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.

Claims

1. A method of treating non-allergic rhinitis in a human subject, comprising administering to a subject in need thereof a composition comprising non-digestible oligosaccharides, wherein the human subject is an infant and wherein the non-allergic rhinitis is a non-infectious non-allergic rhinitis and wherein the non-digestible oligosaccharides comprise as active ingredients galacto-oligosaccharides and fructo-oligosaccharides.

2. The method according to claim 1, wherein the human subject has an age of 0 to 36 months.

3. The method according to claim 1, wherein the composition is a nutritional composition.

4. The method according to claim 3, wherein the nutritional composition is an infant formula or follow on formula.

5. The method according to claim 1, wherein the non-allergic rhinitis is a chronic non-allergic rhinitis.

6. The method according to claim 1, wherein the administering comprises directly contacting the composition with nasal epithelial cells.

7. The method according to claim 1, wherein the human subject is exposed to environmental air pollutants.

8. The method according to claim 1, wherein the composition does not comprise Lactobacillus rhamnosus.

Description

DESCRIPTION OF THE FIGURES

[0041] FIGS. 1A and 1B: CCL5 release of human primary nasal epithelial cells from non-atopic subjects (A) or atopic subjects (B) stimulated with IFN-65 /TNF-α and scGOS/IcFOS, L. rhamnosus GG or their combination, or a control. CCL5 was normalized on IFN-γ/TNF-α.

[0042] 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-γ/TNF-α and scGOS/IcFOS, IMS1, L. rhamnosus or their combination. IP-10 was normalized on IFN-β/TNF-α.

EXAMPLES

Example 1: Effects of Prebiotics and Probiotics on IFN-γ/TNF-α Stimulated Nasal Epithelial Cells

[0043] 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° 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-γ was 300 IU, of TNF-α 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 μg/ml. All stimulants and conditions were applied simultaneously and incubated with the cells for 24 h at 37° C., 5% CO.sub.2. HNECs from NA subjects were incubated in AEGM (negative control) or stimulated with IFN-γ and TNF-α (positive control) with or without scGOS/IcFOS, L. rhamnosus LW744 and their combinations in different concentrations for 24 h at 37° C., 5% CO.sub.2. After 24 h, supernatants were taken and frozen at −80° 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-γ/TNF-α control. Significant difference between stimulation conditions is presented by *, p<0.05, one-way ANOVA with Bonferroni correction for multiple comparisons.

Results

[0044] The highest and significant reduction in IFN-γ/TNF-α 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.

[0045] CCL5 or RANTES is a chemotaxis-inducing chemokine playing an active role in attracting circulating lymphocytes into inflammatory sites. Reduction of IFN-γ/TNF-α-induced CCL5 release is thus indicative for a reduced nasal inflammatory response.

[0046] 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.

[0047] 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-γ/TNF-α induced IP-10 release is thus indicative for a reduced nasal inflammatory response.

[0048] 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.

[0049] 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.