U.S. patent application number 12/282302 was filed with the patent office on 2009-12-10 for use of non-digestable sacharides for giving an infant the best start after birth.
This patent application is currently assigned to N.V. Nutricia. Invention is credited to Martine Sandra Alles, Christopher Beermann, Gunther Boehm, Jan Knol, Bernd Stahl.
Application Number | 20090305996 12/282302 |
Document ID | / |
Family ID | 38349460 |
Filed Date | 2009-12-10 |
United States Patent
Application |
20090305996 |
Kind Code |
A1 |
Beermann; Christopher ; et
al. |
December 10, 2009 |
USE OF NON-DIGESTABLE SACHARIDES FOR GIVING AN INFANT THE BEST
START AFTER BIRTH
Abstract
A food or supplement for pregnant women comprising water
soluble, non-digestible saccharides is described. The composition
is used to improve the flora and/or immune system of the pregnant
women, to improve the immune system of the infant and to improve
the intestinal flora of the infant after birth.
Inventors: |
Beermann; Christopher;
(Neu-Anspach, DE) ; Knol; Jan; (Wageningen,
NL) ; Alles; Martine Sandra; (Apeldoorn, NL) ;
Stahl; Bernd; (Rosbach-Rodheim, DE) ; Boehm;
Gunther; (Echzell, DE) |
Correspondence
Address: |
FOLEY AND LARDNER LLP;SUITE 500
3000 K STREET NW
WASHINGTON
DC
20007
US
|
Assignee: |
N.V. Nutricia
|
Family ID: |
38349460 |
Appl. No.: |
12/282302 |
Filed: |
March 9, 2007 |
PCT Filed: |
March 9, 2007 |
PCT NO: |
PCT/NL2007/050094 |
371 Date: |
February 9, 2009 |
Current U.S.
Class: |
514/23 |
Current CPC
Class: |
A61P 11/06 20180101;
A61P 15/00 20180101; A61P 37/08 20180101; A61P 1/00 20180101; A61P
37/00 20180101; A61K 31/702 20130101; A61P 11/00 20180101; A61P
37/02 20180101; A23L 33/115 20160801; A61P 1/10 20180101; A23V
2002/00 20130101; A23L 33/30 20160801; A61P 21/02 20180101; A61P
27/16 20180101; A61P 37/04 20180101; A23L 33/21 20160801; A61P 1/14
20180101; A23V 2002/00 20130101; A23V 2200/324 20130101; A23V
2250/28 20130101; A23V 2250/1882 20130101; A61K 31/702 20130101;
A61K 2300/00 20130101 |
Class at
Publication: |
514/23 |
International
Class: |
A61K 31/70 20060101
A61K031/70; A61K 31/702 20060101 A61K031/702; A61P 37/00 20060101
A61P037/00 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 10, 2006 |
EP |
06110973.2 |
Apr 20, 2006 |
EP |
06112822.9 |
Claims
1-11. (canceled)
12. A method of improving the intestinal flora development of an
infant after birth; strengthening the immune system of an infant
before or after birth; and/or preventing immune system related
disorders of an infant before birth, the method comprising
administering to a woman pregnant with the infant, a composition
comprising a first water soluble, non-digestible saccharide.
13. The method according to claim 12, wherein the composition is
administered to a woman pregnant of an infant for prevention of at
least one disorder selected from the group consisting of allergy,
eczema, asthma, rhinitis, hayfever, rhinoconjunctivitis, wheezing,
intestinal infections, vaginal infections, respiratory infections,
diarrhoea, constipation, cramps and intestinal inflammation in the
infant after birth.
14. The method according to claim 12, wherein the saccharide is
administered in a dose of 0.5 to 50 g/day, during at least 2 weeks
before due delivery.
15. The method according to claim 12, wherein bacterial
colonisation of the intestine of the infant is stimulated; and/or
wherein the percentage of bifidobacteria and/or lactobacilli based
on total bacteria in the colon and/or faeces of the infant after
birth is enhanced.
16. The method according to claim 12, wherein the saccharide
comprises at least 50% galactose monosaccharide units based on the
total number of monosaccharide units present in the saccharide.
17. The method according to claim 12, wherein the saccharide is
selected from the group consisting of transgalactooligosaccharides,
galactooligosaccharides, lacto-N-tetraose (LNT),
lacto-N-neotetraose (neoLNT), fucosyl-lactose, fucosylated LNT and
fucosylated neo-LNT, and alpha-galacto-oligosaccharide, preferably
transgalactooligosaccharide.
18. The method according to claim 12, wherein the composition
further comprises a second water soluble, non-digestible
saccharide, wherein the first and the second saccharides have i)
different glycosidic linkages, ii) different degree of
polymerisation, and/or iii) different monosaccharide
composition.
19. The method according to claim 17, wherein the first saccharide
comprises at least 50% of galactose units based on total number of
monosaccharide units present in the first saccharide and wherein
the second saccharide is selected from the group consisting of
fructopolysaccharides, fructooligosaccharides, and inulin.
20. The method according to claim 19, wherein the second saccharide
is inulin.
21. A method of strengthening the immune system of a pregnant
woman; improving the intestinal flora of a pregnant woman; and/or
improving the vaginal flora of a pregnant woman, the method
comprising administering to the pregnant woman a composition
comprising a water soluble, non-digestible saccharide, wherein the
saccharide comprises at least 50% galactose units based on the
total number of monosaccharide units present in the saccharide.
22. The method according to claim 21, wherein the composition
further comprises a saccharide selected from the group consisting
of fructopolysaccharide, fructo-oligosaccharide, and inulin.
23. The method according to claim 12, wherein the saccharide is
inulin.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to a composition for
stimulating health of an infant by administering the composition to
a pregnant woman.
BACKGROUND OF THE INVENTION
[0002] For the mother it is particularly important that the baby
develops well during pregnancy and gets an optimal start when it is
born. Hence, pregnant women often ingest nutritional compositions
which aim to improve growth and development of the unborn
child.
[0003] Long chain poly-unsaturated fatty acids (LC-PUFA) have been
described to stimulate the brain development of the unborn child.
For that reason, pregnant (and lactating) women ingest nutritional
compositions with LC-PUFA. EP705539 describes such a
composition.
[0004] EP1296694 relates to prophylaxis of allergies, and relates
specifically to primary prevention of atopic diseases by
administering probiotic bacteria, beneficial microbes present in a
healthy gut flora, pre- and postnatal to children at high risk of
atopic diseases.
SUMMARY OF THE INVENTION
[0005] The inventors have found that administration of a
water-soluble, non-digestible saccharide to pregnant women improves
the immune system of the unborn infant. It was found that the
unborn child has an enhanced expression of the receptors CD14, and
TLR 2 as present in the membrane of enterocytes and monocytes.
[0006] Additionally, it was found that the development of the
intestinal flora of the infant delivered by a mother who has
ingested the present saccharide is improved compared to the
development of the intestinal flora of an infant delivered by a
mother that did not ingest the present saccharide.
[0007] The inventors have found that the intestinal flora
development of the infant is improved as a result of an improved
intestinal and/or vaginal flora of the mother ingesting the present
saccharide. After ingestion of the present saccharides, the growth
of bifidobacteria and lactobacilli in the gastrointestinal tract of
the mother is stimulated. Subsequently, by cross colonisation, the
vaginal flora of the mother is also beneficially changed towards a
high content and diversity of the bifidobacteria and lactobacilli.
During birth, the neonatal gut is first inoculated by maternal
vaginal and intestinal microflora. Hence, an advantageous vaginal
and intestinal flora of the mother results in an improved flora of
the infant in the first period of life.
[0008] As a result of the stimulated immune system and optimal
intestinal flora dominated by the genera Bifidobacterium and
Lactobacillus the infant has reduced chances to develop immune
related disorders, such as atopic diseases, but also
gastro-intestinal disorders, such as infections, intestinal
inflammation, diarrhoea or constipation.
[0009] In a further aspect it was surprisingly found that
administration of this non-digestible saccharide beneficially
affected the immune system of the pregnant women and/or of the
infants. It was found that the child immune system before delivery
is primed towards a faster Th1 response after birth. As a result
the chances to develop immune related disorders, such as atopic
diseases, is even further reduced.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
[0010] The present invention provides the use of a water soluble,
non-digestible saccharide for the manufacture of a composition for
[0011] i) improving the intestinal flora development of an infant
after birth; [0012] ii) strengthening the immune system of an
infant before birth; and/or; [0013] iii) strengthening the immune
system and/or prevention of immune system related disorders of the
infant after birth; wherein the composition is administered to a
woman pregnant of said infant.
[0014] The present invention further provides the use of water
soluble, non-digestible saccharide for the manufacture of a
composition for [0015] i) strengthening the immune system of a
pregnant woman; [0016] ii) improving the intestinal flora of a
pregnant woman; and/or [0017] iii) improving the vaginal flora of a
pregnant woman; wherein the composition is administered to the
pregnant woman.
Water Soluble, Non-Digestible Saccharide
[0018] The composition used in the present method comprises water
soluble, non-digestible saccharides. The term "non-digestible" as
used in the present invention refers to saccharides which are not
digested in the intestine by the action of acids or digestive
enzymes present in the human upper digestive tract (small intestine
and stomach), but which are fermented by the human intestinal
flora. Mono- and disaccharides such as glucose, galactose,
fructose, sucrose and lactose are digestible. The term
"fermentable" as used herein refers to the capability to undergo
conversion by micro-organisms in the lower part of the
gastro-intestinal tract (e.g. colon) to smaller molecules, in
particular short chain fatty acids and lactate. The present
saccharide is water-soluble, which can be determined with the
method described by L. Prosky et al, J. Assoc. Anal. Chem. 71:
1017-1023, 1988.
[0019] The present saccharides preferably have a degree of
polymerisation (DP) of at least 2 and preferably below 100
monosaccharide units, preferably below 60, even more preferably
below 40, most preferably below 10. Preferably, the present water
soluble, non-digestible saccharide is a galactose comprising
saccharide. A saccharide with a relatively low degree of
polymerisation has an improved fermentability by lactobacilli
and/or bifidobacteria and further does not have the technological
disadvantage to increase the viscosity.
Water Soluble, Non-Digestible Galactose Comprising Saccharide
[0020] The present saccharide is preferably a water soluble,
non-digestible galactose containing saccharide (hereinafter
referred to as "GAL-oligo"), which preferably comprise at least 50%
galactose units based on the total number of monosaccharide units
of the saccharide. GAL-oligo has an improved bifidogenic effect
compared to other water soluble, non-digestible saccharides. Human
breast milk also comprises a high concentration of GAL-oligo and
infants fed human breast milk have an intestinal flora more rich in
bifidobacteria than infants fed a standard infant milk formula.
Preferably the present GAL-oligo contains at least 60% galactose
units based on the total number of monosaccharide units present in
the saccharide, more preferably, at least 65%.
[0021] The present GAL-oligo preferably comprises at least two
terminal saccharide units, wherein at least one terminal saccharide
unit is selected from the group consisting of glucose and
galactose; and at least one terminal saccharide unit is selected
from the group consisting of galactose and fucose. Preferably at
least 75% of the saccharides of the GAL-oligo are .beta.-linked,
preferably 100%.
[0022] The term "terminal saccharide" refers to a saccharide which
is bound to one other saccharide unit (e.g. galactose, glucose,
fructose or fucose). The present GAL-oligo preferably contains not
more than 4 terminal saccharides, preferably not more than 2. In a
preferred embodiment, the GAL-oligo comprises at least one terminal
galactose and one selected from at least terminal glucose and one
terminal fucose. Even more preferably, the present GAL-oligo
comprises at least one terminal galactose and at least one terminal
glucose. Preferably the present Gal-oligo contains 2 terminal
saccharide units and has a degree of polymerisation (DP) of 2 to
60.
[0023] Preferably the GAL-oligo is selected from the group
consisting of transgalactooligosaccharides,
galactooligosaccharides, lacto-N-tetraose (LNT),
lacto-N-neotetraose (neo-LNT), fucosyl-lactose, fucosylated LNT and
fucosylated neo-LNT. Preferably the GAL-oligo is sialylated.
Preferably the GAL-oligo is selected from the group consisting of
sialyllactose, sialylfucosyllactose, sialyllactosamine and
sialyl-LNT and sialyl-neo-LNT, more preferably sialyllactose. In a
particularly preferred embodiment the present method comprises the
administration of transgalactooligosaccharides
([galactose].sub.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). Preferably the saccharides of
the transgalactooligosaccharides are .beta.-linked.
Transgalactooligosaccharides (TOS) are for example present in a
composition sold under the trademark Vivinal.TM. (Borculo Domo
Ingredients, Netherlands). Other suitable sources of TOS are
Cup-oligo and Bi.sup.2muno.
Different Water Soluble, Non-Digestible Saccharides
[0024] In order to achieve an even better effect regarding
intestinal flora and/or (improvement of) the immune system of the
infant or of the intestinal flora, vaginal flora and/or
(improvement of) the immune system of the pregnant woman, the
present composition preferably comprises two different water
soluble, non-digestible saccharides, differing from each other in
structure. Each water soluble, non-digestible saccharide is
fermented by different species of micro-organisms in the intestinal
and/or vaginal flora and/or on a different location in the
intestinal tract, resulting in an improved flora and greater
diversity of bacteria.
[0025] The present invention provides a composition which
preferably comprises water soluble, non-digestible saccharide A and
water soluble, non-digestible saccharide B. Saccharide A and
saccharide B have different glycosidic linkages, a different degree
of polymerisation and/or a different monosaccharide composition.
Preferably, saccharide A is a GAL-oligo.
[0026] According to a preferred embodiment of the present
invention, the percentage of at least one monosaccharide unit
selected from the group consisting of glucose, fructose and
galactose in saccharide A is at least 40% higher than the
percentage of the same monosaccharide unit in saccharide B,
preferably at least 50%, more preferably at least 75%, even more
preferably at least 90%. An increased diversity of monosaccharide
units stimulates a wider population of intestinal (beneficial)
bacteria, resulting in an improved flora. The percentage of a
monosaccharide unit in the saccharide can be simply calculated by
dividing the number of the respective monosaccharide units (e.g.
glucose) present in the saccharide by the total number of the
monosaccharide units present in that saccharide.
[0027] Preferably water soluble, non-digestible saccharide A and B
have a degree of polymerisation (DP) between 2 and 200. Preferably
at least 80 wt. %, more preferably at least 95 wt. %, most
preferably at least 98 wt. % of the cumulative weight of water
soluble, non-digestible saccharide A and B has a degree of
polymerisation (DP) below 100, more preferably below 60, most
preferably below 40. The lower DP advantageously reduces viscosity
and increases fermentability of the non-digestible saccharides.
Preferably, at least 50 wt. %, more preferably at least 75 wt. % of
the cumulative weight of water soluble, non-digestible saccharides
A and B has a DP of 2 to 8. By using a mixture with a high weight
percentage of saccharides with a low DP the fementability and
stimulatory effect on the growth of the lactic acid bacteria and
bifidobacteria will be increased.
[0028] According to a preferred embodiment of the present
invention, the DP of water soluble, non-digestible saccharide A is
at least 5 monosaccharide units lower than the DP of water soluble,
non-digestible saccharide B, preferably at least 10, even more
preferably at least 15. Including a saccharide with an increased
degree of polymerisation reduces the osmotic load, and results in a
prolonged fermentation along the colon, thereby improving
stimulation of beneficial bacteria also in more distal parts of the
colon. Preferably, water soluble, non-digestible saccharide A has a
DP of 2-10, more preferably 2-8. Preferably the water soluble,
non-digestible saccharide B has DP of 11-100. The water soluble,
non-digestible saccharides A and B with a different DP may have the
same or slightly different monosaccharide composition, preferably
different monosaccharide compositions.
[0029] In a preferred embodiment of the present invention the
percentage of at least one glycosidic linkage of water soluble,
non-digestible saccharide A based on total glycosidic linkages of
present in saccharide A is at least 40% higher than the percentage
of the same glycosidic linkage in water soluble, non-digestible
saccharide B, preferably at least 50%, even more preferably at
least 75%. The term "glycosidic linkage" as used in the present
invention refers to a C--O--C bond formed between the rings of two
cyclic monosaccharide units by the elimination of water. An
increased diversity in glycosidic linkages stimulates a wider range
of beneficial bacteria. Glycosidic linkages differ in that they
covalently bind carbon atoms in the monosaccharide units at
differently numbered positions, and/or that they form a or P bonds.
Examples of different glycosidic linkages occurring in water
soluble, non-digestible saccharides are .beta.(1,3), .alpha.(1,4),
.beta.(2,1), .alpha.(1,2), and .beta.(1,4) linkages. Preferably the
glycosidic linkages in water soluble, non-digestible saccharide A
comprises at least 40% .beta.(1,4) glycosidic linkages, more
preferably at least 75%. The glycosidic linkages in water soluble,
non-digestible saccharide B preferably comprise at least 40%
.beta.(2,1) glycosidic linkages, more preferably at least 75%.
[0030] In a preferred embodiment the present composition contain a
combination of Gal-oligo and a water soluble, non-digestible
saccharide selected from the group consisting of
fructopolysaccharides (such as inulin), more preferably a
combination of GAL-oligo which have an average DP between 2 and 10
and a water soluble, non-digestible saccharide selected from the
group consisting of fructopolysaccharides and
fructooligosaccharides, most preferably a combination of
transgalactooligosaccharides and inulin.
[0031] Preferably saccharide A and saccharide B are in a weight
ratio of 3/97 to 97/3, more preferably 5/95 to 95/5. Having both
saccharide A and B present in a sufficient relative proportion will
have a better effect on flora. Most preferably the weight ratio of
saccharide A to B is 2/3 to 95/5. A slight excess of saccharide A,
being preferably a GAL-oligo and/or an oligosaccharide, is even
further preferred as this will have a better effect on the
flora.
Long Chain Polyunsaturated Fatty Acids (LC-PUFA)
[0032] Preferably, the present composition additionally comprises
long chain polyunsaturated fatty acids and/or long chain
polyunsaturated fatty acyl chains (LC-PUFA). The term LC-PUFA in
the present invention relates to a fatty acid or fatty acyl chain
with a length of 20 carbons or more and at least two unsaturated
bonds. Preferably, the composition comprises a LC-PUFA selected
from the group consisting of eicosapentaenoic acids and/or acyl
chain (EPA), docosahexaenoic acid and/or acyl chain (DHA) and
arachidonic acid and/or acyl chain (AA). In one embodiment the
composition comprises DHA. In another embodiment the composition
comprises AA. More preferably the composition comprises DHA and AA.
LC-PUFA advantageously strengthen the immune system of the infant
to be born. Hence, administration of a composition comprising both
a water soluble, non-digestible oligosaccharide and LC-PUFA to a
pregnant women will have an improved effect on the immune system of
the infant compared to the water soluble, non-digestible saccharide
alone.
[0033] The weight ratio EPA/DHA is preferably 1 or lower, more
preferably below 0.5. The LC-PUFA may be provided as free fatty
acids, in triglyceride form, in phospholipid form, or as a mixture
of one of more of the above. The present composition preferably
comprises at least one of AA and DHA in phospholipid form.
Preferably the weight ratio DHA to AA is between 0.1 and 20, more
preferably between 0.5 and 5.
[0034] Preferably the composition comprises 50 to 3000 mg LC-PUFA
per daily dose, more preferably 100 to 1500 mg, most preferably 200
to 500 mg.
Other Components
[0035] In a preferred embodiment the composition additionally
comprises probiotics. Probiotics in the present invention refer to
micro-organisms, which upon administration exert a beneficial
effect on the host. Probiotics have beneficial effects on
intestinal flora, vaginal flora and/or the immune system of
pregnant women and their infant to be born, hence the combination
of a water soluble, non-digestible saccharide and probiotics will
have a superior effect on intestinal flora and/or immune system of
both. Preferably, the probiotics are selected from the group
consisting of Lactobacillus and Bifidobacterium. More preferably,
the probiotic is selected from the group consisting of the
Lactobacillus accidophilus group, L. rhamnosus, L. casei, L.
paracasei, L. plantarum, L. reuteri, L. fermentum, Bifidobacterium
infantis, B. animalis subsp. lactis, B. breve, B. longum, and B.
bifidum. Even more preferably, the probiotics are L. paracasei
and/or B. breve, since administration of water soluble,
non-digestible saccharide stimulates most species of Lactobacillus
and Bifidobacterium population to an equal extent, but stimulates
B. breve and L. paracasei to a lesser extent. Since bifidobacteria
are more dominant in the infant intestinal flora than lactobacilli,
B. breve is preferred most. Preferably, probiotics are present in a
daily dose of 1.times.10.sup.6 colony forming units (cfu) to
1.times.10.sup.13 cfu, more preferably 1.times.10.sup.7 to
1.times.10.sup.1, most preferably 1.times.10.sup.8 to
1.times.10.sup.10 of each different probiotic.
[0036] The composition comprises additionally vitamins and minerals
beneficial for pregnant women. Often, supplements comprising
vitamins and minerals are ingested by pregnant women for their or
the foetus' benefit. Inclusion of these vitamins and minerals in
the present composition conveniently reduces the amount of
compositions to be taken by pregnant women. Preferably, the
composition comprises at least one component, more preferably at
least three components, most preferably at least six components
selected from the group consisting of folic acid, vitamin B1,
vitamin B2, vitamin B6, vitamin A, vitamin D, iron, zinc, and
iodine. Preferably, these components are present in a daily dose of
25 to 100% of the recommended daily allowance (RDA), more
preferably between 45 to 100%. Preferably vitamin A is supplied as
.beta.-carotene.
The Composition
[0037] The composition comprising the water soluble, non-digestible
saccharide may be administered to the pregnant women in the form of
a bar, a capsule, a tablet, a liquid, or a powder.
[0038] Preferably, the composition is a milk-based liquid,
comprising fats, proteins and digestible carbohydrates. Preferably
the fat content of the milk-based liquid is less than 2 g/l in
order to keep the amount of calories to be consumed low. Preferably
this milk-based liquid is packed into a bottle or tetrapack with a
volume of 50 to 1000 ml, more preferably 60 to 500, most preferably
75 to 125 ml. Preferably, the composition is a bar, i.e. a solid,
chewable composition with a water activity below 0.8, preferably
below 0.65. Preferably, the composition is a powder, packed in
sachet comprising 1 to 10 g, more preferably 1.5 to 7 g, most
preferably 2 to 5 g.
Dose
[0039] In the present method, preferably the present saccharide is
administered to the pregnant women in an amount between 0.5 and 50
g per day, preferably between 3 and 25 g per day, most preferably
between 6 and 12 g per day. Preferably, this daily dose is
administered in one portion per day. Preferably, this daily dose is
divided over 2 or 3 or 4 portions, which are consumed 2, 3 or 4
times per day, respectively.
[0040] Preferably, the composition is administered to the pregnant
women at least 2 weeks before due delivery, more preferably at
least 6 weeks, most preferably at least 12 weeks.
Applications
[0041] Depending on the jurisdiction it is to be understood that in
the part hereinbelow where a method of a certain treatment is
mentioned, said method comprising administering an effective amount
of a composition as described hereinabove or administering an
effective amount of at least a water soluble, non-digestible
saccharide to a pregnant woman, this also relates to the use of a
at least a water soluble, non-digestible saccharide for the
preparation of a composition for said purpose. Both manners of
putting the present invention to words is also covered by the
wording such as the present composition is used for a certain
purpose or the administration of the present composition is used
for a certain purpose.
[0042] A method for improving the intestinal flora and/or the
immune system of an infant is provided, said method comprising
administering the composition of the present invention, i.e. a
composition comprising water soluble, non digestible saccharides,
to a woman pregnant of said infant.
[0043] In one embodiment the immune system of the infant is
strengthened. Strengthening the immune system in the present
invention relates to improvement, stimulation and/or enhancement of
the immune system. Preferably the improvement, stimulation and/or
enhancement is with respect to a control group, which in the
context of this invention means with respect to a pregnant woman
that did not ingest water soluble, non-digestible saccharide. The
inventors observed that administration of a water soluble,
non-digestible saccharide directed the neonatal immune system
towards a Th1 response and/or improved the Th1/Th2 balance. It was
found that before delivery the immune system of the unborn child is
primed by enhancing the receptors CD14 and TLR 2.
[0044] In one embodiment, administration of water soluble,
non-digestible saccharide to pregnant women is used for preventing
the onset of atopic diseases in the infant. In one embodiment the
composition is administered to pregnant women in order to prevent
the incidence and/or reduce the severity of allergy (i.e. food
allergy), eczema (i.e. atopic dermatitis), asthma, rhinitis,
hayfever, rhinoconjunctivitis, and/or wheezing in the infant, most
preferably allergy and/or atopic dermatitis. Eczema, i.e. atopic
dermatitis, is characterized by a dry and/or red and/or an itchy
skin.
[0045] In one embodiment administration of the water-soluble,
non-digestible saccharide to the pregnant women, is used to improve
the intestinal flora of the infant. Administration of the present
composition is in an embodiment used for enhancing the percentage
bifidobacteria and/or lactobacilli in the colon and/or faeces based
on total bacteria present in the flora and/or faeces of the infant
after birth. A flora rich in bifidobacteria and/or lactobacilli
strengthens the immune system of the infant and improves the
gastro-intestinal health. An improved gastro-intestinal health
relates to a reduced incidence of gastro-intestinal infections, a
shorter duration of intestinal infections, a reduced incidence and
severity of gastro-intestinal inflammation, a reduced incidence
and/or severity of diarrhoea, reduced constipation, or reduced
cramps compared to the gastro-intestinal health of an infant of
which the mother during pregnancy did not ingest the present
composition comprising water-soluble, non-digestible saccharide.
Strengthening or strengthened and improving or improved in the
context of this invention is relative to an infant of which the
mother during pregnancy did not ingest the present composition
comprising water-soluble, non-digestible saccharide or relative to
a pregnant woman that did not ingest the present composition
comprising water-soluble, non-digestible saccharide. A strengthened
immune system will have a preventive effect on atopic diseases
and/or respiratory infections. In one embodiment administration of
the present composition to a pregnant woman is used to prevent
and/or reduce the severity of disorders selected from the group
consisting of allergy (i.e. food allergy), eczema (i.e atopic
dermatitis), asthma, rhinitis, hayfever, rhinoconjunctivitis,
wheezing, intestinal infections, vaginal infections, respiratory
infections, diarrhoea, constipation, cramps and intestinal
inflammation in the infant after birth. In one embodiment
administration of the present composition to a pregnant woman is
used to prevent disorders selected from the group consisting of
intestinal infections, respiratory infections, diarrhoea,
constipation, cramps and/or intestinal inflammation of the infant
after birth.
[0046] Packages comprising the present composition according to the
invention with a text stating that upon consumption of the
composition a pregnant woman will give her child the best start
after birth, for example regarding intestinal colonisation,
intestinal flora, flora composition and/or immune system, e.g.
immunological defense or strengthened or improved or enhanced or
stimulated immunity are also encompassed.
[0047] Administration of the composition of the invention is in one
embodiment used to improve the immune system of the pregnant
women.
[0048] Administration of the composition of the present invention,
comprising water soluble, non-digestible saccharides, preferably
comprising two different water soluble, non-digestible saccharides,
is in one embodiment used to strengthen the immune system, the
intestinal flora and/or the vaginal flora of a pregnant woman. In
an advantageous embodiment administration of the present
composition, preferably comprising two different water soluble,
non-digestible saccharides, is used to enhance the percentage of
lactobacilli and/or bifidobacteria based on total bacteria present
in the intestine and/or vagina of the pregnant women. This is of
importance since a disturbed vaginal flora is frequently found
during pregnancy and the overgrowth of pathogens and breakdown of
the vaginal microbial microenvironment can be prevented by a stable
colonization with lactobacilli.
EXAMPLES
Example 1
Clinical Trial
[0049] CA prospective, double-blind, randomized, placebo controlled
study with a parallel group design was performed with a total of 33
pregnant healthy volunteers. They received either the experimental
supplement or a placebo from the 25th week of gestation until
delivery. The experimental supplement was based on non-digestible
oligosaccharides, and comprised 3 g of GOS (from Vivinal GOS,
Borculo Domo, The Netherlands) plus inulin (RaftilinHP, Orafti,
Belgium), in a 9:1 wt/wt ratio and 3 g of digestible maltodextrin.
This supplement was taken 3 times a day. The control supplement was
composed of 6 g digestible maltodextrin.
[0050] A stool sample was taken before the first supplementation,
at the 25th week of gestation. A second stool sample was taken at
the last standardized routine hospital visit before delivery. At
delivery, one cord blood sample was obtained from the placental
umbilical vein. At day 5, day 20 and 6 month after delivery stool
samples were obtained from the neonates.
[0051] Microbial flora of the maternal and neonatal stool samples
were analysed by FISH and real time PCR. Cord blood (CB) was
phenotypically characterized using FACS analysis and in vitro
stimulation assays with mitogens and allergens.
[0052] The following parameters were measured using methods known
in the art:
a) Innate Immune Response
[0053] NKT cells, frequency of monoclonal T cell receptors (i.e.
homogeneous N-region of Va24JaQ TCR, quantification of Toll-like
receptor expressing cells (TLR2, TLR4); granulocytes count. NK-cell
activity
b) Adaptive Immune Response
[0054] Lymphocyte subsets were by four-color-cytometry.
Characterization of the TH1/TH2-related mRNA cytokine and chemokine
receptor patterns (TNF-a, INF-g, IL-4, IL-8, IL10, IL-12, IL-13,
TGF.beta., CD25, CD45RO, CD45RO, CTLA-4, CCR3, CCR4, CCR5, CXCR3)
in non-stimulated cells were determined by Real-time RT PCR and
after whole blood stimulation with ConA/PHA and LPS ex vivo.
Analysis of the ratio of TH1/TH2-cytokine production within
CD4.sup.+ and CD8.sup.+T-cell subsets were determined.
[0055] Placental cortical blood samples were obtained form 28
deliveries and stimulated with ConcanavalinA (ConA)/(PHA),
Lipopolysaccharide (LPS), Staphylococcus enterotoxin B (SEB),
.beta.-lactoglobulin (BLG), major house dust mite allergen from
Dermatophagoides pteronyssinus (Der p1), ovalbumin (Ova), for 24 h.
CB T lymphocytes (50 .mu.L whole blood samples) were stained with
appropriately diluted FITC-, PE-, PC5-, or APC-labeled monoclonal
antibodies against human CD4, CD8, CD45RO, CD45RO, CD25, CD69,
CCR4, CCR5, CD14, CCR1, CCR2, CCR6, CCR7, CCR8, CCR9, CXCR3, CXCR4,
CXCR5, CRTH2, and appropriate isotype controls. After staining,
contaminating erythrocytes were lysed. The percentages of chemokine
receptor (CKR.sup.+) and of CKR.sup.+CD45RA.sup.+ expressing
lymphocytes within the CD4.sup.+ T helper (Th) population, and the
percentages of CKR.sup.+ expressing cells within the
CD4.sup.+CD45RA.sup.+ population were analyzed. The same analyses
procedure was performed within the CD8.sup.+ T cytotoxic (Tc) and
CD8.sup.+CD45RA.sup.+ lymphocyte populations. For CXCR4 the mean
fluorescence intensity (MFI) was determined. CD4.sup.+CD25.sup.high
expression on T regulatory cells was determined.
Whole CB Stimulation Assay
[0056] Heparinized CB was diluted 1 to 5 in RPMI-1640. Aliquots of
1 mL diluted CB were stimulated with concavalin A (Con A 50
.mu.g/mL), beta-lactoglobulin (100 .mu.g/mL BLG), ovalbumin (100
.mu.g/mL OVA), lipopolysaccharid (0.1 .mu.g/mL LPS), staphylococcal
enterotoxin B (0.1 .mu.g/mL SEB), Dermatophagoides pteronyssinus
(10 .mu.g/mL Der p1) or medium alone in a 24-well culture plate.
Cells were incubated at 37.degree. C. in a 5% CO.sub.2-atmosphere
for 24 h or 48 h and supernatants were stored at -80.degree. C. for
cytokine analyses.
Multiplex Array
[0057] The cytokines were quantified in culture supernatants with a
human multiplex, particle-based, flow cytometric assay.
TNF-.alpha., IFN-.gamma., IL-1.beta., IL-2, IL-4, IL-10, GM-CSF,
G-CSF concentrations in the supernatants were directly measured
without further dilution. To determine IL-6, IL-8, MCP-1 and
MIP-1.beta. supernatants were diluted 1:10 in RPMI. The differences
in cytokine expression, as compared to the negative control, were
determined and concentrations were normalized to the number of
lymphocytes (for IL-2) and total leukocytes (for all other
cytokines) obtained from whole blood count (pg/ml/10E3 cells).
[0058] Despite large intra-individual variations in the maternal
microflora composition, the total bacterial load and the median
numbers of cells/ml of bifidobacteria and Lactobacilli did not
differ between both groups before onset of supplementation.
However, upon supplementation with the supplement comprising GOS
and inulin an increased percentage of total bifidobacteria and
lactobacilli in the faecal flora of the pregnant woman was
observed. See Table 1. In maternal stool samples, B. catenulatum,
B. infantis and L. acidophilus were the most frequent species in
both groups before and after supplementation.
[0059] The faeces of the infants from mothers from the experimental
group contained, at day 5 after delivery, 7.4% lactobacilli (n=17,
s.e. 3.0), whereas the faeces of infants of mothers of the control
group, at day 5 after delivery, contained only 2.4% lactobacilli
(n=16, s.e. 0.9).
[0060] Intra-group comparisons of infants showed a significant
increase in the numbers of lactobacilli in the placebo group
[p=0.038; pW] from day 20 to day 182. This effect was confirmed by
FISH analyses [p=0.018; pW]. The percentage of neonates positive
for L. acidophilus increased in the placebo group from day 20 to
day 182 [p=0.016]. This difference was confirmed by qPCR analyses
(p=0.012).
[0061] FACS analysis of cord blood revealed significantly lower
mean fluorescence intensity levels of TLR2.sup.+ on CD14.sup.+
monocytes in the experimental group (p<0.05). The altered CD14+
and TLR2+ expression levels indicate that the neonatal immune
system before delivery is directed towards a faster Th1 response
after delivery.
TABLE-US-00001 TABLE 1 Percentage bifdidobacteria and lactobacilli
in faeces of pregnant women consuming a mixture of GOS and inulin
(experimental) or maltodextrin (control). Experimental Experimental
Control Control Mean (s.e.) % Mean (s.e.) % Mean (s.e.) % Mean
(s.e.) % bifidobacteria.sup.a lactobacilli.sup.a
bifidobacteria.sup.a lactobacilli.sup.a T = 1.sup.b 13.0
(1.7).sup.c 8.6 (2.0) 18.5 (4.1) 5.4 (1.9) T = 2.sup.b 24.1
(2.6).sup.cd 9.9 (1.7) 14.7 (3.5).sup.d 7.8 (1.9)
.sup.aBifidobacteria were quantified with FISH, lactobacilli were
quantified by real time PCR. .sup.bt = 1 in 25.sup.th week of
gestation, before onset of supplementation, t = 2 is at the last
hospital visit before delivery. .sup.cStatistically significant
difference, p < 0.05, between t = 1 and t = 2
.sup.dStatistically significant difference, p < 0.05, between
experimental and control group.
[0062] To test if maternal prebiotic supplementation affected fetal
immunity, a comprehensive phenotypical lymphocyte subset analyses
was performed. CB CD4.sup.+ T helper (Th) cells and CD8.sup.+ T
cytotoxic (Tc) cells predominantly expressed the naive
(CD45RA.sup.+) phenotype (>84%) while less than 13% expressed
CD45RO.sup.+. The ratio of CD4.sup.+/CD8.sup.+ as well as the
percentages of CD4.sup.+CD45RA.sup.+, CD8.sup.+CD45RA.sup.+,
CD4.sup.+CD45RO.sup.+ and CD4.sup.+CD45RO.sup.+ did not differ
significantly between the prebiotic and the placebo groups. In
contrast to the high frequency of CCR7.sup.+ (>87.3%) cells, all
other CKRs were expressed at a very low frequency on both CD4.sup.+
and CD8.sup.+ cells. The MFI of CXCR4 was high on both CD4.sup.+
and CD8.sup.+ subsets, while CXCR3 was only expressed at a high
frequency on CD8.sup.+ cells. The analyses of the frequency of
CKR.sup.+ lymphocytes within CD4.sup.+, CD8.sup.+,
CD4.sup.+CD45RA.sup.+ and CD8.sup.+CD45RA.sup.+ CB T cell subsets,
as well as the Th1/Th2 ratio (CXCR3/CCR4 and CCR5/CCR4) within the
different CD4.sup.+ and CD8.sup.+ subsets, did not differ
significantly between the two groups. The early activation marker
CD69 as well as the late activation marker CD25 were expressed at
similar levels on the different CD4.sup.+ subsets in both groups
(data not shown). No difference in the frequency of CD25.sup.high
expressing CD4.sup.+ T cell subsets was observed (median: 1.5
versus 1.4; p=0.683).
[0063] The comparative analyses of leukocyte-derived cytokine
profile production by whole CB cultures stimulated with mitogen and
allergens for 24 h revealed typical patterns of cytokine
expression. No significant differences between the two
supplementation groups were observed, when analyzing the cytokine
concentrations induced by different stimuli (apart from G-CSF
(p<0.03 for Der p1, BLG and OVA).
[0064] These results indicate that supplementation with water
soluble non-digestible saccharides resulted in increased percentage
of bifidobacteria and lactobacilli in the gut microflora of
pregnant women. This results in an improved inoculation of the
intestine of the infant during birth and colonisation after birth
with lactobacilli and/or bifidobacteria. Especially the presence of
lactobacilli very soon after birth is of importance, since
lactobacilli have a high capacity of acidification of the colon,
thereby making the intestinal environment subsequently more
suitable for bifidobacteria and less suitable for enteral
pathogens. Furthermore, these results indicate that this
supplementation improves the immune system of the unborn child and
infant during and after birth.
Example 2
[0065] 100 ml of a liquid milk-based composition packed in a 100 ml
bottle comprising: [0066] 8.1 g transgalacto-oligosaccharide,
(originating from Vivinal GOS) [0067] 0.9 g fructopolysaccharide
(originating from RaftilineHP) [0068] 750 mg fish oil and evening
primrose oil, comprising [0069] 300 mgDHA [0070] 42 mg EPA [0071]
8.4 mgAA [0072] 8.4 mg docosapentaenoic acid (DPA) [0073] 15 mg
gamma linoleic acid (GLA)
* * * * *