U.S. patent application number 13/383822 was filed with the patent office on 2012-07-12 for fucosyllactose as breast milk identical non-digestible oligosaccharide with new functional benefit.
This patent application is currently assigned to N.V. NUTRICIA. Invention is credited to Sophie Drouillard, Johan Garssen, Alma Jildou Nauta, Eric Samain, Bernd Stahl.
Application Number | 20120177691 13/383822 |
Document ID | / |
Family ID | 41016877 |
Filed Date | 2012-07-12 |
United States Patent
Application |
20120177691 |
Kind Code |
A1 |
Stahl; Bernd ; et
al. |
July 12, 2012 |
FUCOSYLLACTOSE AS BREAST MILK IDENTICAL NON-DIGESTIBLE
OLIGOSACCHARIDE WITH NEW FUNCTIONAL BENEFIT
Abstract
The invention concerns nutritional compositions with
fucosyllactose for use in stimulation of NK cells. The composition
is suitable for infants.
Inventors: |
Stahl; Bernd;
(Friedrichsdorf, DE) ; Nauta; Alma Jildou;
(Driebergen, NL) ; Garssen; Johan; (Nieuwegein,
NL) ; Samain; Eric; (Gieres, FR) ; Drouillard;
Sophie; (Claix, FR) |
Assignee: |
N.V. NUTRICIA
|
Family ID: |
41016877 |
Appl. No.: |
13/383822 |
Filed: |
July 12, 2010 |
PCT Filed: |
July 12, 2010 |
PCT NO: |
PCT/NL10/50447 |
371 Date: |
March 28, 2012 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
61256453 |
Oct 30, 2009 |
|
|
|
Current U.S.
Class: |
424/278.1 ;
514/61; 536/123.1 |
Current CPC
Class: |
A23L 33/40 20160801;
A23L 29/30 20160801; A23L 33/10 20160801; A61P 37/04 20180101; A23L
33/125 20160801; A61K 2039/572 20130101; A61K 31/7032 20130101;
A61K 31/702 20130101; A23V 2002/00 20130101; A61K 39/39 20130101;
A61P 31/12 20180101; A61K 2039/55 20130101; A23L 5/00 20160801;
A61K 31/702 20130101; A61K 2300/00 20130101; A23V 2002/00 20130101;
A23V 2250/28 20130101; A23V 2200/308 20130101; A23V 2200/324
20130101; A61K 31/702 20130101; A61K 2300/00 20130101 |
Class at
Publication: |
424/278.1 ;
514/61; 536/123.1 |
International
Class: |
A61K 31/702 20060101
A61K031/702; A61P 31/18 20060101 A61P031/18; A61P 31/16 20060101
A61P031/16; C07H 3/06 20060101 C07H003/06; A61P 31/14 20060101
A61P031/14 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 15, 2009 |
EP |
09165485.5 |
Claims
1-9. (canceled)
10. A method for treating and/or preventing viral infections caused
by orthomyxoviridae, comprising administering to a mammal in need
thereof an enteral composition comprising 2'-fucosyllactose,
wherein the composition is not human milk.
11. The method according to claim 10, wherein the orthomyxoviridae
comprise influenza virus, herpesviridae, rotavirus,
cytomegalovirus, respiratory syncytial virus, human imunodeficiency
virus and/or rhinovirus.
12. The method according to claim 10, wherein the mammal suffers
from common cold, flu, measles, chicken pox, viral
gastro-enteritis, HIV infection and/or a viral respiratory tract
infection.
13. The method according to claim 10, wherein the mammal is an
infant, HIV patient, elderly and/or oncology patient.
14. The method according to claim 10, wherein the mammal is a
human.
15. The method according to claim 14, wherein the mammal is a human
infant.
16. The method according to claim 13, wherein the human is a HIV
patient, an elderly person and/or an oncology patient.
17. The method according to claim 10, wherein the composition
further comprises at least one of beta-galacto-oligosaccharides,
fructo-oligosaccharides and uronic acid oligosaccharides.
18. The method according to claim 10, wherein the composition
comprises 0.07 to 1 wt % fucosyllactose, based on dry weight of the
composition.
19. The method according to claim 10, wherein the composition
further comprises, based on total energy of the composition, 5 to
50% protein, 15 to 85% digestible carbohydrates and 5 to 50%
fat.
20. A method for enhancing a vaccination response, comprising
administering to a mammal an enteral composition comprising
2'-fucosyllactose.
21. The method according to claim 20, wherein the response to
vaccination with a viral antigen is enhanced.
22. An infant formula comprising 2'-fucosyllactose.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to infant nutrition with
non-digestible oligosaccharides, in particular to the use thereof
for stimulating the immune system.
BACKGROUND OF THE INVENTION
[0002] Human milk fed infants have a lower incidence of infections,
including viral infections, than formula fed infants. Many
components in human milk, including immunoglobulins (such as IgA),
interleukin (IL)-1, IL-6, IL-8, IL-10, interferon-.gamma.
(IFN-.gamma.), immunocompetent cells, transforming growth factor-13
(TGF-13), lactoferrin, nucleotides, and non-digestible
oligosaccharides (NDO) are thought to be involved in protection
against infection with enteric or respiratory pathogens. TGF-.beta.
and dietary nucleotides were found to be components which may be
responsible for increase in natural killer cell activity.
[0003] NDO are a major constituent of human milk and are a major
element of the innate immune system of human milk. Human NDO
promote the growth of a beneficial microbiota dominated by
bifidobacteria and lactobacilli. Some human NDO are also known to
be able to prevent directly the adhesion of pathogens and
toxins.
[0004] Human milk is the preferred food for infants. However, it is
not always possible or desirable to breast feed an infant. In such
cases infant formulae or follow on formulae are a good alternative.
These formulae should have an optimal composition in order to mimic
the beneficial effects of breast milk as close as possible.
[0005] WO 2007/067053 discloses infant formula comprising the
plant-derived prebiotics inulin and galacturonic acid
oligosaccharide and the from lactose synthesized prebiotic
transgalacto-oligosaccharide to reduce infections.
[0006] WO 2007/010084 discloses mannan-poly- and oligosaccharides
for immune stimulation.
[0007] U.S. Pat. No. 6,576,251 discloses a carbohydrate mixture for
dietetic foods administered by the enteral or parenteral route
consisting of (a) monosaccharide(s), (b) oligosaccharide(s) (at
most hexasaccharides) and (c) polysaccharide(s) (at least
heptasaccharides), where the mixing ratio a, b, c, in respect of
weight, is: alpha=1, b=40 to 1000, and c=1 to 50, and containing at
least 1 weight percent of fucose occurring either freely and/or
bound to an oligosaccharide and/or a polysaccharide. The
carbohydrate mixture is said to have both a nutritional and a
biological effect which is considerably greater than the
corresponding action of the individual constituents.
[0008] EP 1 629 850 provides a method and composition for the
treatment and/or prevention of respiratory tract infection and/or
respiratory tract infection disease, said method comprising orally
administering a composition to a mammal, said composition
comprising a galactose containing indigestible oligosaccharide and
at least 5 wt.% digestible galactose saccharide.
[0009] EP 2 072 052 relates to a composition suitable for use in
the prevention of opportunistic infections in immune-compromised
individuals comprising a probiotic and a fucosylated
oligosaccharide selected from the group comprising
2'-fucosyllactose, 3'fucosyllactose, difucosyllactose,
lacto-N-fucopentaose, lacto-N-fucohexaose, fucosyllacto-N-hexaose
and fucosyllacto-N-neohexaose. The document further discloses the
use of such a composition in the prevention of opportunistic
infections in immune-compromised individuals.
SUMMARY OF THE INVENTION
[0010] Human milk differs from milk from domestic animals in that
it comprises more NDO and in that the NDO are structurally
different. The group of human NDO is very complex, since it
represents a heterogenic group of more than 130 different compounds
with diverse sugar composition. Because of their complex and
polymorphic structure, large-scale synthesis is complicated. It is
therefore not yet technically and economically feasible to prepare
infant nutrition with an NDO composition identical to human
milk.
[0011] Recently, new techniques have become available to chemically
synthesise specific types of NDO identical to specific human NDO,
thereby offering the opportunity to test the immunomodulatory
capacity of specific human NDO in in vitro and in vivo assays.
[0012] The inventors unexpectedly have found that fucosyllactose
(FL), an oligosaccharide abundantly present in human breast milk
and with a relatively simple structure, specifically increases the
number and thereby the activity of Natural Killer (NK) cells. NK
cells play an important role in the natural defence against viral
infections and tumour cells. The finding which specific
oligosaccharide is responsible for increasing NK cell activity now
enables the design of nutritional compositions comprising FL, more
particularly 2'-FL, for the use of increasing NK cells and/or NK
cell activity.
DETAILED DESCRIPTION
[0013] The present invention thus concerns a method for stimulating
NK cell activity and/or NK cell proliferation in a subject, said
method comprising administering a composition comprising
fucosyllactose to said subject, said composition not being human
milk. In one embodiment the present method is a non-medical method
for stimulating NK cell activity and/or NK cell proliferation in a
subject. In one embodiment the present method is for treating
and/or preventing viral infections in a subject.
[0014] The present invention also concerns a method for treating
and/or preventing viral infections in a subject, said method
comprising administering a composition comprising fucosyllactose to
said subject, said composition not being human milk.
[0015] The invention can also be worded as the use of
fucosyllactose in the manufacture of a composition for stimulating
NK cell activity and/or NK cell proliferation, said composition not
being human milk. In one embodiment the composition is for treating
and/or preventing viral infections.
[0016] The invention can also be worded as the use of
fucosyllactose in the manufacture of a composition for treating
and/or preventing viral infections, said composition not being
human milk.
[0017] The invention can also be worded as a composition comprising
fucosyllactose for stimulating, in particular for use in
stimulating, NK cell activity and/or NK cell proliferation, said
composition not being human milk. In one embodiment the composition
is for use in treating and/or preventing, viral infections.
[0018] The invention can also be worded as a composition comprising
fucosyllactose for treating and/or preventing, in particular for
use in treating and/or preventing, viral infections, said
composition not being human milk.
[0019] The invention also concerns a method for treating and/or
preventing infections, by stimulating natural killer (NK) cell
activity and/or NK cell proliferation, said method comprising
administering a composition comprising fucosyllactose to said
subject, said composition not being human milk.
[0020] The invention can also be worded as the use of
fucosyllactose in the manufacture of an enteral composition for
treating and/or preventing infections, by stimulating natural
killer (NK) cell activity and/or NK cell proliferation, said
composition not being human milk.
[0021] The invention can also be worded as a composition comprising
fucosyllactose for treating and/or preventing infections, in
particular for use in and/or preventing infections, by stimulating
natural killer (NK) cell activity and/or NK cell proliferation,
said composition not being human milk.
[0022] The invention also concerns a method for enhancing
vaccination response, said method comprising administering a
composition comprising fucosyllactose to said subject, said
composition not being human milk. In one embodiment the method is
for enhancing vaccination response to vaccination with viral
antigens.
[0023] The invention can also be worded as the use of
fucosyllactose in the manufacture of an enteral composition for
enhancing vaccination response. In one embodiment the composition
is for enhancing vaccination response to vaccination with viral
antigens.
[0024] The invention can also be worded as a composition comprising
fucosyllactose for enhancing, in particular for use in enhancing,
vaccination response. In one embodiment the composition is for
enhancing vaccination response to vaccination with viral
antigens.
[0025] The composition that is administered according to the
present method, or that is used according to the present invention,
is preferably enterally administered, more preferably orally. Or in
other words the composition is preferably for enteral, preferably
oral administration or in other words the composition is an
enteral, preferably oral, composition.
[0026] Fucosyllactose
[0027] The present composition comprises fucosyllactose.
Fucosyllactose (FL) is a non-digestible oligosaccharide present in
human milk. It is not present in bovine milk. It consists of three
monose units, fucose, galactose and glucose linked together.
Galactose linked to glucose via a beta 1.fwdarw.4 linkage is called
lactose. A fucose unit is linked to a galactose unit of a lactose
via an alpha 1,2 linkage (2'-fucosyllactose, 2'-FL) or to the
glucose unit of lactose via an alpha 1,3 linkage (3-fucosyllactose,
3-FL). The present composition preferably comprises 2'-FL.
[0028] 2'-FL, preferably
.alpha.-L-Fuc-(1.fwdarw.2)-.beta.-D-Gal-(1.fwdarw.4)-D-Glc, and
3-FL, preferably
.alpha.-L-Fuc-(1.fwdarw.3)-[.beta.-D-Gal-(1.fwdarw.4)]-D-Glc), are
commercially available for instance 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 produced by genetically modified micro-organisms,
for example as described in Albermann et al, 2001, Carbohydrate
Res. 334:97-103.
[0029] Preferably, the composition comprises 1 mg to 3 g
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
composition preferably comprises 0.007 wt % to 20 wt %
fucosyllactose, more preferably 0.07 wt % to 10 wt %, even more
preferably 0.15 wt % to 1 wt %. A lower amount of fucosyllactose
will be less effective in increasing NK cells and/or increasing NK
cell activity, whereas a too high amount will result in unnecessary
high costs of the product.
[0030] Non-Digestible Oligosaccharides Other than FL
[0031] The present composition preferably comprises non-digestible
oligosaccharides (NDO) other than FL. Preferably the NDO other than
FL stimulate the growth of bifidobacteria and/or lactobacilli, more
preferably bifidobacteria. An increased content of bifidobacteria
and/or lactobacilli stimulate the formation of a healthy intestinal
microbiota. The NDO are preferably not or only partially digested
in the intestine by the action of acids or digestive enzymes
present in the human upper digestive tract, in particular in the
small intestine and stomach, and are fermented by the human
intestinal microbiota. For example, sucrose, lactose, maltose and
the common maltodextrins are considered digestible.
[0032] Preferably the present composition comprises non-digestible
oligosaccharides with a DP in the range of 2 to 250, more
preferably 2 to 60. The non-digestible oligosaccharide is
preferably at least one, more preferably at least two, preferably
at least three selected from the group consisting of
fructo-oligosaccharides, galacto-oligosaccharides,
xylo-oligosaccharides, arabino-oligosaccharides,
arabinogalacto-oligosaccharides, gluco-oligosaccharides,
chito-oligosaccharides, glucomanno-oligosaccharides,
galactomanno-oligosaccharides, mannan-oligosaccharides, sialic acid
comprising oligosaccharides, and uronic acid oligosaccharides. The
group of fructo-oligosaccharides includes inulins, the group of
galacto-oligosaccharides includes transgalacto-oligosaccharides or
beta-galacto-oligosaccharides, the group of gluco-oligosaccharides
includes cyclodextrins, gentio- and nigero-oligosaccharides and
non-digestible polydextrose, the group of
galactomanno-oligosaccharides includes partially hydrolyzed guar
gum, and the group of uronic acid oligosaccharides includes
galacturonic acid oligosaccharides and pectin degradation
products.
[0033] More preferably the present composition comprises at least
one, more preferably at least two, most preferably three selected
from the group consisting of fructo-oligosaccharides,
beta-galacto-oligosaccharides and uronic acid oligosaccharides.
More preferably the composition comprises
beta-galacto-oligosaccharides.
[0034] In a preferred embodiment the composition comprises a
mixture of inulin and short chain fructo-oligosaccharides. In a
preferred embodiment the composition comprises a mixture of
galacto-oligosaccharides and fructo-oligosaccharides selected from
the group consisting of short chain fructo-oligosaccharides and
inulin, more preferably inulin. A mixture of at least two different
non-digestible oligosaccharides advantageously stimulates the
beneficial bacteria of the intestinal microbiota to a greater
extent. Preferably the weight ratio in a mixture of the two
different non-digestible oligosaccharides, preferably
galacto-oligosaccharides and fructo-oligosaccharide, is between 25
and 0.05, more preferably between 20 and 1.
Galacto-oligosaccharides, preferably beta-galacto-oligosaccharides,
are more capable of stimulating bifidobacteria. Preferably the
present composition comprises galacto-oligosaccharides, preferably
beta-galacto-oligosaccharides, with a degree of polymerization (DP)
of 2 to 10 and/or fructo-oligosaccharides with a DP of 2 to 60.
[0035] The galacto-oligosaccharides preferably are
beta-galacto-oligosaccharides. In a particularly preferred
embodiment the present composition comprises
beta-galacto-oligosaccharides ([galactose]n-glucose; wherein n is
an integer ranging 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, and 10),
wherein the galactose units are in majority linked together via a
beta linkage. Beta-galacto-oligosaccharides are also referred to as
trans-galacto-oligosaccharides (TOS). Beta-galacto-oligosaccharides
are for example sold under the trademark Vivinal.TM. (Borculo Domo
Ingredients, Netherlands). Another suitable source is Bi2Munno
(Classado). Preferably the TOS comprises at least 80% beta-1,4 and
beta-1,6 linkages based on total linkages, more preferably at least
90%.
[0036] Fructo-oligosaccharide is a 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.
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 commercially available, e.g. Raftiline.RTM.HP (Orafti).
Preferably the fructo-oligosaccharide has an average DP above
20.
[0037] Uronic acid oligosaccharides are preferably obtained from
pectin degradation products. Hence the present composition
preferably comprises a pectin degradation product with a DP of 2 to
100. Preferably the pectin degradation product is prepared from
apple pectin, beet pectin and/or citrus pectin. Preferably the
uronic acid oligosaccharide is a galacturonic acid oligosaccharide.
Preferably the composition comprises FL and one of the group
selected from galacto-oligosaccharide and uronic acid
oligosaccharide.
[0038] Besides FL, most preferably the composition comprises
beta-galacto-oligosaccharide, fructo-oligosaccharide and a uronic
acid oligosaccharide. It was found that such a combination acts
synergistically with fucosyllactose, in particular
2'-fucosyllactose. The weight ratio
beta-galacto-oligosaccharide:fructo-oligosaccharide:uronic acid
oligosaccharide is preferably (20 to 2):1:(1 to 20), more
preferably (20 to 2):1:(1 to 10), even more preferably (20 to
2):1:(1 to 3), even more preferably (12 to 7):1:(1 to 2). Most
preferably the weight ratio is about 9:1:1.1.
[0039] Preferably the weight ratio FL to
beta-galacto-oligosaccharide, preferably TOS, is from 5 to 0.05,
more preferably 5 to 0.1, more preferably from 2 to 0.1. Preferably
the weight ratio FL to fructo-oligosaccharide, preferably inulin,
is from 10 to 0. 05, more preferably 10 to 0.1, more preferably
from 2 to 0.5. Preferably the weight ratio FL to uronic acid
oligosaccharide, preferably derived from pectin, is from 10 to 0.
05, more preferably 10 to 0.1 more preferably from 2 to 0.5.
[0040] Preferably, the composition comprises 80 mg to 4 g
non-digestible oligosaccharides, including fucosyllactose, per 100
ml, more preferably 150 mg to 2 g, even more preferably 300 mg to 1
g non-digestible oligosaccharides per 100 ml. Based on dry weight,
the composition preferably comprises 0.25 wt % to 25 wt %
non-digestible oligosaccharides including fucosyllactose, more
preferably 0.5 wt % to 10 wt %, even more preferably 1.5 wt % to
7.5 wt %. A lower amount of non-digestible oligosaccharides will be
less effective in stimulating the beneficial bacteria in the
microbiota, whereas a too high amount will result in side-effects
of bloating and abdominal discomfort.
[0041] Nutritional Composition
[0042] Preferably the composition comprising fucosyllactose is a
nutritional composition. The composition of the present invention
is not human milk. The present composition is preferably enterally
administered, more preferably orally.
[0043] The present composition is preferably a nutritional formula,
preferably an infant formula. The present composition can be
advantageously applied as a complete nutrition for infants. The
present composition preferably comprises a lipid component, protein
component and carbohydrate component and is preferably administered
in liquid form. The present invention includes dry food, preferably
a powder, which is accompanied with instructions as to admix said
dry food mixture with a suitable liquid, preferably with water.
[0044] The present invention advantageously provides a composition
wherein the lipid component provides 5 to 50% of the total
calories, the protein component provides 5 to 50% of the total
calories, and the digestible carbohydrate component provides 15 to
85% of the total calories. The present invention advantageously
provides a composition wherein the lipid component provides 20 to
50% of the total calories, the protein component provides 5 to 30%
of the total calories, and the digestible carbohydrate component
provides 30 to 70% of the total calories. Preferably, in the
present composition the lipid component provides 35 to 50% of the
total calories, the protein component provides 7.5 to 12.5% of the
total calories, and the digestible carbohydrate component provides
40 to 55% of the total calories. For calculation of the % of total
calories for the protein component, the total of energy provided by
the proteins, peptides and amino acids needs to be taken into
account.
[0045] The present composition preferably comprises at least one
lipid selected from the group consisting of animal lipid, excluding
human lipids, and vegetable lipids. Preferably the present
composition comprises a combination of vegetable lipids and at
least one oil selected from the group consisting of fish oil,
animal oil, algae oil, fungal oil, and bacterial oil. The present
composition preferably comprises long chain poly-unsaturated fatty
acids (LC-PUFA). LC-PUFA are fatty acids or fatty acyl chains with
a length of 20 to 24 carbon atoms, preferably 20 or 22 carbon atoms
comprising two or more unsaturated bonds. More preferably the
present composition comprises eicosapentaenoic acid (EPA, n-3),
docosahexaenoic acid (DHA, n-3) and/or arachidonic acid (ARA,
n-6).
[0046] Preferably the present composition comprises 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.
[0047] The content of LC-PUFA, particularly the LC-PUFA with 20 and
22 carbon atoms, preferably does not exceed 6 wt %, more preferably
does not exceed 3 wt.% of the total fat content as it is desirable
to mimic human milk as closely as possible. The LC-PUFA may be
provided as free fatty acids, in triglyceride form, in diglyceride
form, in monoglyceride form, in phospholipid form, or as a mixture
of one of more of the above. The present composition preferably
comprises between 5 and 75 wt.% polyunsaturated fatty acids based
on total fat, preferably between 10 and 50 wt.%.
[0048] The protein used in the nutritional composition 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 thereof, free amino
acids and mixtures thereof. 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.
[0049] The present composition preferably contains digestible
carbohydrates selected from the group consisting of sucrose,
lactose, glucose, fructose, corn syrup solids, starch and
maltodextrins, more preferably lactose.
[0050] In view of the above, it is also important that the liquid
food does not have an excessive caloric density, however still
provides sufficient calories to feed the subject. Hence, the liquid
food preferably has a caloric density between 0.1 and 2.5 kcal/ml,
even more preferably a caloric density of between 0.5 and 1.5
kcal/ml, most preferably between 0.6 and 0.8 kcal/ml.
[0051] Preferably the present composition comprises nucleotides
and/or nucleosides, more preferably nucleotides. Preferably, the
composition comprises cytidine 5'-monophospate, uridine
5'-monophospate, adenosine 5'-monophospate, guanosine
5'-monophospate, and/or inosine 5'-monophospate, more preferably
cytidine 5'-monophospate, uridine 5'-monophospate, adenosine
5'-monophospate, guanosine 5'-monophospate, and inosine
5'-monophospate. Preferably the composition comprises 5 to 100,
more preferably 5 to 50 mg, most preferably 10 to 50 mg nucleotides
and/or nucleosides per 100 gram dry weight of the composition. The
presence of nucleotides and/or nucleotides advantageously
stimulates NK cell activity. The nucleotides and/or nucleosides are
deemed to act synergistically with the fucosyllactose of the
present composition.
[0052] Application
[0053] In one embodiment the present composition is used for
stimulating natural killer cell activity and/or natural killer cell
proliferation. In one embodiment the present composition is used
for treating and/or preventing viral infections. In one embodiment
the composition for stimulating natural killer cell activity and/or
natural killer cell proliferation, and/or for treating and/or
preventing viral infections is for administering to HIV patients,
elderly and/or oncology patients.
[0054] NK cells are a type of cytotoxic lymphocytes that constitute
a major component of the innate immune system. NK cells play a
major role in defense against intracellular infections. NK-cells
are defined as large granular lymphocytes that do not express
T-cell antigen receptors (TCR) or Pan T marker CD3 or surface
immunoglobulins (Ig) B cell receptor but that usually express the
surface markers CD16 (Fc.gamma.RIII) and CD56 in humans. They were
named "natural killers" because of the initial notion that they do
not require activation in order to kill cells that are missing
"self' markers of major histocompatibility complex (MHC) class I.
NK cells have two major types of effector function; cell killing
and the secretion of cytokines. Increasing NK cell activity (by
increasing the number of NK cells and/or by increasing the specific
activity of an NK cell), results in an increased resistance against
viral infections. The use of a nutritional composition comprising
fucosyllactose is therefore preferably for preventing and/or
treating 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 a nutritional composition comprising
fucosyllactose is therefore preferably for preventing and/or
treating viral infections, more preferably the viral infections
common cold, flu, measles, chicken pox, viral diarrhoea, viral
gastro-enteritis, HIV infection and/or viral respiratory tract
infections. In a preferred embodiment the present invention is used
for HIV patients. In one embodiment, the present invention concerns
providing nutrition to a HIV patient. The present composition is
advantageous for HIV patients since HIV patients have a decreased
natural killer cell activity.
[0055] The use of a nutritional composition comprising
fucosyllactose is therefore especially beneficial for infant
formula. In one embodiment, the present invention concerns
providing nutrition to an infant. Formula fed infants have an
underdeveloped immune system compared with adults and are more
prone to viral infections than human milk fed infants. Preferably
the infant is from 0 to 36 months of age, more preferably of 0 to
18 months, even more preferably of 0 to 12 months, most preferably
of 0 to 6 months of age. The younger the infant is, the less
developed the immune system.
[0056] The composition comprising fucosyllactose even more
advantageously is used in preterm infants and/or very low or low
birth weight infants, since these infants are even more vulnerable
and/or prone to viral infections.
[0057] The composition comprising fucosyllactose even more
advantageously is used in infants delivered via Caesarean section.
Caesarean section born infants are born in a hospital in an
environment having more pathogens against which the antibodies,
conferred by the mother to the infant, are not effective against.
Caesarean section born infants have a delayed and less optimal
colonization of the large intestinal tract and therefore are also
more prone to intestinal infections.
[0058] The composition comprising fucosyllactose is advantageously
used for nutrition for elderly. In one embodiment, the present
invention concerns providing nutrition to an elderly person. An
elderly person is a person having an age of 55 years or more, in
particular of the age of 65 or more. Elderly have a demonstrated
lower activity of natural killer cell activity than healthy young
adult individuals. Elderly are especially vulnerable to viral
infection complications. In a preferred embodiment the present
invention is used for treatment and/or prevention of
immunosenescence in elderly. Elderly are more prone to the
development of tumours. NK cell activity suppresses tumour cell
proliferation. In a preferred embodiment the present invention is
used for nutrition in cancer patients. Oncology patients have a
lower natural killer cell activity than healthy young adult
individuals.
EXAMPLES
Example 1
[0059] Materials and Methods
[0060] 6-8 Weeks old female C57BL/6 mice (Charles River) received
semi-purified AIN-93G-based diets (Research Diet Service, Wijk bij
Duurstede, the Netherlands), comprising 1) 2 wt %
beta-galacto-oligosaccharide (GOS; source Vivinal GOS, Borculo
Domo), fructo-oligosaccharide (FOS; source RaftilineHP, Orafti) and
galacturonic acid oligosaccharide (Source AOS) in a 9:1:1.1 ratio.
AOS are produced from pectin (Sudzucker AG, Mannheim, Germany),
with a DP of 1-20. It consists of approximately 75% galacturonic
acid oligomers, based on total weight;
[0061] 2) 1 wt % lactoneotetraose (LNnT),
[0062] 3) 1 wt % 3'-sialyl lactose (3'-SL), or
[0063] 4) 1 wt % 2'-fucosyllactose (2'-FL).
[0064] All groups were compared to the unsupplemented control diet.
Dietary supplementation started 14 days before the first
vaccination and lasted until the end of the experiment, 31 days
after the first vaccination.
[0065] Vaccination experiments were performed using Influvac
(Solvay Pharmaceuticals, Weesp, the Netherlands) from season
2005/2006. The mice received a primary vaccination and a booster
vaccination, consisting of a subcutaneous (sc) injection of a 1:1
mix of vaccine and adjuvant in a total volume of 100 .mu.l. The
booster vaccination was given at 21 days after the primary
vaccination. The experiments ended 10 days after booster
vaccination. Blood samples were taken at the end of the experiment.
Negative control groups that were included received injections with
a 1:1 mix of PBS and adjuvant in a total volume of 100 .mu.l. To
determine the percentage of NK cells, cells were labelled with
FITC-labelled anti-mouse CD3 mAb in combination with PE-labelled
anti-mouse NK1.1 mAb. NK cell cytotoxicity in spleen cell
suspensions was assayed using standard .sup.51Cr release assays.
Briefly, NK cell cytotoxicity was tested using YAC-1 target cells.
The percentage of specific .sup.51Cr release was calculated as the
percentage of specific lysis=(experimental release-spontaneous
release)/(total detergent release-spontaneous release).times.100.
The spontaneous release values were always <15% of total
lysis.
[0066] The percentage of regulatory T cells (Treg) was determined
by flow cytometry (FACSCalibur) using allophycocyanin
(APC-)labelled anti-mouse CD3 mAb, Pe-Cy5-labeled anti-mouse CD4
mAb and phycoerythrin (PE)-labelled anti-mouse CD25 mAb in
combination with intracellular staining of
fluorescein-isotiocyanate (FITC)-labelled Foxp3 mAb, according to
the instructions offered by the manufactures (Biosciences, San
Diego, Calif.).
[0067] Statistical analysis was performed using GraphPadPrism
software. Statistical differences between test and control groups
were analysed by ANOVA and post hoc Dunnett's test if multiple
groups were compared to a single (control) group. P-values <0.05
were considered significant in all experiments.
[0068] Results
[0069] The immunomodulatory effect of three chemically synthesised
human oligosaccharides was compared with GOS/FOS/AOS.
Supplementation with GOS/FOS/AOS, LNnT, 2'-FL or 3'-SL resulted in
a significant increase of the DTH response, a TH1-dependent
parameter, compared with control-fed animals.
[0070] Interestingly, the percentage of NK cells in the spleen was
significantly increased in mice supplemented with human
oligosaccharides compared to control and GOS/FOS/AOS-supplemented
mice, see table 1. This effect was highest with 2'-FL. To examine
whether the increase in the percentage of NK cells in human
oligosaccharides-supplemented groups also correlated with
functional activity, NK cell activity was measured in mouse
splenocytes. A significant increased NK cell activity was detected
in the splenocytes of mice that were supplemented with 2'-FL
compared to controls, see Table 1.
TABLE-US-00001 TABLE 1 Effect of dietary NDO similar to human milk
NDO on NK cells and NK cell activity. NK cell activity, Dietary %
NK 1.1 + % lysis At E:T intervention cells (SE) ratio 1:50 Sham
control 2.5 (0.2) 1.57 Control 2.2 (0.1) 1.13 GOS/FOS/AOS 2.5 (0.1)
1.95 LNnT 2.8 (0.1)* 2.14 3'-SL 3.0 (0.2)* 1.93 2'-FL 3.7 (0.1)**
2.48* *indicates .rho. < 0.05 compared to control group
**indicates .rho. < 0.01 compared to control group
[0071] Finally, the amount of regulatory T cells (Treg) was
decreased to the highest extent with the diet with FL as can be
seen in Table 2.
TABLE-US-00002 TABLE 2 Effect of dietary NDO similar to human milk
NDO on percentage of regulatory T cells Treg. Dietary Treg
percentage intervention (s.e.m.) Sham control 3.6 (0.4) Control 3.9
(0.2) GOS/FOS/AOS 4.1 (0.3) LNnT 3.2 (0.2) 3'-SL 2.8 (0.2)* 2'-FL
2.8 (0.2)* *indicates .rho. < 0.05 compared to control group
[0072] The decrease in regulatory T cells is indicative for a
decreased inhibition of immune response and hence enables an
increased vaccination response to the viral antigens. A temporarily
decrease of regulatory T cells can be especially beneficial when
vaccination is to occur.
[0073] Overall, these results support that oral supplementation
with 2'-FL stimulates growth and/or activity of NK cells. These
results are indicative for an effect of dietary 2'-FL for enhancing
vaccination response, in particular vaccination with viral
antigens. These results are indicative for an effect of dietary
2'-FL for treating and/or preventing viral infections.
Example 2
[0074] Infant formula for stimulating NK cell activity comprising
per 100 ml (13.9 dry weight):
[0075] 1.4 g protein (whey and casein)
[0076] 7.3 g digestible carbohydrates (including lactose)
[0077] 3.6 g fat (vegetable fat, fish oil)
[0078] 0.8 g non-digestible oligosaccharides of which 80 mg
2'-fucosyllactose and 640 mg beta-galacto-oligosaccharides, and 80
mg fructo-oligosaccharides
[0079] Further are included: choline, myo-inositol, taurine,
minerals, trace elements, and vitamins as known in the art.
Example 3
[0080] A preferred composition that can be used for the stimulation
of natural killer cell activity in HIV patients may comprise per
100 g dry weight.
TABLE-US-00003 Dietary fibre 5-50 g Fructo-oligosaccharide 5% of
total dietary fibre Galacto-oligosaccharide 40% of total dietary
fibre Pectin hydrolysate 50% of total dietary fibre 2'-FL 5% of
total dietary fibre N-acetyl cysteine 0.5-5 g Carbohydrate (not
dietary fibre) 2-20 g Fat 4-20 g
* * * * *