RU2794127C2 - Indigestible oligosaccharide mixture for stimulation of immune system - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: group of inventions relates to the field of food industry, namely to a nutritious composition, which is not human milk and contains fucosil-lactose and beta-galacto-oligosaccharides, which have structure of [galactose]_n-glucose, where n is equal to an integer from 2 to 60, and have more than 80% of beta-1,4 and beta-1,6 bonds; and to the use of such a composition for the provision of nutrition to children, seniors and patients with cancer, and/or patients infected with HIV, as well as for the treatment and/or prevention of development of viral infections, immune diseases, and/or a disease, which can be prevented and/or treated by enhancing Th1 response and/or Th1/Th2 balance, and for increase in response to vaccination.

EFFECT: group of inventions provides for synergetic effect on enhancing Th1 response.

10 cl, 1 tbl, 3 ex

Description

The field of technology to which the invention belongs

The present invention relates to nutrition with a mixture of indigestible oligosaccharides and, in particular, to their use for stimulating the immune system.

State of the art

Human milk fed babies have a lower incidence of infections, including viral infections, than formula fed babies. Many components of human milk are believed to include immunoglobulins (such as IgA), interleukin (IL)-1, IL-6, IL-8, IL-10, interferon-γ (IFN-γ), immunocompetent cells, transforming growth factor -β (TGF-β), lactoferrin, nucleotides and non-digestible oligosaccharides (NDO) are involved in defense against infection caused by intestinal or respiratory pathogens.

NDOs are the main constituent of human milk and are the main element of the innate immune system of human milk. Human NDOs promote the growth of a beneficial microbiota dominated by bifidobacteria and lactobacilli. It is also known that some human NDOs are able to prevent direct adhesion of pathogens and toxins.

Human milk is the preferred food for children. However, it is not always possible or desirable to breastfeed a child. In such cases, infant formulas or advanced formulas are a good alternative. These mixtures should be optimally formulated so that the positive properties of breast milk can be more closely mimicked.

WO 2007/067053 discloses an infant formula comprising the plant-derived prebiotics inulin and galacturonic acid oligosaccharide, as well as a lactose-derived transgalactooligosaccharide prebiotic to reduce infections.

WO 2005/039597 relates to the use of an acid oligosaccharide or a neutral oligosaccharide for enhancing the immune system and treating and/or preventing the development of diseases associated with the immune system.

WO 01/642255 relates to a nutritional composition containing a prebiotic to enhance the immune response.

US 6,576,251 discloses a carbohydrate mixture for enteral or parenteral dietary products consisting of (a) monosaccharide(s), (b) oligosaccharide(s) (in most hexasaccharides) and (c) polysaccharide(s) (at least heptasaccharides), where the ratio in the mixture of components a, b, c, relative to the mass, is: alpha=1, b=40-1000 and c=1-50, and containing at least 1 percent by weight of fucose, present both freely and/or in association with the oligosaccharide and/or polysaccharide. The carbohydrate blend is believed to have both nutritional and biological effects that are greater than the respective effects of the individual components.

WO 99/11773 relates to methods for producing non-human transgenic mammals that produce various oligosaccharides and glycoconjugates in their milk. The subject matter claimed in the claims of this document refers to the mammals themselves, the milk they produce, compositions containing milk, milk fractions and purified oligosaccharides, as well as glycoconjugates present in milk.

WO 2005/055944 discloses pharmacological compositions containing molecules that include a fucose group in α-1,2 bonds, α-1,3 bonds or α-1,4 bonds with a galactose group, and a pharmacologically acceptable carrier.

WO 2007/105945 relates to food or supplements for pregnant women, including water-soluble indigestible saccharides. The composition is used to improve the flora and/or immune system of a pregnant woman, to improve the immune system of a child, and to improve the intestinal flora of a child after birth.

EP 1629850 provides a method and composition for the treatment and/or prevention of respiratory infections, wherein said method comprises a galactose-containing indigestible oligosaccharide and at least 5 percent by weight of an indigestible galactose saccharide.

A lot of effort is devoted to finding additional solutions to balance and stimulate the immune system.

The essence of the invention

Human milk differs from milk derived from domestic animals in that it contains more NDOs and in that the NDOs themselves are structurally different. Human NDO is very complex because it is a heterogeneous group of over 130 different compounds with varied sugar structures. Due to their complex and polymorphic structure, large-scale synthesis is difficult. For this reason, the preparation of baby food with an NDO composition identical to human milk is still technically and economically impractical.

More recently, new technologies for the chemical synthesis of specific types of NDOs identical to human NDOs have become available, thus offering the opportunity to test the immunomodulatory capacity of specific human NDOs in in vitro and in vivo assays.

The authors have unexpectedly found that fucosyllactose (FL), an oligosaccharide present in human breast milk and having a relatively simple structure, when combined with betagalactooligosaccharides, and preferably with fructooligosaccharides and/or uronic acid oligosaccharides, gives a synergistic effect in terms of stimulating the immune system. An enhanced delayed-type hypersensitivity response characteristic of an enhanced Th1 response was observed after influenza vaccine vaccination in animals treated with the present combination of FL and betagalactooligosaccharides compared to animals treated with such components alone.

It has been found that the present combination, ie. a combination including fucosyllactose and betagalactooligosaccharides, and preferably additionally including fructooligosaccharides and/or uronic acid oligosaccharides, increases the Th1 response. The observed effect was unexpectedly higher than the sum of the effects obtained from the individual components.

The present combination is particularly effective in people with a reduced Th1 response compared to healthy adults, especially newborns, elderly people suffering from "aging" of the immune system, people with AIDS or infected with the human immunodeficiency virus, cancer patients who are undergoing courses chemotherapy, radiation therapy, and patients with cancer cachexia.

The present combination is suitable for treating and/or preventing the development of infections and/or maintaining a response to vaccination before, during and/or after vaccination.

The present combination is particularly suitable for the treatment and/or prevention of diseases that can be prevented and/or cured by enhancing the Th1 response and/or Th1/Th2 balance, especially allergies, atopic dermatitis, asthma, food allergy, allergic rhinitis (eg hay fever), dust mite allergy and other forms of hypersensitivity such as anaphylaxis and acute urticaria.

Detailed description

The present invention thus relates to a nutritional composition that is not human milk and contains fucosyllactose and betagalactooligosaccharides.

In the context of the present invention, a combination comprising fucosyllactose and betagalactooligosaccharides, and preferably additionally comprising fructooligosaccharides and/or uronic acid oligosaccharides, is also referred to as the present combination. In the context of the present invention, the term "composition according to the invention" or "this composition" or "present composition" means a composition containing the present combination.

Fucosyllactose

The present combination includes fucosyllactose. Fucosyllactose (FL) is a non-digestible oligosaccharide present in human milk. It is not present in cow's milk. It is made up of three units of monose, fucose, galactose and glucose linked together. Lactose is a galactose unit linked to a glucose unit via a beta-1,4 bond. The fucose unit is linked to the galactose unit of the lactose molecule via an alpha-1,2 bond (2'-fucosyllactose, 2'-FL) or via an alpha-1,3 bond to the glucose unit of lactose (3-fucosyllactose, 3-FL). The present composition preferably includes 2'-FL.

2'-FL, preferably α-L-Fuc-(1→2)-β-D-Gal-(1→4)-D-Glc, and 3-FL, preferably α-L-Fuc-(1→3 )-[β-D-Gal-(1→4)]-D-Glc, commercially available, for example, from Sigma-Aldrich. Alternatively, they can be isolated from human milk, for example, as described in Andersson & Donald, 1981, J Chromatogr. 211:170-1744, or may be produced by genetically modified microorganisms, for example, as described in Albermann et al., 2001, Carbohydrate Res. 334:97-103.

Preferably, the composition according to the invention contains 1 mg to 3 mg fucosyllactose per 100 ml, more preferably 10 mg to 2 g, even more preferably 20 mg to 100 mg FL per 100 ml. Based on dry weight, the present composition preferably contains from 0.007% of the mass. up to 20% wt. fucosyllactose, more preferably from 0.07% of the mass. up to 10% wt., even more preferably from 0.15% wt. up to 1% wt. A lower fucosyllactose content will be less effective in stimulating the immune system, while too high a content will result in unreasonably high prices for the product.

Betagalactooligosaccharides

The present combination includes betagalactooligosaccharides. It has been found that the presence of betagalactooligosaccharides other than FL together with FL has a synergistic effect on immune stimulation, in particular the response to vaccination.

Betagalactooligosaccharide may also be referred to as transgalactooligosaccharide. In particular, a preferred embodiment of the present composition comprises betagalactooligosaccharides ([galactose] _n -glucose; where n is an integer from 2 to 60, i.e. 2, 3, 4, 5, 6, ..., 59, 60; preferably n is selected from 2, 3, 4, 5, 6, 7, 8, 9, or 10, where the galactose units are mostly linked to each other via a beta bond. Domo Ingredients, The Netherlands) Another suitable source is Bi2Munno (Classado) Preferably, the betagalactooligosaccharides include beta-1,4 and beta-1,6 bonds Preferably, betagalactooligosaccharides have more than 80% beta-1,4 and beta-1, 6 bonds, more preferably more than 90% based on whole bonds linking monomeric carbohydrate units.

Preferably, the present composition contains additional indigestible oligosaccharides with a DP of 2 to 250, more preferably 2 to 60. The indigestible oligosaccharides are preferably at least one, and more preferably at least two, selected from the group consisting of fructooligosaccharides and uronic acid oligosaccharides. .

The group of fructooligosaccharides includes inulins. Fructooligosaccharide is an NDO comprising a chain of beta-linked fructose units with a DP or average DP of 2 to 250, more preferably 2 to 100, even more preferably 10 to 60. Fructooligosaccharide includes inulin, levan and/or a mixed type of polyfructan . A particularly preferred fructooligosaccharide is inulin. Fructooligosaccharide suitable for use in formulations is also commercially available, for example from Raftiline®HP (Orafti). Preferably, the average DP of the fructooligosaccharide is greater than 20.

The group of uronic acid oligosaccharides includes oligosaccharides of mannuronic acid, guluronic acid, galacturonic acid, alginate degradation products and pectin degradation products. Uronic acid oligosaccharides are preferably formed from degradation products of pectin. Therefore, the present composition preferably contains a pectin degradation product with a DP of 2 to 100. Preferably, the pectin degradation product is obtained from apple pectin, beet pectin and/or citrus pectin. Preferably, the uronic acid oligosaccharide is a galacturonic acid oligosaccharide.

In a preferred embodiment, the composition comprises a mixture of betagalactooligosaccharides and fructooligosaccharides selected from the group consisting of short chain fructooligosaccharides and inulin, more preferably inulin. A mixture of at least two different non-digestible oligosaccharides advantageously stimulates beneficial bacteria in the gut microbiota to further grow. Preferably, the ratio by weight of the mixture of betagalactooligosaccharides and fructooligosaccharides is in the range of 25 to 0.05, more preferably 20 to 1. Preferably, the present composition contains butagalactooligosaccharides with a degree of polymerization (DP) of 2 to 10 and/or fructooligosaccharides with a DP of 2 up to 60.

In addition to FL, most preferably the composition contains betagalactooligosaccharide, fructooligosaccharide and uronic acid oligosaccharide. Such a combination has been found to act synergistically with fucosyllactose, in particular 2'-fucosyllactose. The weight ratio of betagalactooligosaccharide:fructooligosaccharide:uronic acid oligosaccharide is preferably (20-2):1:(1-20), more preferably (20-2):1:(1-10), even more preferably (20-2) :1:(1-3), even more preferably (12-7):1:(1-2). Most preferably, the ratio by weight is about 9:1:1.1. Preferably, the weight ratio of FL to betagalactooligosaccharide, preferably TOS, is 5 to 0.05, more preferably 5 to 0.1, more preferably 2 to 0.1. Preferably, the weight ratio of FL to fructooligosaccharide, preferably inulin, is 10 to 0.05, more preferably 10 to 0.1, more preferably 2 to 0.5. Preferably, the weight ratio of FL to uronic acid oligosaccharide, preferably derived from pectin, is 10 to 0.05, more preferably 10 to 0.1, more preferably 2 to 0.5.

Preferably, the composition contains from 80 mg to 4 g of indigestible oligosaccharides, including fucosyllactose and betagalactooligosaccharides per 100 ml, more preferably from 150 mg to 2 g, even more preferably from 300 mg to 1 g of indigestible oligosaccharides per 100 ml. Based on dry weight, the composition preferably contains from 0.25% of the mass. up to 25% wt. indigestible oligosaccharides, more preferably from 0.5% of the mass. up to 10% wt., even more preferably from 1.5% wt. up to 7.5% wt. A lower amount of indigestible oligosaccharides will be less effective at stimulating beneficial bacteria in the microbiota, while a higher amount will result in the side effects of flatulence and stomach discomfort.

Nutritional composition

The present combination of FL and betagalactooligosaccharides is preferably a nutritional composition. The composition of the present invention is not human milk. The present composition is preferably administered enterally, more preferably orally.

The present composition is preferably a nutritional formula, preferably an infant formula. The present composition can advantageously be used as a nutritional supplement for children. The present composition preferably contains a lipid component, a protein component and a carbohydrate component and is preferably administered in liquid form. The present invention includes dry food, preferably powders, which are accompanied by instructions on how to mix the above dry food mixture with a suitable liquid, preferably water.

The present invention advantageously provides a composition wherein the lipid component provides 5-50% of the total calories, the protein component provides 5-50% of the total calories, and the digestible carbohydrate component provides 15-85% of the total calories. The present invention advantageously provides a composition wherein the lipid component provides 20-50% of the total calories, the protein component provides 5-30% of the total calories, and the digestible carbohydrate component provides 30-70% of the total calories. Preferably, in the present composition, the lipid component provides 35-50% of the total calories, the protein component provides 7.5-12.5% of the total calories, and the digestible carbohydrate component provides 40-55% of the total calories. To calculate these percentages of total calories for the protein component, the total energy provided by proteins, peptides, and amino acids must be taken into account.

The present composition preferably contains at least one lipid selected from the group consisting of animal lipid, excluding human lipids and vegetable lipids. Preferably, the present composition contains a combination of vegetable lipids and at least one fat selected from the group consisting of fish oil, animal oil, algal oil, fungal oil and bacterial oil. The present composition preferably contains long chain polyunsaturated fatty acids (LC-PUFA). LC-PUFAs are fatty acids or fatty acyl chains of 20-24 carbon atoms, preferably 20-22 carbon atoms, containing 2 or more unsaturated bonds. More preferably, the present composition contains eicosapetanoic acid (EPA, n=3), docosahexanoic acid (DHA, n=3) and/or arachidonic acid (ARA, n=6).

Preferably, the present composition contains at least 0.1 wt%, preferably at least 0.25 wt%, more preferably at least 0.6 wt%, even more preferably at least 0.75 wt% . LC-PUFA with 20 and 22 carbon atoms, based on total fat content.

The content of LC-PUFA, in particular LC-PUFA with 20 and 22 carbon atoms, preferably does not exceed 6 wt. -%, more preferably does not exceed 3 wt.%. from the total fat content, since it is desirable to bring the composition as close as possible to human milk. The LC-PUFA can be provided as free fatty acids, triglycerides, diglycerides, monoglycerides, phospholipids, or a mixture of one or more of the above. The present composition preferably contains from 5 to 75% of the mass. polyunsaturated fatty acids, based on total fat, preferably from 10 to 50% of the mass.

The protein used in the formula is preferably selected from the group consisting of non-human animal proteins (preferably milk proteins), vegetable proteins (preferably soy protein and/or rice protein), hydrolysates of the above substances, free amino acids and a mixture of the above substances. . The present composition preferably contains casein, whey, hydrolyzed casein and/or hydrolyzed whey protein. Preferably, the protein comprises intact proteins, more preferably intact bovine whey proteins and/or intact bovine casein proteins.

The present composition preferably contains digestible carbohydrates selected from the group consisting of sucrose, lactose, glucose, fructose, dry corn syrup, starch and maltodextrins, more preferably lactose.

In light of the foregoing, it is also important that the liquid food does not have an excess caloric density, but still provides enough calories to feed the subject. Therefore, the liquid food preferably has a caloric density of 0.1 to 2.5 kcal/ml, even more preferably a caloric density of 0.5 to 1.5 kcal/ml, most preferably 0.6 to 0.8 kcal / ml.

Preferably, the present composition contains nucleotides and/or nucleosides, more preferably nucleotides. Preferably the composition contains cytidine 5'-monophosphate, uridine 5'-monophosphate, adenosine 5'-monophosphate, guanosine 5'-monophosphate and inosine 5'-monophosphate. Preferably, the composition contains from 5 to 100, more preferably from 5 to 50 mg, most preferably from 10 to 50 mg of nucleotides and/or nucleosides per 100 g dry weight of the composition. The presence of nucleotides and/or nucleotides preferentially stimulates NK cell activity. The nucleotides and/or nucleosides are believed to act synergistically with the fucosyllactose of the present composition.

Application

The present combination of FL and betagalactooligosaccharides has been found to synergistically stimulate the immune system. In particular, the Th1 response is enhanced. The effect of combining these two components is greater than the sum of the effects of the individual components.

The present combination may advantageously be used to treat and/or prevent the development of a disease, and thus the invention relates to a method for treating and/or preventing the development of a disease in a mammal, and said method comprises administering the present combination to a mammal. In other words, the invention also relates to the use of the combination according to the present invention for the preparation of a composition, preferably a nutritional composition, for the treatment and/or prevention of a disease. In other words, the invention relates to a composition or nutritional composition containing a combination according to the present invention for use in the treatment and/or prevention of a disease. Preferably the mammal is a human, even more preferably the human is a child. Thus, the invention also relates to the use of the combination according to the present invention for the preparation of a composition, preferably a nutritional preparation, for the treatment and/or prevention of the development of a disease in children. In other words, the invention relates to a composition or nutritional composition containing a combination according to the present invention, for use in the treatment and/or prevention of a disease in children.

In the context of the present invention, the child is between 0 and 6 years old, preferably between 0 and 4 years old, preferably between 0 and 2 years old, preferably between 0 and 1 year old.

The invention also relates to a method of providing nutrition to children, including the introduction of the present combination or nutritional composition to the child. In other words, the invention also relates to the use of the combination according to the present invention for the preparation of a nutritional composition for providing nutrition to a child. In other words, the invention relates to a composition or nutritional composition containing a combination according to the present invention for use in providing nutrition to a child.

The present combination can advantageously be used to enhance the Th1 response, increase the Th1/Th2 balance, restore an imbalance in Th1/Th2 responses, maintain a preferred Th1/Th2 balance, and/or treat and prevent the development of diseases that are associated with the Th1/Th2 imbalance. Thus, proposed compositions for, for example, stimulating maturation of the immune system, enhancing resistance to pathogens by stimulating the immune system, and/or supporting the immune system are part of the present invention. In a further embodiment, the present invention provides a method for treating and/or preventing the development of diseases associated with the immune system, comprising administering to the above mammals a composition containing a therapeutically effective amount of the present combination. In a further embodiment, the present invention provides a method for enhancing an immune response in a mammal, comprising administering the present combination to the mammal.

The immune system of a newborn child is characterized by an enhanced Th2 response. As the immune system matures, the Th1 response increases and the Th1/Th2 balance switches to values seen in adults. Thus, the present combination is particularly advantageous for human children. The present invention supports the maturation of the immune system in children. In a further embodiment, the method of the invention relates to administering the present combination to humans 0 to 6 years of age, preferably 0 to 4 years of age, preferably 0 to 2 years of age, more preferably 0 to 1 year of age. In a preferred embodiment, the method relates to stimulating the maturation of the immune system in humans 0 to 6 years of age, preferably 0 to 4 years of age, preferably 0 to 2 years of age, more preferably 0 to 1 year of age.

A composition containing fucosyllactose and betagalactooligosaccharides is even more advantageously used in preterm infants and/or very low or low birth weight infants, since such infants are susceptible and/or prone to viral infections.

A composition containing fucosyllactose and betagalactooligosaccharides is even more preferentially used in children born by caesarean section. Babies born after caesarean section are born in a hospital in an environment that has more pathogens against which antibodies given by the mother to the child are ineffective. Babies born after caesarean section have less optimal colonization and thus are more susceptible to intestinal infections.

Too low a Th1/Th2 balance leads to extreme sensitivity to foreign substances, which can lead to a variety of immunological reactions, such as allergies and related diseases such as atopic dermatitis, asthma, food allergies, allergic rhinitis (eg hay fever). ), dust mite allergy, and other forms of hypersensitivity such as generalized anaphylactic reaction and acute urticaria. Thus, the present combination is particularly useful in treating and/or preventing the development of a disease selected from the group consisting of allergy, food allergy, atopic dermatitis, asthma, allergic rhinitis, dust mite allergy, and urticaria. The present invention improves the balance of Th1/Th2.

An increase in the Th1 response leads to an increase in the response against pathogenic bacteria and/or viruses. Thus, the present combination is suitable for the treatment and/or prevention of infections. This drug can be advantageously used to treat and/or prevent the development of intestinal infections, systemic infections and/or respiratory tract infections.

Since the combination of FL and betagalactooligosaccharides has been found to specifically synergistically enhance the Th1 response, the present invention is particularly suitable for preventing viral infections, more preferably viral infections caused by the orthomyxoviridae family, in particular influenza virus, herpesviridae family, rotavirus, cytomegalovirus, calciviridae family, respiratory syncytial virus, human immunodeficiency virus and/or rhinovirus. The use of the present invention is thus preferred for the prevention and/or treatment of viral infections, more preferably for viral infections, the common cold, influenza, measles, chickenpox, viral diarrhea, viral gastroenteritis, HIV infection and/or viral respiratory tract infection.

It has also been found that the present combination can be suitably used to support vaccination processes, eg enhance the effects of vaccination. The present combination is suitable for maintaining a response to vaccination before, during and/or after vaccination. In particular, the effects of vaccinations against diphtheria-tetanus, whooping cough, polio vaccine, measles/mumps/rubella vaccine, pneumococcal conjugate vaccine, Haemophilus influenzae B conjugate vaccine, hepatitis B, A, chickenpox and/or influenza can be suitably enhanced . Thus, the present combination is advantageously used in the treatment and/or prevention of infections and/or for use in enhancing response to vaccination.

Although the present combination is primarily intended for people suffering from immunodeficiency, in particular for the elderly, suffering from "aging" of the immune system, people with AIDS or infected with HIV, for patients with cancer, in particular for patients with cancer and undergoing chemotherapy, radiotherapy, and patients with cancer cachexia.

The present composition is mainly used for nutrition of the elderly. Older people have a reduced Th1 response. The elderly are particularly susceptible to the complications of a viral infection. In a preferred embodiment, the present invention is used to treat and/or prevent the development of "senescence" of the immune system in the elderly. In one embodiment, the present invention relates to the provision of nutrition to the elderly. An elderly person is considered to be a person aged 55 years or more, in particular aged 65 years or more.

Examples

Example 1

The effect of diets containing 2'-FL and/or betagalactooligosaccharides was tested in mouse models where response to antigen challenge was measured by delayed-type hypersensitivity (DTH) response. This DTH response in the ear after local exposure to the antigen present in the vaccine represents the extent of Th1 cell proliferation. During response to infection and/or vaccination, Th1 cells proliferate in response to antigen exposure. These Th1 cells infiltrate the ear when the ear is subsequently exposed to antigen and cause edema to develop. Infiltration of Th1 cells into the ear takes approximately 24 hours and the onset of edema is thus delayed. The more Th1 cells proliferate during primary vaccination and/or infection, the more DTH is observed in response to antigen exposure.

Materials and methods

6-8 week old female C57BL/6 mice (Charles River) received partially clarified diets based on AIN-93G (Research Diet Service, Wijk bij Duurstede, The Netherlands) including:

1) 1% wt. betagalactooligosaccharide (GOS, source Vivinal GOS, Borculo Domo), fructooligosaccharide (FOS, source RaftilineHP, Orafti) and galacturonic acid oligosaccharide (source AOS) in a ratio of 9:1:1.1. AOS is produced from pectin (Südzucker AG, Mannheim, Germany) with a DP of 1-20. It consists of approximately 75% galacturonic acid oligomers, based on total weight,

2) 1% wt. 2'-fucosyllactose (2'-FL),

3) 1% wt. betagalactooligosaccharides, fructooligosaccharides and galacturonic acid oligosaccharide (AOS) in a ratio of 9:1:1.1 and 1% of the mass. 2'-FL,

4) 0.5% wt. betagalactooligosaccharides, fructooligosaccharides and galacturonic acid oligosaccharide (AOS) in a ratio of 9:1:1.1, and 0.5% of the mass. 2'-FL.

All groups were compared to an unsupplemented control diet (control group and dummy control group). Dietary support began 14 days before the first vaccination and lasted until the end of the experiment, until the 31st day after the first vaccination.

Vaccination experiments were carried out using Influvac (Solvay Pharmaceuticals, Weesp, The Netherlands) from the 2005/2006 season. Mice received a primary vaccination and a booster consisting of a 1:1 subcutaneous (SC) injection of a mixture of vaccine and adjuvant in a total volume of 100 µl. Revaccination was carried out on the 21st day after the primary vaccination. The experiments were completed 10 days after the revaccination. The negative control groups that were included received injections of 1:1 mixture of vaccine and adjuvant in a total volume of 100 μl (dummy control). Ear thickness was measured in duplicate before vaccine exposure and 24 hours after using a digital micrometer (Mitutoyo Digimatic 293561, Veenendaal, The Netherlands). The DTH response was calculated by subtracting the initial ear thickness value from the value obtained 24 hours after exposure.

results

Support with only GOS/FOS/AOS or 2'-FL resulted in increased ear edema. However, the combination of 2'-FL and GOS/FOS/AOS resulted in an even more enhanced DTH response, a Th1 dependent parameter, compared to fed control animals and compared to single components. The combination synergistically enhanced the response to vaccination. See table 1.

Table 1
Effect of 2'-FL and TOS on DTH response Dietary Intervention Ear edema DTH (root mean square error) ∆ DTH µm Relative DTH False control 19.3 (5.5) 0 0 Control 66.3 (6.4) 47.0 1 1% TOS/FOS/AOS 90.8 (7.5) 71.5 1.52 1% 2'-FL 99.7* (10.2) 80.4 1.71 0.5% TOS/FOS/AOS, 0.5% 2'-FL 117.8** (4.4) 98.5 2.10 expected 95.3 76.0 1.62 * denotes p<0.05 compared to control group
** denotes p<0.01 compared to control group and p<0.05 compared to 0.5% GOS/FOS/AOS

Because the effect of DTH response is significantly higher (110%) than DTH responses on diets containing FL (71%) or betagalactooligosaccharides (52%) alone, and also significantly higher than based on the total effect, which can be calculated as 62% th increase in DTH, these results are indicative of the synergistic effect observed with the administration of FL and betagalactooligosaccharides, and preferably fructooligosaccharides and/or uronic acid oligosaccharides, in enhancing the Th1 response.

In summary, these results confirm that oral FL support in combination with betagalactooligosaccharides, more preferably with the additional inclusion of fructooligosaccharides and/or uronic acid oligosaccharides, stimulates the immune response. The immune response is particularly enhanced by the Th1 response and/or the vaccination response. The results are also indicative of an increase in Th1/Th2 balance.

The results of this experiment are indicative that the present invention can advantageously be used to maintain a response to vaccination. The results of this experiment are also indicative in that it can be predominantly used in subjects with a low Th1 response, in particular children. The results of this experiment are also indicative in that it can be advantageously used in subjects with a low Th1 response, in particular in the elderly, suffering from or at risk of "senescence" of the immune system, in patients with HIV, patients with AIDS, and / or in patients with cancer and undergoing chemotherapy, radiation therapy, and patients with cancer cachexia. This model is indicative of basic immunological changes that can be useful in all diseases associated with impaired functioning of the immune system.

Example 2

Immune Boosting Infant Formula containing per 100 ml (13.9 dry weight):

1.4 g proteins (whey and casein),

7.3 g digestible carbohydrates (including lactose),

3.6 g fats (vegetable fat, fish oil),

0.8 g of indigestible oligosaccharides, of which 80 mg of 2'-fucosyllactose and 640 mg of betagalactooligosaccharides and 80 mg of fructooligosaccharides.

Additionally included are: choline, myoinositol, taurine, minerals, trace elements and vitamins as known in the art.

Example 3

Immune Boosting Infant Formula containing per 100 ml (13.9 dry weight):

1.4 g proteins (whey and casein),

7.3 g digestible carbohydrates (including lactose),

3.6 g fats (vegetable fat, fish oil),

0.8 g of indigestible oligosaccharides, of which 40 mg of 2'-fucosyllactose and 760 mg of betagalactooligosaccharides, fructooligosaccharides and uronic acid oligosaccharides in a ratio of 9:1:1.2 by weight.

Additionally included are: choline, myoinositol, taurine, minerals, trace elements and vitamins as known in the art.

Claims (9)

1. Nutritional composition that is not human milk and contains from 0.07 to 1 wt.% 2'-fucosyllactose based on the dry weight of the composition and betagalactooligosaccharides having the structure [galactose] _n -glucose; where n is an integer from 2 to 60, where betagalactooligosaccharides have more than 80% beta-1,4 and beta-1,6 bonds and where the nutritional composition contains proteins, digestible carbohydrates and fats, where proteins provide 5-50%, digestible carbohydrates provide 15-85% and fats provide 5-50% of calories based on total calories. 2. Nutritional composition according to claim 1, which additionally contains fructooligosaccharides and/or uronic acid oligosaccharides. 3. Nutritional composition according to claim 1 or 2, which further comprises fructooligosaccharides and uronic acid oligosaccharides. 4. Nutritional composition according to claim 1 or 2, wherein the betagalactooligosaccharides have more than 90% beta-1,4 and beta-1,6 bonds. 5. Nutritional composition according to claim 2, having a total of 0.25 to 15 wt.% betagalactooligosaccharides, fructooligosaccharides and uronic acid oligosaccharides based on the dry weight of the composition. 6. The use of a nutritional composition according to any one of claims 1 to 5 in preparing a composition for providing nutrition to children. 7. The use of a nutritional composition according to any one of claims 1 to 5 in preparing a composition for providing nutrition to the elderly, cancer patients and/or patients infected with HIV. 8. The use of a nutritional composition according to any one of claims 1 to 5 in the preparation of a composition for the treatment and/or prevention of the development of a disease that can be prevented and/or cured by enhancing the Th1 response. 9. The use of a nutritional composition according to any one of claims 1 to 5 in preparing a composition for enhancing the response to vaccination.