U.S. patent application number 16/563394 was filed with the patent office on 2019-12-26 for composition with non-digestible oligosaccharides for attenuating nasal epithelial inflammation.
The applicant listed for this patent is Johan GARSSEN, Caroline SCHLUMPRECHT, Claudia TRAIDL-HOFFMANN, Jeroen VAN BERGENHENEGOUWEN. Invention is credited to Johan GARSSEN, Caroline SCHLUMPRECHT, Claudia TRAIDL-HOFFMANN, Jeroen VAN BERGENHENEGOUWEN.
Application Number | 20190388445 16/563394 |
Document ID | / |
Family ID | 58265834 |
Filed Date | 2019-12-26 |
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United States Patent
Application |
20190388445 |
Kind Code |
A1 |
VAN BERGENHENEGOUWEN; Jeroen ;
et al. |
December 26, 2019 |
COMPOSITION WITH NON-DIGESTIBLE OLIGOSACCHARIDES FOR ATTENUATING
NASAL EPITHELIAL INFLAMMATION
Abstract
The present invention relates to the use of non-digestible
oligosaccharides for the direct attenuation of nasal epithelial
inflammation, in particular for non-allergic rhinitis, in
particular for use in infants.
Inventors: |
VAN BERGENHENEGOUWEN; Jeroen;
(Utrecht, NL) ; GARSSEN; Johan; (Utrecht, NL)
; TRAIDL-HOFFMANN; Claudia; (Augsburg, DE) ;
SCHLUMPRECHT; Caroline; (Augsburg, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
VAN BERGENHENEGOUWEN; Jeroen
GARSSEN; Johan
TRAIDL-HOFFMANN; Claudia
SCHLUMPRECHT; Caroline |
Utrecht
Utrecht
Augsburg
Augsburg |
|
NL
NL
DE
DE |
|
|
Family ID: |
58265834 |
Appl. No.: |
16/563394 |
Filed: |
September 6, 2019 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
PCT/EP2018/055708 |
Mar 8, 2018 |
|
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16563394 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A23V 2002/00 20130101;
A23V 2002/00 20130101; A23V 2200/304 20130101; A61K 2300/00
20130101; A23L 33/125 20160801; A61K 47/26 20130101; A23L 33/40
20160801; A61K 31/733 20130101; A61K 31/733 20130101; A61K 9/0043
20130101; A61K 31/702 20130101; A61K 2300/00 20130101; A61K 9/0095
20130101; A61P 11/02 20180101; A61K 31/702 20130101; A23V 2250/284
20130101 |
International
Class: |
A61K 31/702 20060101
A61K031/702; A61K 9/00 20060101 A61K009/00; A61P 11/02 20060101
A61P011/02 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 8, 2017 |
EP |
17159857.6 |
Claims
1. A method of treating and/or preventing non-allergic rhinitis in
a human subject, comprising administering to a subject in need
thereof a composition comprising non-digestible
oligosaccharides.
2. The method according to claim 1, wherein the non-digestible
oligosaccharides comprise galacto-oligosaccharides and/or
fructo-oligosaccharides.
3. The method according to claim 2, wherein the non-digestible
oligosaccharides comprise galacto-oligosaccharides and
fructo-oligosaccharides.
4. The method according to claim 1, wherein the human subject has
an age of 0 to 36 months.
5. The method according to claim 4, wherein the human subject is an
infant.
6. The method according to claim 1, wherein the composition is a
nutritional composition.
7. The method according to claim 6, wherein the nutritional
composition is an infant formula or follow on formula.
8. The method according to claim 1, wherein the non-allergic
rhinitis is a chronic non-allergic rhinitis.
9. The method according to claim 1, wherein the non-allergic
rhinitis is a non-infectious non-allergic rhinitis.
10. The method according to claim 1, wherein the administering
comprises directly contacting the composition with nasal epithelial
cells.
11. The method according to claim 1, wherein the human subject is
exposed to environmental air pollutants.
12. The method according to claim 1, wherein the composition does
not comprise Lactobacillus rhamnosus.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application is a bypass continuation of
International Application No. PCT/EP2018/055708 filed Mar. 8, 2018,
which claims priority to European Application No. 17159857.6 filed
Mar. 8, 2017, the contents of which are hereby incorporated by
reference in their entirety.
FIELD OF THE INVENTION
[0002] The invention relates to the use of non-digestible
oligosaccharides for nasal inflammation.
BACKGROUND OF THE INVENTION
[0003] Rhinitis is generally considered to be chronic or acute
inflammation of the mucous membrane of the nose. This inflammation
results in the production of excessive amounts of mucus, resulting
in the primary symptom of rhinitis, nasal dripping or "runny nose",
as well as nasal congestion, rhinoconjunctivitis (inflammation of
conjunctiva) and post-nasal drip.
[0004] Rhinitis is commonly categorised into three types:
infectious rhinitis, including acute and chronic bacterial or viral
infections; allergic rhinitis, commonly triggered by inhaled
allergens present in for example pollen, mold, animal dander, and
dust mite, and non-infectious non-allergic rhinitis. This latter
type of rhinitis can be chronic and can be caused by non-allergic
triggers in the environment such as smells, fumes, smoke, dusts,
car exhausts, chlorine, cigarette smoke, cleaning solutions, ozone,
smog, fine particles and temperature changes. Environmental
irritants are common triggers of non-allergic rhinitis.
[0005] A blocked, "stuffy" nose is typically not considered to be a
serious condition. However for infants and children the impact may
be higher than for adults. It has a considerable negative impact on
the quality of life of both the parent and child. Infants with
blocked noses struggle to breathe while being milk-fed, and young
children are unable to blow their noses properly to ease their
discomfort. Infants and children with a blocked nose have
difficulty sleeping at night too. Parents often seek help on how to
clear their child's nose. Furthermore, young infants are not able
to mouth breath properly and can only do so by crying. Rhinitis in
infants and children has been associated with sleeping problems,
ear conditions, and even learning problems.
[0006] Current treatment regimes are directed to minimising
exposure to the triggers of rhinitis and suppressing symptoms.
Saltwater sprays, rinses or steam may be effective to remove dust,
secretions and allergenic molecules from the mucosa, and so
minimize exposure.
[0007] WO 2011/099875 discloses the use of Lactobacillus rhamnosus
HN001 or derivatives thereof to treat or prevent rhinitis. However,
the effect of non-digestible oligosaccharides is not tested.
[0008] US 2009/0280099 discloses nasopharyngeal inoculate of
probiotics for treatment of respiratory infections. However, the
effect of non-digestible oligosaccharides is not tested.
[0009] WO 2005/039597 relates to a method for enhancing the immune
system and the treatment and/or prevention of immune system related
disorders in a mammal, particularly newborns, said method
comprising the administration of acid oligosaccharide and neutral
oligosaccharide. In particular an effect on allergy and atopic
diseases is aimed for.
SUMMARY OF THE INVENTION
[0010] The investigators surprisingly found that non-digestible
oligosaccharides had a direct effect on down-regulation of the
inflammatory response of human nasal epithelial cells from healthy
non-atopic subjects. Results revealed that the presence of the
non-digestible oligosaccharides directly decreased the secretion of
pro-inflammatory chemokines such as IP-10, CCL5 and CCL20 in
epithelial cells and the anti-inflammatory effects were at least as
high, or even higher, in nasal epithelial cells from non-atopic
subjects than those of in nasal epithelial cells of atopic, i.e.
allergic, subjects. Furthermore, the effects were higher compared
to those of Lactobacillus rhamnosus. In particular in infants and
young children preventing and/or treatment of non-allergic
rhinitis, will be advantageous. The present invention thus is
directed to compositions comprising non-digestible
oligosaccharides, in particular galacto- and/or
fructo-oligosaccharides, for preventing and treating non-allergic
rhinitis.
DETAILED DESCRIPTION OF THE INVENTION
[0011] The present invention concerns a method for treating and/or
preventing non-allergic rhinitis in a human subject, comprising
administering to said human subject a composition comprising
non-digestible oligosaccharides.
[0012] The present method can also be referred to as a non-medical
method for treating and/or preventing non-allergic rhinitis in a
human subject.
[0013] For some jurisdictions, the invention can also be worded as
the use of non-digestible oligosaccharides in the manufacture of a
composition for treating and/or preventing non-allergic rhinitis in
a human subject.
[0014] For some jurisdictions, the invention can also be worded as
a composition comprising non-digestible oligosaccharides for use in
treating and/or preventing non-allergic rhinitis in a human
subject.
[0015] Non-Digestible Oligosaccharides
[0016] The composition in the present method or use comprises
non-digestible oligosaccharide and preferably comprises at least
two non-digestible oligosaccharides, in particular two different
sources of non-digestible oligosaccharide. The non-digestible
oligosaccharides were found to directly decrease the inflammatory
response in nasal epithelial cells, in particular of non-atopic
subjects.
[0017] The term "oligosaccharide" as used herein refers to
saccharides with a degree of polymerization (DP) of 2 to 250,
preferably a DP 2 to 100, more preferably 2 to 60, even more
preferably 2 to 10. If oligosaccharide with a DP of 2 to 100 is
included in the composition in the present method or use, this
results in compositions that may contain oligosaccharides with a DP
of 2 to 5, a DP of 50 to 70 and a DP of 7 to 60. The term
"non-digestible oligosaccharide" as used in the present invention
refers to oligosaccharides which are not digested in the intestine
by the action of acids or digestive enzymes present in the human
upper digestive tract, e.g. small intestine and stomach, but which
are preferably fermented by the human intestinal microbiota. For
example, sucrose, lactose, maltose and maltodextrins are considered
digestible.
[0018] Preferably the present non-digestible oligosaccharide is
soluble. The term "soluble" as used herein, when having reference
to a polysaccharide, fibre or oligosaccharide, means that the
substance is at least soluble according to the method described by
L. Prosky et al., J. Assoc. Off. Anal. Chem. 71, 1017-1023
(1988).
[0019] In one embodiment, the non-digestible oligosaccharide
included in the compositions in the method or use according to the
present invention is preferably selected from the group consisting
of fructo-oligosaccharides (such as inulin), non-digestible
dextrins, galacto-oligosaccharides (such as
transgalacto-oligosaccharides), xylo-oligosaccharides,
arabino-oligosaccharides, arabinogalacto-oligosaccharides,
gluco-oligosaccharides, gentio-oligosaccharides,
glucomanno-oligosaccharides, galactomanno-oligosaccharides,
mannan-oligosaccharides, isomalto-oligosaccharides,
nigero-oligosaccharides, chito-oligosaccharides, soy
oligosaccharides, uronic acid oligosaccharides,
sialyloligosaccharides, sialyllactose, lactosialylterasaccharide
a,b,c, disialyllactoNtetraose, sialyl-lactoNhexaose,
sialyl-lactoNhexaose, fucosylated-oligosaccharides, (such as
(un)sulphated fucoidan oligosaccharides, fucosyllactose,
difucosyllactose, lacto-N-fucopenatose, lacto-N-neofucopenaose,
lacto-N-difucosyl-hexaose) and mixtures thereof,
[0020] In one embodiment, the non-digestible oligosaccharide
included in the compositions in the method or use according to the
present invention preferably include a mixture of non-digestible
oligosaccharides. The non-digestible oligosaccharide is preferably
selected from the group consisting of fructo-oligosaccharide and
galacto-oligosaccharide, even more preferably is selected from the
group consisting of transgalacto-oligosaccharide and inulin, most
preferably is transgalacto-oligosaccharide.
[0021] In one embodiment, the non-digestible oligosaccharide is
preferably selected from the group consisting of
beta-galacto-oligosaccharide, alpha-galacto-oligosaccharide and
galactan. According to a more preferred embodiment the
non-digestible oligosaccharide is beta-galacto-oligosaccharide.
Preferably the non-digestible oligosaccharide comprises
galacto-oligosaccharide with beta-(1,4), beta-(1,3) and/or
beta-(1,6) glycosidic bonds and a terminal glucose.
Transgalacto-oligosaccharide is for example available under the
trade name Vivinal.RTM.GOS (Domo FrieslandCampina Ingredients),
Bi2muno (Clasado), Cup-oligo (Nissin Sugar) and Oligomate55
(Yakult). These oligosaccharides are thought to have a superior
effect in decreasing the inflammatory response in nasal epithelial
cells.
[0022] In a preferred embodiment, the galacto-oligosaccharide
comprised in the composition for the method or use according to the
present invention has an average DP in the range of 2-10,
preferably 2-7, preferably 3-6.
[0023] In one embodiment, the non-digestible oligosaccharide
preferably comprises fructo-oligosaccharide. A
fructo-oligosaccharide may in other context have names like
fructopolysaccharide, oligofructose, polyfructose, polyfructan,
inulin, levan and fructan and may refer to oligosaccharides
comprising beat-linked fructose units, which are preferably linked
by beta-(2,1) and/or beta-(2,6) glycosidic linkages, and preferably
have a DP between 2 and 200. Preferably, the fructo-oligosaccharide
contains a terminal beta-(2,1) glycosidic linked glucose.
[0024] Preferably, the fructo-oligosaccharide contains at least 7
beta-linked fructose units. In a further preferred embodiment
inulin is used. Inulin is a type of fructo-oligosaccharide wherein
at least 75% of the glycosidic linkages are beta-(2,1) linkages.
Typically, inulin has an average chain length between 8 and 60
monosaccharide units. A suitable fructo-oligosaccharide for use in
the compositions according to the method or use of the present
invention is commercially available under the trade name
Raftiline.RTM.HP (Orafti). Other suitable sources are Raftilose
(Orafti), Fibrulose and Fibruline (Cosucra) and Frutafit and
Frutalose (Sensus).
[0025] In a preferred embodiment, the fructo-oligosaccharide
comprised in the composition for the method or use according to the
present invention has an average DP in the range of 7-100,
preferably 11-60, preferably 20-50.
[0026] In one embodiment, the composition in the present method or
use comprises a mixture of galacto-oligosaccharide and
fructo-oligosaccharide. Preferably the mixture of
galacto-oligosaccharide and fructo-oligosaccharide is present in a
weight ratio of from 1/99 to 99/1, more preferably from 1/19 to
19/1, more preferably from 1/1 to 19/1, more preferably from 2/1 to
15/1, more preferably from 5/1 to 12/1, even more preferably from
8/1 to 10/1, even more preferably in a ratio of about 9/1. This
weight ratio is particularly advantageous when
galacto-oligosaccharide has a low average DP and
fructo-oligosaccharide has a relatively high DP. Most preferred is
a mixture of galacto-oligosaccharide with an average DP below 10,
preferably below 6, preferably in the range of 2-10, preferably in
the range of 2-7, preferably in the range of 3-6, and
fructo-oligosaccharide with an average DP above 7, preferably above
11, even more preferably above 20, preferably in the range of
7-100, preferably in the range of 11-60, preferably in the range of
20-50. Such a mixture is thought to have further improved effects
on decreasing the inflammatory response in nasal epithelial
cells.
[0027] In one embodiment, the composition in the present method or
use comprises a mixture of short chain fructo-oligosaccharide and
long chain fructo-oligosaccharide. Preferably the mixture of short
chain fructo-oligosaccharide and long chain fructo-oligosaccharide
is present in a weight ratio of from 1/99 to 99/1, more preferably
from 1/19 to 19/1, even more preferably from 1/10 to 19/1, more
preferably from 1/5 to 15/1, more preferably from 1/1 to 10/1.
Preferred is a mixture of short chain fructo-oligosaccharide with
an average DP below 10, preferably below 6, preferably in the range
of 2-10, preferably in the range of 2-7, preferably in the range of
3-6, and fructo-oligosaccharide with an average DP above 7,
preferably above 11, even more preferably above 20, preferably in
the range of 7-100, preferably in the range of 11-60, preferably in
the range of 20-50.
[0028] Preferably the composition in the present method or use
comprises 2.5 to 20 wt % total non-digestible oligosaccharide, more
preferably 2.5 to 15 wt %, even more preferably 3.0 to 10 wt %,
most preferably 5.0 to 7.5 wt %, based on total dry weight of the
composition. When in liquid form, the composition in the present
method or use preferably comprises 0.35 to 2.5 wt % total
non-digestible oligosaccharide, more preferably 0.35 to 2.0 wt %,
even more preferably 0.4 to 1.5 wt %, based on 100 ml of the
composition. A lower amount of non-digestible oligosaccharide will
be less effective in decreasing the inflammatory response in nasal
epithelial cells, whereas a too high amount will result in
side-effects of bloating and abdominal discomfort after ingestion
by the human subject.
[0029] Composition
[0030] In one embodiment, the composition in the method or use
according to the present invention is a nutritional composition.
Preferably the composition is a nutritional composition that is
suitable for administration to infants. In one embodiment, the
composition in the method or use according to the present invention
is a pharmaceutical composition, and preferably is a pharmaceutical
composition that is suitable for administration to infants. In one
embodiment the composition is in a form that is suitable for
topical administration in the nasal cavity of a human subject,
preferably the composition is in the form of administration as a
spray or as drops, or is in the form of a spray or drops. In one
embodiment the composition is administered topically, more
preferably topically in the nasal cavity. In one embodiment the
composition is enterally administered, more preferably is orally
administered. As nasal cavity, oral cavity and throat are in close
proximity with each other, in particular in infants and young
children, more particular in infants given their different anatomy,
it is thought that oral administration will also result in a direct
contact of the orally administered composition. This direct contact
with the nasal epithelium is in infants further present due to
burping, choking and coughing while feeding, and uncomplicated
(nasal) regurgitation, even in otherwise healthy infants.
[0031] The composition in the method or use according to the
present invention therefore preferably is a nutritional
composition, more preferably an infant formula, follow on formula,
or young child formula. The present nutritional composition can be
advantageously applied as a complete nutrition for infants.
Preferably the present nutritional composition is an infant
formula. An infant formula is defined as a formula for use in
infants and can for example be a starter formula, intended for
infants of 0 to 6 or 0 to 4 months of age. A follow on formula is
intended for infants of 4 or 6 months to 12 months of age. A
toddler or growing up milk or formula is intended for children of
12 to 36 months of age. The present composition preferably
comprises a lipid component, protein component and carbohydrate
component and is preferably administered in liquid form. The
present nutritional composition may also be in the form of a dry
food, preferably in the form of a powder which is accompanied with
instructions as to mix said dry food, preferably powder, with a
suitable liquid, preferably water. The nutritional composition used
according to the invention preferably comprises other fractions,
such as vitamins, minerals, trace elements and other micronutrients
in order to make it a complete nutritional composition. Preferably
infant formulas comprise vitamins, minerals, trace elements and
other micronutrients according to international directives.
[0032] The nutritional composition in the method or use according
to the present invention preferably comprises lipid, protein and
digestible carbohydrate invention wherein the lipid provides 5 to
50% of the total calories, the protein provides 5 to 50% of the
total calories, and the digestible carbohydrate provides 15 to 90%
of the total calories. Preferably, in the nutritional composition
the lipid provides 35 to 50% of the total calories, the protein
provides 7.5 to 12.5% of the total calories, and the digestible
carbohydrate provides 40 to 55% of the total calories. The
nutritional composition in the method or use according to the
present invention is not human milk.
[0033] In order to meet the caloric requirements of an infant or
toddler, the nutritional composition preferably comprises 45 to 200
kcal/100 ml liquid. For infants the nutritional composition has
more preferably 60 to 90 kcal/100 ml liquid, even more preferably
65 to 75 kcal/100 ml liquid. This caloric density ensures an
optimal ratio between water and calorie consumption. For toddlers,
human subjects with an age between 12 and 36 months, the
nutritional composition more preferably has a caloric density
between 45 and 65, even more preferably between 50 and 60 kcal/100
ml. The osmolarity of the present composition is preferably between
150 and 420 mOsmol/l, more preferably 260 to 320 mOsmol/l. The low
osmolarity aims to further reduce the gastrointestinal stress. The
present nutritional composition, when in liquid form, preferably
has a viscosity between 1 and 60 mPas, preferably between 1 and 20
mPas, more preferably between 1 and 10 mPas, most preferably
between 1 and 6 mPas. The low viscosity ensures a proper
administration of the liquid, and low viscosity will further
facilitate contact with nasal epithelium in infants. Also this
viscosity closely resembles the viscosity of human milk.
Furthermore, a low viscosity results in a normal gastric emptying
and a better energy intake, which is essential for infants which
need the energy for optimal growth and development. The nutritional
composition is preferably prepared by admixing a powdered
composition with water. Normally infant formula is prepared in such
a way. The composition for the method or use according to the
present invention thus also relates to a packaged power composition
wherein said package is provided with instructions to admix the
powder with a suitable amount of liquid, thereby resulting in a
liquid composition with a viscosity between 1 and 60 mPas. The
viscosity of the liquid is determined using a Physica Rheometer MCR
300 (Physica Messtechnik GmbH, Ostfilden, Germany) at a shear rate
of 95 s.sup.-1 at 20.degree. C.
[0034] In one embodiment the composition in the method or use
according to the present invention is a pharmaceutical composition,
and preferably is a pharmaceutical composition that is suitable for
administration to adults. Preferably the composition is applied
nasally, preferably the composition is applied nasally in an adult
human subject.
[0035] As the effect of tested Lactobacillus rhamsosus was less
than that of the non-digestible oligosaccharides, and a combination
of Lactobacillus rhamnosus and non-digestible oligosaccharides
showed no improved effect on attenuating the inflammatory response
in nasal epithelial cells, preferably the composition does not
comprises Lactobacillus rhamnosus. Preferably the composition does
not comprise Lactobacillus. The present finding that non-digestible
oligosaccharides alone are as effective or even more effective than
probiotic Lactobacillus rhamnosus has also further advantages:
Administration of non-digestible oligosaccharides is easier to dose
than administration of Lactobacillus. Furthermore such compositions
with non-digestible oligosaccharides and without Lactobacillus
rhamnosus are safer, require less careful handling to keep the
active ingredient active, and have an improved shelf life. In one
embodiment, the composition in the method or use according to the
present invention does not comprise Lactobacillus rhamsosus HN001.
In one embodiment, the composition in the method or use according
to the present invention does not comprise Aspergillus, Bacillus,
Bacteroides, Bifidobacterium, Lactobacillus, Leuconostoc,
Pediococcus, Propionibacterium, Saccharomyces, and Enterococcus. In
one embodiment, the composition in the method or use according to
the present invention does not comprise a probiotic.
[0036] Application
[0037] In the context of the present invention, `prevention` of a
disease or certain disorder also means `reduction of the risk` of a
disease or certain disorder and also means `treatment of a person
at risk` of said disease or said certain disorder.
[0038] The inventors surprisingly found that direct application of
non-digestible oligosaccharides to human nasal epithelial cells
(HNECs) of non-atopic subjects attenuated the inflammatory response
to a higher extent than from non-atopic subjects and to a higher
extent than of atopic subjects. Non-digestible oligosaccharides
also advantageously improve the barrier function of the nasal
epithelial cells, e.g as measured as an increase or smaller
decrease of the transepithelial electrical resistance (TEER). This
finding is indicative for a treatment and/or prevention of
non-atopic rhinitis, as firstly it can be expected that an indirect
effect of non-digestible oligosaccharides via improving of the
immune system involving the gastrointestinal tract, e.g. by a
systemic increase the balance of Th1/Th2 response, will have no or
less effect on non-allergic rhinitis, and secondly the effects
observed were much more pronounced in HNECs of non-atopic, i.e.
non-allergic, subjects.
[0039] As the impact of rhinitis is also higher in infants and
young children, in particular infants, when compared to adults, the
current invention is preferably advantageously used for infants and
young children, more preferably infants. Infants with blocked noses
struggle to breathe while being milk-fed, and young children are
unable to blow their noses properly to ease their discomfort.
Furthermore, young infants are not able to mouth breath properly
and can only do so by crying. Rhinitis in infants and children has
been associated with sleeping problems, ear conditions, and even
learning problems.
[0040] In one embodiment, the composition in the present invention
is used for preventing and/or treatment non-allergic rhinitis in a
human subject, more preferably a human subject with an age of 0 to
36 months, more preferably of 0 to 18 months, even more preferably
an infant with an age of 12 months of age or below, even more
preferably an infant with an age of 0 to 6 months. Preferably the
composition is further used for providing nutrition to said human
subject, more preferably complete nutrition. In a preferred
embodiment, the method or use according to the present invention is
for term infants, preferably for healthy term infants.
[0041] Preferably the non-allergic rhinitis is non-infectious
non-allergic rhinitis (NINAR). Preferably the non-allergic rhinitis
is chronic non-allergic rhinitis, as chronic rhinitis has the most
impact on health. Preferably the treatment and/or prevention of
non-allergic rhinitis is by a direct contact of the non-digestible
oligosaccharides with the nasal epithelial cells. This has the
advantage that the treatment and/or prevention is locally, at the
location that is affected only. In one embodiment, non-allergic
rhinitis can also be referred to as inflammatory non-allergic
rhinitis or as inflammation of the nasal epithelium.
[0042] In one embodiment, the non-allergic rhinitis is treated or
prevented in a human subject, more preferably a child or an infant,
that is exposed to environmental air pollutants. Such human
subjects are in particular at risk of having non-allergic rhinitis,
in particular chronic non-atopic rhinitis and/or non-infectious
non-atopic rhinitis.
[0043] 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. Wt % means weight percentage.
DESCRIPTION OF THE FIGURES
[0044] FIGS. 1A and 1B: CCL5 release of human primary nasal
epithelial cells from non-atopic subjects (A) or atopic subjects
(B) stimulated with IFN-.gamma./TNF-.alpha. and scGOS/IcFOS, L.
rhamnosus GG or their combination, or a control. CCL5 was
normalized on IFN-.gamma./TNF-.alpha..
[0045] FIGS. 2A and 2B: IP-10 release of human primary nasal
epithelial cells from non-atopic subjects (A) or atopic subjects
(B) stimulated with IFN-.gamma./TNF-.alpha. and scGOS/IcFOS, IMS1,
L. rhamnosus or their combination. IP-10 was normalized on
IFN-.gamma./TNF-.alpha..
EXAMPLES
Example 1: Effects of Prebiotics and Probiotics on
IFN-.gamma./TNF-.alpha. Stimulated Nasal Epithelial Cells
[0046] Since long the use of human nasal epithelial cells has been
known as a suitable in vitro model system, see for example Schmidt
et al., Advanced Drug Delivery Reviews, 1998, 29:51-79. Human
primary nasal epithelial cells (HNECs) were isolated from
non-atopic (NA) subjects, grown and frozen in liquid nitrogen for
further use. To exert the experiments in monolayer, human primary
epithelial cells (HNECs) were thawed and seeded in Airway
Epithelial Cell Growth Medium (AEGM) with supplements (PromoCell,
C-21060) in 24 wells plates. The cells were incubated at 37.degree.
C. at 5 CO.sub.2 for 5-6 days until they reached 80% confluence. On
the morning of the experiment the mixture short chain
galacto-oligosaccharides/long chain fructo-oligosaccharides
(scGOS/IcFOS) was freshly prepared. Moreover probiotic bacteria,
Lactobacillus rhamnosus LW744 and cytokines were thawed and diluted
to the working concentration. scGOS/IcFOS was mixed in a 9/1 w/w
ratio. VivinalGOS was the source of scGOS, RaftilinHP was the
source of IcFOS. The working concentration of scGOS/IcFOS was 0.5%
w/v, of IFN-.gamma. was 300 IU, of TNF-.alpha. was 10 ng/ml, of
L.rhamnosus rhamnosus LW744 was 10.sup.4 cfu. The Toll-like
receptor 3 (TLR3) ligand polyinosinic-polycytidylic acid (Poly
I:C), a stimulator of the inflammatory response, was thawed and
diluted to the working concentration. The working concentration of
Poly I:C was 10 .mu.g/ml. All stimulants and conditions were
applied simultaneously and incubated with the cells for 24 h at
37.degree. C., 5% CO.sub.2. HNECs from NA subjects were incubated
in AEGM (negative control) or stimulated with IFN-.gamma. and
TNF-.alpha. (positive control) with or without scGOS/IcFOS, L.
rhamnosus LW744 and their combinations in different concentrations
for 24 h at 37.degree. C., 5% CO.sub.2. After 24 h, supernatants
were taken and frozen at -80.degree. C. for further use.
Subsequently, supernatants were used to perform CCL5, IP-10, and
CCL20 ELISAs in cell free supernatants. Results of three
independent experiments are presented as mean of normalized
levels.+-.SEM. Pg/ml levels were normalized on
IFN-.gamma./TNF-.alpha. control. Significant difference between
stimulation conditions is presented by *, p<0.05, one-way ANOVA
with Bonferroni correction for multiple comparisons.
[0047] Results:
[0048] The highest and significant reduction in
IFN-.gamma./TNF-.alpha. stimulated CCL5 release is observed when
the non-digestible oligosaccharide mixture scGOS/IcFOS (GF) was
added to HNEC cells of non-atopic subjects. See FIG. 1A. Addition
of Lactobacillus rhamnosus (L rham) was less effective and the
combination of scGOS/IcFOS and Lactobacillus rhamnosus did not
perform better that scGOS/IcFOS alone. In HNECs from atopic
subjects also a reduction was observed when scGOS/IcFOS was added,
but not to a higher extent, see FIG. 1B.
[0049] CCL5 or RANTES is a chemotaxis-inducing chemokine playing an
active role in attracting circulating lymphocytes into inflammatory
sites. Reduction of IFN-.gamma./TNF-.alpha.-induced CCL5 release is
thus indicative for a reduced nasal inflammatory response.
[0050] Similarly, in HNECS from non-atopic subjects a significant
reduction in IP-10 release was observed, for Lactobacillus as well
as scGOS/IcFOS, but the highest suppression was observed with
scGOS/IcFOS, see FIG. 2A. The combination of scGOS/IcFOS and
Lactobacillus rhamnosus did not perform better that scGOS/IcFOS
alone. In HNECs from atopic subjects the suppression of induced
IP-10 release by scGOS/IcFOS was observed, but to a lower extent,
see FIG. 2B, than when compared to HNEC of non-atopic subjects.
[0051] IP-10, also known as CXCL10 or small-inducible cytokine B10,
has been attributed to several roles in inflammatory responses,
such as chemo-attraction for monocytes/macrophages, T cells, NK
cells, and dendritic cells, promotion of T cell adhesion to
endothelial cells, antitumor activity, and inhibition of bone
marrow colony formation and angiogenesis. Reduction of
IFN-.gamma./TNF-.alpha. induced IP-10 release is thus indicative
for a reduced nasal inflammatory response.
[0052] Likewise when comparing the inflammatory response in HNEC of
non-atopic subjects as well as HNEC form atopic subjects, a
reduction in Poly I:C induced release of CCL20 release was observed
for scGOS/IcFOS, in the HNEC obtained from non-atopic subjects, but
not in HNEC of atopic subjects, data not shown. CCL20, also known
as LARC or MIP3A, is a chemokine and chemotactic factor that
strongly attracts lymphocytes, dendritic cells and weakly attracts
neutrophils to sites of inflammation. Reduction of CCL20 release is
thus indicative for a reduced nasal inflammatory response.
[0053] These results are indicative of an attenuation of the
inflammatory response in nasal epithelium cells, in particular of
non-allergic human subjects, by non-digestible oligosaccharides, in
particular galacto- and/or fructo-oligosaccharides.
* * * * *