WO2012044155A1

WO2012044155A1 - Fat composition for programming immune response

Info

Publication number: WO2012044155A1
Application number: PCT/NL2010/050633
Authority: WO
Inventors: Johan Garssen; Naomi Van Vlies; Astrid Hogenkamp; Hubertus Josephus Maria Van De Heijning
Original assignee: N.V. Nutricia
Priority date: 2010-09-28
Filing date: 2010-09-28
Publication date: 2012-04-05
Also published as: EP2621291A1; CN103228159A; WO2012044162A1

Abstract

The present invention relates to a method for enhancing vaccination response and/or preventing infections in a human subject later in life, by- administering said human subject a nutritional composition with a specific fat composition having a defined weight ratio of linoleic acid to alpha-linolenic acid when said human subject is an infant with an age between 0 and 36 months.

Description

FAT COMPOSITION FOR PROGRAMMING IMMUNE RESPONSE

FIELD OF THE INVENTION

This invention relates to an infant nutritional composition with a specific fat composition for improving the immune response later in life.

BACKGROUND OF THE INVENTION

Infant feeding practices in the first weeks or months of life influence the susceptibility to diseases later in life. Breast-feeding is the preferred method of feeding infants because of its many health benefits, in particular immune-related benefits. Several studies have shown enhanced vaccination responses in breast-fed compared with non- breast-fed infants. Furthermore, many studies have demonstrated the protective effect of breast-feeding against infections.

Breast-feeding not only reduces the risk of infections during infancy, it also reduces the risk of infections for several years after termination of breast-feeding. Such a long-term protective effect of breast-feeding has been shown for protection in later life against diarrhea, otitis media, respiratory tract infections and Haemophilus influenza type B infection. Although it is not clear which components of breast-milk provide these beneficial health effects later in life, the role of the two essential fatty acids linoleic acid (LA) and alpha-linolenic acid (ALA) and of the long chain polyunsaturated fatty acids (LC-PUFAs) has received a lot of attention.

Not all infants are in the position of being breastfed. It is therefore a continuing aim to provide infant formulas which offer the beneficial functions of human milk such as immune-related benefits, including long-term benefits. WO 2008/056983 discloses the use of a composition containing probiotic bacteria, prebiotics and at least one polyunsaturated fatty acid (PUFA), as a food or therapeutic composition for preventing respiratory diseases in children.

WO 2007/046699 discloses the use of a composition comprising LC-PUFAs and nucleotides for the prevention of infection in an infant delivered via caesarean section. Gibson et al., 2009, Br J Nutr 101 : 1706-1713 show that supplementing infant formula with a probiotic and LC-PUFAs has no effect on the vaccination response in infants of 7 months of age.

Venuta et al., 1996, J Int Med Res 24:325-330 show that recurrent respiratory infections are lowered in children aged between 36 and 49 months receiving LA and ALA as dietary supplements.

Ben et al., 2004, Chin Med J 117: 1268-1270 show that supplementing infant formula with LC-PUFAs, LA and a low amount of ALA has no effect on the incidence of respiratory and intestinal infections in children aged up to 6 months. Bazinet et al., 2004 Immunol Lett 95 : 86-90 reports that in pigs immunized with Mycobacterium tuberculosis, a diet rich in ALA consumed through breast-milk and after weaning enhances the DTH reaction compared to pigs fed a standard commercial pig diet, whereas in non- immunized pigs, the high ALA diet has no effect on the DTH response. None of the above-mentioned compositions are described to program the immune system and to have long-term protective effects against infection later in life when the infant nutrition is no longer consumed.

Das, 2004 Med Sci Monit, 10(5):HY19-25 proposes that perinatal supplementation of omega-3 and omega-6 long-chain polyunsaturated fatty acids (LC-PUFAs) prevents the development of various diseases in adult life.

SUMMARY OF THE INVENTION

The present inventors found in a rodent model that a diet relatively rich in alpha- linolenic acid (ALA) and low in linoleic acid (LA) and with a low LA/ ALA ratio, administered early-in-life to infants via the nursing mother's milk surprisingly increases the vaccination response (T-helper 1 mediated immune response) in this offspring during their later life, indicating a better immune response in later life.

Unexpectedly, the diet not only increased the vaccination response (T-helper 1 mediated immune response) later in life but it also decreased the allergic response later in life (T-helper 2 immune response), indicating a general positive effect on the immune system (both Thl and Th2 responses) later in life.

Because the content of omega-3 and omega-6 fatty acids in the maternal diet is known to be mirrored in the mother's milk composition, the present findings can be applied to the design of an infant nutritional composition. Hence, the present invention relates to an infant nutritional composition with a lipid fraction high in ALA and with a low LA/ ALA ratio. This infant nutritional composition can be advantageously used for the enhancement of the vaccination response and/or for the prevention of infections, wherein the composition is administered early in life during infancy and the advantageous effect is occurring later in life, preferably during childhood and/or adolescence.

DETAILED DESCRIPTION OF THE INVENTION

The invention concerns a method for enhancing vaccination response in a human subject with an age above 36 months, said method comprising administering a lipid component or a composition comprising a lipid component to said human subject when said human subject has an age below 36 months, wherein the lipid component, comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5.

In other words the invention concerns the use of a lipid component or a composition comprising a lipid component, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha- linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5, for the manufacture of a nutritional composition for use in enhancing vaccination response in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

The invention can also be worded as a lipd component or a composition comprising a lipid component, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5 for use in enhancing vaccination response in a human subject with an age above 36 months and the composition is administered to said human subject when said human subject has an age below 36 months. Preferably the composition is a nutritional composition.

The invention also concerns a method for enhancing vaccination response in a human subject with an age above 36 months, said method comprising administering a nutritional composition to said human subject when said human subject has an age below 36 months, wherein the nutritional composition comprises a lipid component, protein component and digestible carbohydrate component, wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60% of the total calories and wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha- lino lenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5.

In other words the invention concerns the use of a composition comprising a lipid component, protein component and digestible carbohydrate component wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60%> of the total calories, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5, for the manufacture of a nutritional composition for use in enhancing vaccination response in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

The invention can also be worded as a composition comprising a lipid component, protein component and digestible carbohydrate component wherein the lipid component provides 35 to 55%> of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha- linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5 for use in enhancing vaccination response in a human subject with an age above 36 months and the composition is administered to said human subject when said human subject has an age below 36 months. Preferably the composition is a nutritional composition.

The invention also concerns a method for preventing infections in a human subject with an age above 36 months, said method comprising administering a lipid component or a composition comprising a lipid component to said human subject when said human subject has an age below 36 months, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5. In other words the invention concerns the use of a lipid component or a composition comprising a lipid component, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha- linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5, for the manufacture of a nutritional composition for use in preventing infections in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

The invention can also be worded as a lipid component or a composition comprising a lipid component, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5 for use in preventing infections in a human subject with an age above 36 months and the composition is administered to said human subject when said human subject has an age below 36 months. Preferably the composition is a nutritional composition. The invention also concerns a method for preventing infections in a human subject with an age above 36 months, said method comprising administering a nutritional composition to said human subject when said human subject has an age below 36 months, wherein the nutritional composition comprises a lipid component, protein component and digestible carbohydrate component, wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60% of the total calories and wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha- linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5.

In other words the invention concerns the use of a composition comprising a lipid component, protein component and digestible carbohydrate component wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60%> of the total calories, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5, for the manufacture of a nutritional composition for use in preventing infections in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

The invention can also be worded as a composition comprising a lipid component, protein component and digestible carbohydrate component wherein the lipid component provides 35 to 55 %> of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60% of the total calories, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha- linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5 for use in preventing infections in a human subject with an age above 36 months and the composition is administered to said human subject when said human subject has an age below 36 months. Preferably the composition is a nutritional composition.

Preferably the present method or use is for enhancing vaccination response and for preventing infections. Lipid component

Omega-3 (referred to as ω^~3, n3 or n-3) fatty acids are unsaturated fatty acids that have in common a final carbon-carbon double bond in the n-3 position, that is the third bond from the methyl end of the fatty acid. In omega-6 (referred to as ω^~6, n6 or n-6) fatty acids the final carbon-carbon double bond is in the n-6 position. Nutritionally important omega-3 fatty acids include alpha-linolenic acid (ALA, 18:3n3), eicosapentaenoic acid (EPA, 20:5n3) and docosahexaenoic acid (DHA, 22:6n3).

The polyunsaturated fatty acids (PUFAs) linoleic acid (LA, 18:2n6, an omega-6 fatty acid) and alpha-linolenic acid (ALA, 18:3n3, an omega-3 fatty acid) are the only two fatty acids that cannot be synthesized in the human body from other substrates, and therefore are called essential fatty acids. They must be present in the diet. They form the starting point for the creation of longer and more unsaturated fatty acids, which are also referred to as long-chain polyunsaturated fatty acids (LC-PUFAs) comprising at least 20 carbon atoms in the fatty acid chain and with 2 or more unsaturated bonds. The omega-3 ALA can be converted into EPA (20:5n3) which can be further converted into DPA (docosapentanenoic acid, 22:5n3), and further into DHA (22:6n3). The omega-6 LA can be converted into arachidonic acid (AA, 20:4n6, an omega-6 fatty acid).

The present inventors have found that a specific composition that has a low LA/ ALA ratio and that is high in ALA has unexpected long-term effects on the immune response in an infant when said composition is administered postnatally to the infant. Particularly the administration of a nutritional composition comprising (i) a LA ALA ratio between 1 and 7.5 and (ii) a high ALA content (>2 wt% based on total fatty acids) resulted in an increased immune response later in life, as observed by an increased Thl response. This finding enables the development of an optimal infant nutritional composition for improving the immune response of the infant later in life. The present composition comprises lipid. ALA should be present in an amount sufficient to enhance Thl response later in life. The present composition therefore preferably comprises at least 2 wt% ALA, more preferably at least 3 wt% ALA, even more preferably at least 4 wt% ALA based on total fatty acids. The present composition preferably comprises less than 50 wt% ALA base on total fatty acids. Based on total dry weight of the composition, the present composition preferably comprises at least 0.50 wt% ALA, more preferably at least 0.75 wt% ALA, even more preferably at least 1 wt% ALA based on total dry weight. The present composition preferably comprises less than 13 wt% ALA based on total dry weight of the composition. When in liquid form, e.g. as ready-to-drink formula, the ALA content is preferably at least 70 mg ALA per 100 ml of the liquid composition, more preferably at least 100 mg ALA per 100 ml of the liquid composition, even more preferably at least 140 mg ALA per 100 ml of the liquid composition. The present composition preferably comprises less than 1750 mg ALA per 100 ml of the liquid composition. LA should be present in a sufficient amount in order to promote a healthy growth and development. The composition therefore comprises less than 50 wt% LA, preferably less than 40 wt% LA, more preferably less than 35 wt% LA, most preferably less than 25 wt% LA based on total fatty acids. The composition preferably comprises at least 2 wt% LA based on total fatty acids. Based on total dry weight of the composition, the present composition comprises less than 13 wt% LA, preferably less than 10.5 wt% LA, more preferably less than 9 wt% LA, most preferably less than 6.5 wt% LA based on total dry weight. The composition preferably comprises at least 0.5 wt% LA based on total dry weight. When in liquid form, e.g. as ready-to-drink formula, the LA content is less than 1750 mg LA per 100 ml of the liquid composition, preferably less than 1400 mg LA per 100 ml of the liquid composition, more preferably less than 1225 mg LA per 100 ml of the liquid composition, most preferably less than 875 mg LA per 100 ml of the liquid composition. The composition preferably comprises at least 70 mg LA per 100 ml of the liquid composition

The weight ratio LA/ALA should be well balanced in order to improve immune response later in life, while at the same time ensuring a normal growth and development. The present composition comprises a weight ratio of LA/ ALA between 1 and 7.5, more preferably between 2 and 6, even more preferably between 3 and 6, even more preferably between 4 and 5.5, most preferably between 4 and 5. The lipid component preferably comprises less than 40 wt% LA based on total fatty acids and a LA/ALA ratio of 2 to 6.

As arachidonic acid (AA) plays an important role in neurological development in infants, the present composition preferably comprises A A. The AA content is preferably at least 0.02 wt%, more preferably at least 0.05 wt%, more preferably at least 0.1 wt%, even more preferably at least 0.2 wt% based on total fatty acids. Since AA, an omega-6 LC-PUFA, may counteract the "programming" effect of the low LA content and low LA/ ALA ratio of the present composition on the immune system, the AA content preferably does not exceed 5 wt%, more preferably does not exceed 1 wt%, more preferably does not exceed 0.5 wt%, even more preferably does not exceed 0.25 wt%, most preferably does not exceed 0.05 wt% based on total fatty acids. Based on total dry weight of the composition, the present composition comprises preferably at least 0.005 wt% AA, more preferably at least 0.01 wt% AA, more preferably at least 0.025 wt% AA based on total dry weight, even more preferably at least 0.05 wt% AA based on total dry weight. The present composition preferably comprises less than 1.3 wt% AA, more preferably less than 0.30 wt% AA, more preferably less than 0.15 wt% AA, even more preferably less than 0.07 wt% AA, most preferably less than 0.015 wt% AA based on total dry weight of the composition. When in liquid form, e.g. as ready-to-drink formula, the AA content is preferably at least 0.70 mg AA per 100 ml of the liquid composition, preferably at least 1.75 mg AA per 100 ml of the liquid composition, more preferably at least 3.50 mg AA per 100 ml of the liquid composition, even more preferably at least 7 mg AA per 100 ml of the liquid composition. The present composition preferably comprises less than 175 mg AA per 100 ml of the liquid composition, more preferably less than 35 mg AA per 100 ml of the liquid composition, more preferably less than 17.5 mg AA per 100 ml of the liquid composition, even more preferably less than 9 mg AA per 100 ml of the liquid composition, most preferably less than 2 mg AA per 100 ml of the liquid composition. The LC-PUFAs, 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 composition contains PUFAs in triglyceride and/or phospholipid form.

The present composition comprises a weight ratio of omega-6 to omega-3 PUFA between 1 and 6, more preferably between 2 and 6, even more preferably between 2 and 5.

Preferably the present composition contains at least one, preferably at least two lipid sources selected from the group consisting of linseed oil (flaxseed oil), rape seed oil (including colza oil, low erucic acid rape seed oil and canola oil), salvia oil, perilla oil, purslane oil, lingonberry oil, sea buckthorn oil, hemp oil, high oleic sunflower oil, high oleic safflower oil, olive oil, marine oils, microbial oils, black currant seed oil, echium oil, butter fat, coconut oil and palm kernel oil. Preferably the present composition contains at least one, preferably at least two lipid sources selected from the group consisting of linseed oil, rapeseed oil, coconut oil, high oleic sunflower oil, butter oil and marine oil. The present composition comprises a lipid component providing 35 to 55 % of the total calories, preferably providing 40 to 50% of the total calories. When in liquid form, e.g. as a ready-to-feed liquid, the composition preferably comprises 2.1 to 6.5 g fat per 100 ml, more preferably 3.0 to 4.0 g perl 00 ml. Based on dry weight the present composition preferably comprises 12.5 to 40 wt% fat, more preferably 19 to 30 wt%.

The term 'lipid' or 'lipid component' as used in the present invention refers to the sum of free fatty acids, triglycerides, diglycerides, monoglycerides, phospholipids, sterols and glycolipids. Nutritional composition

Protein component

The present composition comprises a protein component. The protein used in the nutritional preparation is preferably selected from the group consisting of non-human animal proteins (preferably milk proteins), vegetable proteins (preferably soy protein and/or rice protein), free amino acids and mixtures thereof. The present composition preferably comprises casein, whey, hydrolysed casein and/or hydrolyzed whey protein. Preferably the protein component comprises intact proteins, more preferably intact bovine whey proteins and/or intact bovine casein proteins. In one embodiment, the protein is preferably selected from the group consisting of hydrolysed milk protein, hydrolysed whey protein and hydrolysed casein.

Preferably the present composition comprises sweet whey with a reduced concentration of glycomacropeptide and/or acid whey. Preferably the present composition comprises protein derived from β-casein and/or a-lactalbumin. The present composition preferably comprises casein and whey proteins in a weight ratio casein:whey of 10:90 to 90:10, more preferably 20:80 to 80:20.

The present composition comprises a protein component providing 5 to 15% of the total calories, preferably providing 6 to 12% of the total calories, more preferably providing 6 to 10%> of the total calories. When in liquid form, e.g. as a ready-to-feed liquid, the composition preferably comprises 0.5 to 6.0 g, more preferably 1.0 to 3.0 g, even more preferably 1.0 to 2.5 g protein per 100 ml. Based on dry weight the present composition preferably comprises at least 7.0 wt%, more preferably at least 8.0 wt%, most preferably at least 9 or at least 10 wt% protein based on dry weight of the total composition. Preferably, the present composition comprises at most 40 wt%, more preferably at most 15 wt%, preferably at most 20 wt% of protein based on dry weight of the total composition.

The wt% protein based on dry weight of the present composition can be calculated according to the Kjeldahl-method by measuring total nitrogen and using a conversion factor of 6.38 in case of casein, and a conversion factor of 6.25 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.

Digestible carbohydrate component

The present composition comprises a digestible carbohydrate component preferably providing 30 to 60% of the total calories, more preferably providing 40 to 60% of the total calories, more preferably providing 45 to 55% of the total calories. When in liquid form, e.g. as a ready-to-feed liquid, the composition preferably comprises 3.0 to 30 g digestible carbohydrate per 100 ml, more preferably 6.0 to 20, even more preferably 7.0 to 10.0 g per 100 ml. Based on dry weight the present composition preferably comprises 20 to 80 wt%, more preferably 40 to 65 wt% digestible carbohydrates.

Preferred digestible carbohydrate sources are lactose, glucose, sucrose, fructose, galactose, maltose, starch and maltodextrin. Lactose is the main digestible carbohydrate present in human milk. The present composition preferably comprises lactose.

The present composition preferably comprises digestible carbohydrate, wherein at least 35 wt%, more preferably at least 50 wt%, more preferably at least 75 wt%, even more preferably at least 90 wt%, most preferably at least 95 wt% of the digestible carbohydrate is lactose. Based on dry weight the present composition preferably comprises at least 25 wt% lactose, preferably at least 40 wt%.

The term 'digestible carbohydrate' or 'digestible carbohydrate component' as used in the present invention refers to the sum of digestible carbohydrates.

Non-digestible oligosaccharides

Non-digestible oligosaccharides are present in human milk and are involved in the protection against infection and in stimulating the immune system of the breastfed infants. Therefore, the present composition preferably comprises non-digestible oligosaccharides.

Preferably the present composition comprises non-digestible oligosaccharides with a degree of polymerization (DP) from 2 to 250, more preferably from 3 to 60.

The non-digestible oligosaccharide is preferably selected from the group consisting of fructo-oligosaccharides (FOS) (such as inulin), galacto-oligosaccharides (GOS) (such as transgalacto-oligosaccharides or beta-galacto-oligosaccharides), gluco- oligo saccharides (such as gentio-, nigero- and eye lodextrin-oligo saccharides), arabino-oligosaccharides, mannan-oligosaccharides, xylo-oligosaccharides, fuco- o ligo saccharides, arabino galacto -o ligo saccharides, glucomanno -o ligo saccharides , galactomanno-oligosaccharides, sialic acid comprising oligosaccharides and uronic acid oligosaccharides.

Preferably the present composition comprises fructo-oligosaccharides, galacto- oligosaccharides and/or galacturonic acid oligosaccharides, more preferably galacto- oligosaccharides, most preferably transgalacto-oligosaccharides.

In a preferred embodiment the composition comprises a mixture of transgalacto- oligosaccharides and fructo-oligosaccharides. Preferably the present composition comprises galacto-oligosaccharides with a DP of 2-10 and/or fructo-oligosaccharides with a DP of 2-60. The galacto-oligosaccharide is preferably selected from the group consisting of transgalacto-oligosaccharides, lacto-N-tetraose (LNT), lacto-N- neotetraose (neo-LNT), fucosyl-lactose, fucosylated LNT and fucosylated neo-LNT.

In a particularly preferred embodiment the composition comprises transgalacto- oligosaccharides ([galactose]_n-glucose; wherein n is an integer between 1 and 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). Transgalacto-oligosaccharides (TOS) are for example sold under the trademark Vivinal™ (Borculo Domo Ingredients, Netherlands). Preferably the saccharides of the transgalacto-oligosaccharides are β-linked.

Fructo-oligosaccharide is a non-digestible oligosaccharide comprising a chain of β linked fructose units with a DP or average DP of 2 to 250, more preferably 10 to 100. Fructo-oligosaccharide includes inulin, levan and/or a mixed type of polyfructan. An especially preferred fructo-oligosaccharide is inulin. Fructo-oligosaccharide suitable for use in the compositions is also already commercially available, e.g. Raftiline®HP (Orafti).

Uronic acid oligosaccharides are preferably obtained from pectin degradation. Uronic acid oligosaccharides are preferably galacturonic acid oligosaccharides. Hence the present composition preferably comprises a pectin degradation product with a DP between 2 and 100. Preferably the pectin degradation product is prepared from apple pectin, beet pectin and/or citrus pectin. Preferably the composition comprises transgalacto-oligosaccharide, fructo-oligosaccharide and a pectin degradation product. The weight ratio transgalacto-oligosaccharide : fructo-oligosaccharide : pectin degradation product is preferably (20 to 2) : 1 : (1 to 3), more preferably (12 to 7) : 1 (1 to 2).

GOS, FOS and/or uronic acid oligosaccharides stimulate the immune system of the infant. Hence, the addition of GOS, FOS and/or uronic acid oligosaccharides, preferab ly GO S and FO S , more preferably GO S , FO S and uronic acid oligosaccharides to the composition of the present invention will result in a further improved effect on the immune response.

Preferably, the composition comprises 80 mg to 2 g non-digestible oligosaccharides per 100 ml, more preferably 150 mg to 1.50 g, even more preferably 300 mg to 1 g per 100 ml. Based on dry weight, the composition preferably comprises 0.25 wt% to 20 wt%, more preferably 0.5 wt% to 10 wt%, even more preferably 1.5 wt% to 7.5 wt%.

Formulae

The present composition is preferably particularly suitable for providing the daily nutritional requirements to a human with an age of 36 months or below, particularly a human subject with the age of 24 months or below, even more preferably a human subject with the age of 18 months or below, most preferably a human subject with the age of 12 months or below. Hence, the nutritional composition is used for feeding a human subject. The present composition is preferably enterally administered, more preferably orally.

The present composition is not human breast milk. The present composition comprises a lipid and a protein and a digestible carbohydrate component. The composition of the invention preferably comprises other fractions, such as vitamins, minerals, trace elements and other micronutrients according to international directives for infant formulae and for follow-on formulae.

The present composition comprises a lipid, and a protein and a digestible carbohydrate component wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60%> of the total calories. Preferably the present composition comprises a lipid component providing 40 to 50% of the total calories, a protein component provides 6 to 12% of the total calories and a digestible carbohydrate component provides 40 to 60% of the total calories. The amount of total calories is determined by the sum of calories derived from protein, lipids and digestible carbohydrates. Preferably the lipid component of the present composition comprises less than 40 wt% linoleic acid (LA) based on total fatty acids and comprises at least 3 wt% alpha- lino lenic acid (ALA) based on total fatty acids.

Preferably in the present composition the weight ratio of linoleic acid (LA) to alpha- linolenic acid (ALA) is between 2 and 6. In one embodiment of the present invention the lipid component preferably provides 35 to 50%, preferably 40 to 50% of the total calories, the protein component preferably provides 6 to 12%, preferably 6 to 10% of the total calories, and the digestible carbohydrate component preferably provides 40 to 60% preferably 45 to 55% of the total calories, wherein the lipid component comprises less than 40 wt% linoleic acid (LA), preferably less than 35 wt% LA, more preferably less than 25 wt% LA based on total fatty acids, and comprises at least 3 wt% alpha-linolenic acid (ALA), preferably at least 4 wt% ALA and wherein the weight ratio of LA to ALA is between 2 and 6, preferably between 3 and 6, more preferably between 4 and 5.5.

In order to meet the caloric requirements of the infant, the composition preferably comprises 50 to 200 kcal/100 ml liquid, more preferably 60 to 90 kcal/100 ml liquid, even more preferably 60 to 75 kcal/100 ml liquid. This caloric density ensures an optimal ratio between water and calorie consumption. The osmolarity of the present composition is preferably between 150 and 420 mOsmol/1, more preferably 260 to 320 mOsmol/1. The low osmolarity aims to reduce the gastrointestinal stress. Preferably the composition is in a liquid form. Preferably the liquid composition has a viscosity below 35 mPa.s, more preferably below 6 mPa.s as measured in a Brookfield viscometer at 20°C at a shear rate of 100 s^"1. When the composition is in a liquid form, the preferred volume administered on a daily basis is in the range of about 80 to 2500 ml, more preferably about 200 to 1200 ml per day. Preferably, the number of feedings per day is between 1 and 10, preferably between 3 and 8. In one embodiment the composition is administered daily for a period of at least 4 weeks, preferably for a period of at least 8 weeks, more preferably for a period of at least 12 weeks, in a liquid form wherein the total volume administered daily is between 200 ml and 1200 ml and wherein the number of feedings per day is between 1 and 10. Preferably the period of at least 4 weeks, or at least 8 weeks, or at least 12 weeks is in the first 24 months of life, more preferably in the first 12 months of life, most preferably in the first 6 months of life of an infant.

In one embodiment the composition is a powder suitable for making a liquid composition after reconstitution with an aqueous solution, preferably with water. Preferably the composition is a powder to be reconstituted with water.

Applications

The present inventors found in an animal study that postnatal administration of a nutritional composition comprising the present lipid component has a long-term impact on the development of the immune system and has a long-term effect on the susceptibility to immune diseases in the offspring. The inventors have recognized an unexpected long-term effect of dietary PUFAs on the immune response when said PUFAs are administered postnatally. Hence, it is important that the present nutritional composition is administered to an infant in the earliest period of life after birth. It is believed that the first weeks or months of life are optimal for establishing long-term "programming" effects of the infant nutrition on the development of diseases later in life. This "programming" effect is formally defined as an "effect that occur when a stimulus or insult operating at a critical or sensitive period of development could result in a long-standing or life-long effect on the structure or function of the organism" (Lucas, A. 2000, Am J Clin Nutr, 71 :602). The important underlying idea in this definition is that the programming event can only occur during a specific window of sensitivity. It is believed that the younger an infant is when receiving the present composition, the more efficient are the programming effects on the immune system which is still under development in such young infant.

In one embodiment of the use according to the present invention, the nutritional composition is advantageously administered to a human subject with an age of 36 months or below, preferably with an age of 24 months or below, more preferably with an age of 12 months or below, most preferably with an age of 6 months or below.

The present inventors found in an animal study that postnatal administration of a nutritional composition comprising the present lipid component up to 3 weeks after birth results in an increase in Thl response after Influenza vaccination in the offspring, the Thl response being measured 11 weeks after birth. Importantly, 3 weeks after birth the fatty acid composition of the offspring's red blood cells mirrored the maternal diet, whereas at the time of vaccination (7 weeks after birth) the red blood cells fatty acid composition of the offspring did not reflect anymore the maternal diet, showing that the differences in vaccination response at 1 1 weeks are true programming effects, and are not due to residual effect from the maternal diet.

The present invention particularly aims to enhance vaccination response, such effect occurring in the human subject at a time when the composition is no longer administered to said human subject. The term "vaccination response" refers to the response of the immune system (immune response) to a vaccine, i.e. a weakened or killed pathogen. The term "vaccination" refers to immunization, the process of stimulating an immune response via use of a weakened or killed infectious agent. Herein, "vaccination" also refers to inoculation, the process of stimulating an immune response via the use of unweakened live pathogens. The present combination can suitably be used to support vaccination processes, e.g. enhance the effects of a vaccination process. The present combination is suitable for supporting vaccination response before, during and/or after vaccination. Particularly the effects of vaccinations for diptheria-tetanus, pertussis, polio vaccine, measles/mumps/rubella, pneumococcal conjugate, haemophilus B conjugate, hepatitis B, hepatitis A, varicella, and/or influenza can suitably be enhanced.

The present composition was found to enhance the DTH response, which is indicative of an enhanced Thl response. An increased Thl response leads to an increase in the response against pathogenic bacteria and/or viruses. Hence, the present composition is suitable for the prevention of infections . The present preparation can be advantageously used for the prevention of intestinal infections and/or respiratory tract infections and/or ear infections and/or urinary tract infections. The present invention particularly aims to prevent infections, such preventive effect occurring in the human subj ect at a time when the composition is no longer administered to said human subject. Herein, the term 'infection' refers to a disease or pathological state caused by the invasion of the body by pathogenic microorganisms. Pathogenic microorganisms, also called pathogens, include viruses, bacteria, fungi, protozoan parasites, macroscopic helminths (worm) parasites, prions and viroids. The most frequent infections in infants (aged 0-3 years) and children (age 3-12 years) are gastrointestinal tract infections, among which diarrhea, respiratory tract infections, ear infections (otitis media) and urinary tract infections. Since the present composition was found to enhance the Thl response, the present invention is particularly suitable to prevent viral infections, more preferably viral infections caused by orthomyxoviridae, in particular the influenza virus, herpesviridae, rotavirus, cytomegalovirus, caliciviridae, respiratory syncytial virus, human imunodeficiency virus and/or rhinovirus. The use of the present invention is therefore preferably for preventing viral infections, more preferably the viral infections common cold, flu, measles, chicken pox, viral diarrhoea, viral gastroenteritis, and/or viral respiratory tract infections.

In one embodiment the present invention aims to enhance the vaccination response and to prevent infections, such effects occurring in the human subject at a time when the composition is no longer administered to said human subject.

In one further embodiment the present invention aims to enhance the vaccination response and/or to prevent infections and to prevent allergy and/or atopic diseases, such as asthma and/or atopic dermatitis, such effects occurring in the human subject at a time when the composition is no longer administered to said human subject. In one further embodiment the present invention aims to enhance the vaccination response and/or to prevent infections, such effects occurring later in life in the human subject with an age above 36 months, preferably with an age above 5 years, more preferably with an age above 8 years, wherein the composition is no longer administered to said human subject. With "later in life" is meant an age exceeding the age at which the nutritional composition is ingested, preferably exceeding said age with at least one year, preferably with 2 years, more preferably with 5 years. 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".

EXAMPLES

Example 1: Programming effect of maternal dietary PUFAs on offspring vaccination response

Methods: Twelve week old C57B1/6 mice were put on the standard rodent diet (control diet, AIN93G, see Table 1). After one week, female mice were mated and randomised to one out of seven groups fed either the control diet (C) during pregnancy and lactation, a high CI 8:3 n-3 (a-linolenic acid, ALA) diet (diet 1) fed either during pregnancy and lactation (1PL) or during lactation only (1L), or a high CI 8:2 n-6 (linoleic acid, LA) diet (diet 2) fed either during pregnancy and lactation (2PL) or during lactation only (2L). Dams fed diet 1 or diet 2 during lactation (1L or 2L) were fed the control diet before giving birth. All diets consisted of 20% casein, 37% cornstarch, 13% maltodextrin, 10% dextrose, 5% Vitacel L600-20 (fiber), 5% mixture of vitamins and minerals and 10% fat. The fatty acid composition of the different diets is listed in Table 1. At weaning (3 weeks after birth), blood samples were taken from both dams and male pups. Pups were fed a western style diet (Table 1) until the end of the experiment. Male mice were vaccinated at 7 and at 9 weeks of age (4 and 6 weeks after weaning respectively) by a subcutaneous (s.c.) injection of 100 μΐ Influvac 2008/2009 (Solvay Pharma B.V., Weesp, The Netherlands). Blood samples were taken from the pups at the time of the first vaccination (7 weeks of age). Delayed-type hypersensitivity responses (DTH) were induced at the age of 1 1 weeks by s.c. injection of 25 μΐ Influvac into the ear pinna of one ear, whilst 25 μΐ of PBS was injected in the other ear. Ear thickness, as readout for T-helper-1 dependent cellular immunity, was measured in duplicate before antigen challenge and 24 h afterwards, with a digital micrometer (Mitutoyo Digimatic 293561 ; Veenendaal, The Netherlands). The DTH response was calculated by subtracting the basal ear thickness from the value at 24 h after challenge, correcting for the ear swelling that occurred as a result of s.c. injection of PBS. Afterwards, mice were euthanized by intraperitoneal injection of 1 ml 10 % urethane. Blood was collected via cardiac puncture into EDTA tubes and stored on ice. To analyze the fatty acid composition of red blood cells (RBC or erythrocytes) of dams and pups, blood samples taken at weaning, at the time of vaccination and at the end of the experiment were centrifuged at 14000 rpm for five minutes, and plasma was removed. Erythrocytes were washed twice with PBS containing 5 mM EDTA. They were resuspended in an equal volume of PBS-EDTA and stored at -80 °C until analysis. Erythrocyte lipids were extracted as described previously (Bligh, 1959) and the fatty acid profile was analysed using HPLC.

Table 1. Fatty acid composition of the different diets (g/100 g total fatty acids)

Results: Red blood cell (RBC) fatty acid composition in dams was measured at weaning (Table 2). RBC fatty acid composition is considered to reflect the pattern of intake of fatty acids (Katan, 1997). Dams in group IPL had significantly higher levels of CI 8:3 n-3 (a-linolenic acid, ALA) and its derivative C20:5 n-3 (eicosapentenoic acid, EPA) in their RBCs compared to all other dietary groups except for 1L (Table 2). The C22:6 n-3 (docosahexaenoic acid, DHA) levels in the IPL dams differed only from the dams in 2PL group. No significant differences were found for other fatty acids. Table 2. Percentages of PUFAs in maternal red blood cells at time of weaning

Mice were fed the control diet (C), diet 1 during pregnancy and lactation (IPL), diet 1 during lactation (1L), diet 2 during pregnancy and lactation (2PL) or diet 2 during lactation (2L). Values are given as mean ± SEM. Groups which do not share a similar letter have a significantly different percentage of that PUFA, p<0.05. The RBC fatty acid composition in pups was measured at the time of weaning (Table 3). The levels of ALA and EPA (a derivative of ALA) in the RBC of all pups reflected that of their mothers RBC and reflected the maternal diet during lactation. The pups from 1L and IPL dams (diet high in ALA) had higher levels of ALA and of its derivative EPA compared to the other groups. AA levels in pups also reflected the maternal diet: 1L and IPL pups (diet low in LA, from which AA is derived) had lower AA levels compared to the other groups. Control pups had lower DHA levels compared to all other dietary groups. No differences between groups were found in linoleic acid levels (results not shown).

At the time of vaccination (7 weeks of age), no differences were found in the RBC fatty acid composition of the pups from the different groups (data not shown).

At the end of the experiment, after 8 weeks on the Western-style diet, there was no longer a difference in the RBC ALA and AA levels in the offspring of the different groups, showing that the direct diet effect on the content of these FA measured at weaning was attenuated 8 weeks later. The only difference found was the increased levels of EPA and/or DHA in the offspring of dams which were fed diet 1 or diet 2 compared to controls (Results not shown). Table 3 : RBC fatty acid composition in pups at the time of weaning

Red blood cell PUFA content (%) at weaning in offspring of mice fed the control diet (C), diet 1 during pregnancy and lactation (1PL), diet 1 during lactation (1L), diet 2 during pregnancy and lactation (2PL) or diet 2 during lactation (2L). Values are given as mean ± SEM. Groups which do not share a similar letter have a significantly different PUFA level, p<0.05.

At the end of the experiment, offspring of dams fed diet 1 during lactation (1L) showed an enhanced DTH response against the vaccine, compared to the control group (Table 4) indicating improved cellular immunity to viral antigens. Thus, a higher intake of 18:3 n-3 (ALA) during the lactation period (1L) has a prominent effect on the offspring's immune response at a later time resulting in enhanced T- helper 1 mediated response. This indicates that the maternal diet with a low LA content, a high ALA content and a low LA/ ALA ratio, influences the immune response in the offspring so that in adulthood the immune response is improved. Table 4: Delayed type hypersensitivity response

T-helper 1 mediated delayed type hypersensitivity response (DTH) in offspring of mice fed the control diet (C), diet 1 during pregnancy and lactation (1PL), diet 1 during lactation (1L), diet 2 during pregnancy and lactation (2PL) or diet 2 during lactation (2L). Values are given as mean ± SEM. Groups which do not share a similar letter have a significantly different ear swelling response, p<0.05. Example 2: Programming effect of maternal dietary PUFAs on allergic response Methods: The composition of the diets and the feeding schedule are the same as described in Example 1. Twelve week old BALB/C mice were put on the standard rodent diet (control diet, AIN93G, see Table 1). After one week, female mice were mated and randomised to one of the diet groups. At weaning (3 weeks after birth), blood samples were taken from both dams and male pups, which were fed a western style diet (Table 1) until the end of the experiment. Male mice were sensitised to ovalbumin at 6 and at 7 weeks of age (3 and 4 weeks after weaning respectively) by intraperitoneal injection of 10 μg ovalbumin adsorbed into 22.5 mg aluminium hydroxide in 100 μΐ saline. One day before the first sensitization a blood sample was taken. Acute allergic skin reaction (ASR) was measured at 9 weeks of age by s.c. injection of 1 pg ovalbumin in 25 μΐ PBS into the ear pinna of one ear. The other ear pinna was injected with 25 μΐ PBS as a control. Ear thickness, as readout for a T- helper 2 mediated allergic reaction, was measured in duplicate before antigen challenge and 1 hr afterwards, using a digital micrometer (Mitutoyo Digimatic 293561; Veenendaal, The Netherlands). The ASR was calculated by subtracting the basal ear thickness from the value at 1 hr after challenge, correcting for the ear swelling that occurred in the PBS injected ear. At the age of 11 weeks, mice were euthanized as described in example 1. Blood collection, RBC isolation, lipid extraction thereof and analysis of the RBC fatty acid composition were all performed as described in Example 1.

Results: The RBC fatty acid composition in dams was measured at weaning and was similar to that described in Example 1. The RBC fatty acid composition in pups was measured at the time of weaning (Table 5). The levels of ALA and EPA (a derivative of ALA) in the RBC of all pups reflected that of their mothers RBC and reflected the maternal diet during lactation. The pups from 1L and 1PL dams (diet high in ALA) had higher levels of ALA and of its derivative EPA compared to the other groups. There was no difference in AA between groups at weaning. The levels of DHA (a derivative of ALA) mirrored the levels of ALA in the maternal diet: control pups and the diet 2 pups had similar DHA levels, which were lower than the levels of the diet 1 pups (Table 5). No differences between groups were found in linoleic acid levels (results not shown). Table 5 : RBC fatty acid composition in pups at the time of weaning

Red blood cell PUFA content at weaning in offspring of mice fed the control diet (C), diet 1 during pregnancy and lactation (IPL), diet 1 during lactation (1L), diet 2 during pregnancy and lactation (2PL) or diet 2 during lactation (2L). Values are given as mean ± SEM. Groups which do not share a similar letter have a significantly different PUFA level, p<0.05.

Before the first sensitization with ovalbumin (6 weeks of age, 3 weeks after weaning), the 1L and IPL mice had no longer a higher ALA content as measured at weaning. They had somewhat higher EPA and DHA levels compared to the other groups (results not shown). No significant differences were found in the AA and LA levels between the different groups (results not shown).

All PUFA diets diminished the T-helper-2 dependent acute allergic skin response (ASR), but the extent of the effect was highly dependent on the feeding period and was not similar between the two diets (Table 6). Offspring of dams fed diet 1 during lactation (1L) showed the most prominent attenuation of the ASR. Thus, a higher intake of 18:3 n-3 during the lactation period (1L) has a prominent effect on the offspring's immune response at a later time resulting in a lower T-helper 2 mediated response.

Table 6: Acute skin response

T-helper 2 mediated acute skin response (ASR) in offspring of mice fed the control diet (C), diet 1 during pregnancy and lactation (IPL), diet 1 during lactation (1L), diet 2 during pregnancy and lactation (2PL) or diet 2 during lactation (2L). Values are given as mean ± SEM. Groups which do not share a similar letter have a significantly different ear swelling response, p<0.05. Example 3: Infant nutrition

Infant nutrition comprising a lipid component providing 48% of the total calories, a protein component providing 8% of the total calories and a digestible carbohydrate component providing 44% of the total calories; (i) the lipid component comprising based on total fatty acids: 13.3 wt% LA; 2.4 wt% ALA; 0.2 wt% DHA; the LA/ ALA ratio is 5.7; (ii) the digestible carbohydrate component comprising 51 gram lactose/100 gram powder; 0.36 g galacto-oligosaccharides with DP 2-6 and 0.04 g fructo -oligosaccharides with DP 7-60; (iii) the protein component comprising cow milk protein. The infant nutrition comprises GOS and FOS in a ratio of 9: 1. The label of the package of this infant nutrition indicates that the nutrition enhances vaccination response later in life and/or prevents infections later in life and/or prevents allergy later in life and/or prevents atopic diseases later in life.

Claims

Use of a composition comprising a lipid component, protein component and digestible carbohydrate component wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60%> of the total calories, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5, for the manufacture of a nutritional composition for use in enhancing vaccination response in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

Use of a composition comprising a lipid component, protein component and digestible carbohydrate component wherein the lipid component provides 35 to 55% of the total calories, the protein component provides 5 to 15% of the total calories and the digestible carbohydrate component provides 30 to 60%> of the total calories, wherein the lipid component comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5, for the manufacture of a nutritional composition for use in preventing infections in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

Use according to claim 1 or 2 for enhancing vaccination response or preventing infections in a human subject with an age of 5 years and above, preferably with an age of 8 years and above.

Use according to any one of the preceding claims wherein the nutritional composition is administered to an infant with an age of 12 months and below.

5. Use according to any one of the preceding claims wherein the nutritional composition is for further use in preventing allergy and/or atopic diseases when said human subject has an age above 36 months.

6. Use according to any one of the preceding claims wherein the composition is administered daily for a period of at least 4 weeks in a liquid form wherein the total volume administered daily is between 200 ml and 1200 ml and wherein the number of feedings per day is between 1 and 10.

7. Use according to any one of the preceding claims wherein the lipid component provides 35 to 50% of the total calories, the protein component provides 6 to 12% of the total calories and the digestible carbohydrate component provides 40 to 60% of the total calories.

8. Use according to any one of the preceding claims wherein the lipid component comprises less than 40 wt% linoleic acid (LA) based on total fatty acids and comprises at least 3 wt% alpha-linolenic acid (ALA) based on total fatty acids.

9. Use according to any one of the preceding claims wherein the weight ratio of linoleic acid (LA) to alpha-linolenic acid (ALA) is between 2 and 6.

10. Use according to any one of the preceding claims wherein the weight ratio of omega-6 to omega-3 poly-unsaturated fatty acids (PUFA) is between 1 and 6

11. Use according to any one of the preceding claims wherein the weight ratio of omega-6 to omega-3 poly-unsaturated fatty acids (PUFA) is between 2 and 5.

12. Use of a lipid component that comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5 for the manufacture of a nutritional composition for use in enhancing vaccination response in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.

13. Use of a lipid component that comprises less than 50 wt% linoleic acid (LA) based on total fatty acids, and comprises at least 2 wt% alpha-linolenic acid (ALA) based on total fatty acids, and wherein the weight ratio of LA to ALA is between 1 and 7.5 for the manufacture of a nutritional composition for use in preventing infections in a human subject with an age above 36 months wherein the nutritional composition is administered to said human subject when said human subject has an age below 36 months.