequency of Tbet+ Th1 cells and secretion of IFN-γ from re-stimulated splenocytes, and therefore prevent DON-induced reduction in type-<NUM> immune responses in vaccinated animals. Moreover, consuming DON-contaminated diets caused a significant drop in frequency of B cells in the spleen of vaccinated mice.

In conclusion, exposure to DON leads to modulated immune responses to vaccination. Dietary intervention with oligosaccharide mixture GOS comprising beta3'-GL attenuates the adverse effects of DON on the adaptive immune response.

A follow on formula, provided as a powder in a pack with instructions to reconstitute with water to a ready to drink milk. When reconstituted the formula contains per <NUM>:.

Claim 1:
Galacto-oligosaccharides or nutritional composition comprising galacto-oligosaccharides for use in therapeutically preventing and/or ameliorating a food contaminant induced decrease of the B cell adaptive immune response to vaccination with an antigen that is derived from a virus, wherein the galacto-oligosaccharides comprise beta1,<NUM>'-galactosyllactose, and wherein the galacto-oligosaccharides comprise at least <NUM> % beta1,<NUM>'-galactosyllactose based on total weight of the galacto-oligosaccharides, wherein the nutritional composition is a synthetic nutritional composition.