Prebiotic compounds or compositions for use in treatment of Sjogren's syndrome

Technical field of the invention

The present invention relates to hitherto unknown uses of prebiotic compounds or compositions, e.g. to reduce immune cell infiltrations of salivary glands in Sjogren's syndrome patients. In particular, the present invention relates to specifically administered xylooligosaccharide (XOS) containing compounds or compositions for use in treatment of Sjogren's syndrome. Background of the invention

Sjogren's syndrome is among the most common autoimmune diseases in the Western world. Sjogren's syndrome is an autoimmune disease in which the moisture-producing glands are infiltrated by autoreactive immune cells resulting in oral and ocular dryness. Hitherto, treatment for Sjogren's syndrome has largely been based on treatment of symptoms (e.g, lotion for dry skin, artificial tears for dry eyes). For example, commercial drugs like Rituximab have shown promise in the treatment of severe extraglandular manifestations (e.g., vasculitis, cryoglobulinemia, peripheral neuropathy). Complications related to Sjogren's syndrome include for example infection of the parotid gland (typically staphylococcal, streptococcal, or pneumococcal) emergence of parotid tumors, risk for fetal loss in pregnant patients, emergence of pseudolymphomas and non- Hodgkin B-cell lymphomas. The gut microbiota has in the last few decades become a field of extensive research, and increasing evidence demonstrates the extent to which the bacteria colonizing the gut contribute to the development of immune mediated and metabolic disorders (Arora and Backhed, 2016). Identifying means to alleviate gut dysbiosis is much needed to help prevent, treat, and understand a number of such diseases. In this context, several approaches have been studied with a goal of promoting a healthy and well-balanced microbiota. Among these are pre- and probiotics with the potential of promoting growth of beneficial bacteria. US2016199424 discloses therapeutic compositions containing e.g. prebiotics, e.g., non- digestible medium and long-chained carbohydrates, in conjunction with microbial populations and/or networks thereof for use in method of reducing (treating) autoimmune or inflammatory disorders. US2016199424 also discloses a large number of various probiotics and/or prebiotics which allegedly are suitable for treatment of a large number of diseases associated with gut dysbiosis. US2016324904 discloses e.g. a dietary supplement for improving e.g.

autoimmune diseases by ingestion of mixtures of at least one prebiotic, such as dietary fibers and oligosaccharides, and at least one of L-glutamine, zinc, or vitamin A. The supplement is for improving immunotolerance, intestinal environment and intestinal tract barrier. The probiotic of US2016324904 may be a spore lactic acid bacteria, and the prebiotic may be an indigestible food ingredients such as oligosaccharide,

dietary fiber (polysaccharides), substance stimulating production of Lactobacillus bifidus and cyclic oligosaccharide.

WO14150566 relates to a method for reducing autoimmune disease using peptides selected from a casein hydrolysate. In a preferred embodiment of WO14150566 the nutritional composition comprises about 0.1 to about 1 g/100 kcal of prebiotic composition, wherein the prebiotic composition comprises at least 20 percent of an oligosaccharide.

US2007134391 discloses functional ingredient comprising a prebiotic, such as

dietary fibers, including, xylooligosaccharides, for the treatment and/or prevention of autoimmune disorders.

Dwivedi et al. 2016 discloses the importance of T regulatory cells (Tregs) in e.g. treatment of autoimmunity. Dwivedi et al. 2016 also discloses the impact of probiotics and prebiotics and the associated metabolites (short chain fatty acids, preferably butyrate) on the differentiation and function of Tregs. Dwivedi et al. 2016 also mentions that numerous autoimmune diseases have been shown to present significant number depletion and/or function impairment of Tregs. Hansen et al. 2016 discloses inter alia that germ-free (GF) mice had a very low degree of sialitis, which was restored by colonization with select microbial lineages. Moreover, Hansen et al. 2016 concludes that the severity of sialitis in the tested mice did not correlate with the degree of insulitis in the same animal, i.e. there was a distinction in relation to the requirements for the microbiota for different autoimmune

pathologies within the same organism.

Currently, neither a cure for Sjogren's syndrome nor a specific treatment is known which is capable of permanently restoring gland secretion. Instead, treatment is generally symptomatic and supportive. Moreover, no prophylactic treatment mechanism for Sjogren's syndrome is currently available due to its complexity as an autoimmune disorder. However, lifestyle changes can reduce the risk factors of getting Sjogren's syndrome or reduce the severity of the condition with patients who have already been diagnosed. Likewise, diet is strongly associated with inflammation that is mostly seen in many autoimmune related diseases including Sjogren's syndrome.

It is therefore one of the objects of the present invention to provide compounds, compositions and formulations for use in treatment of Sjogren's syndrome.

Summary of the invention

Using diets to fuel advantageous bacteria in the gut have proven beneficial for alleviating inflammation and disease driven by unfavorable gut microbiota composition (dysbiosis). Prebiotic compounds or compositions, such as xylooligossacharides (XOS) containing compounds or compositions are non-digestible and fermentable fiber compounds and source of short-chain fatty acids that the host digesting it can utilize.

The inventors have surprisingly discovered that XOS-supplemented diets besides other anti-inflammatory effects also have the ability to reduce cellular infiltrations in the salivary glands (sialitis) in non-obese diabetic (NOD) mice prone to the development of Sjogren's syndrome.

The inventors have now demonstrated that dietary XOS have the ability to reduce the inflammation caused by autoimmune cellular reactions against salivary glands in NOD mice.

Moreover, the inventors have established that microbes may not be necessary for XOS to mediate protection which is a surprising observation compared to the traditional perception of how prebiotics are typically perceived as a source of energy only by the symbiotic elements of the gut microbiota.

To understand the involvement of the intestinal milieu in complex autoimmune diseases, one has to consider both the signals balanced by symbionts and pathobiont but also those deriving directly from dietary antigens.

Eating prebiotic supplement is a more feasible approach that due to its advantage on mucus production may be able to limit environmental triggers constantly in risk of crossing the otherwise compromised barrier of the gut in preclinical individuals predisposed of autoimmune diseases.

In particular, it is an object of the present invention to provide a therapy that solves the above mentioned problems of the prior art with being unable to satisfactorily reduce salivary gland inflammation in patients suffering from Sjogren's syndrome.

Thus, one aspect of the invention relates to an orally administered prebiotic compound or composition which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in treatment of Sjogren's syndrome in the mammal.

Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in treatment of Sjogren's syndrome in the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in treatment of Sjogren's syndrome in the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in treatment of Sjogren's syndrome in the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in treatment of 5 Sjogren's syndrome in the mammal.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in treatment of Sjogren's syndrome in a mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5- 10 30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in treatment of Sjogren's syndrome in a mammal, where the XOS are administered 15 1-3 times daily for a total period of at least 7 days in amounts equal to 1-50%, such as 2- 45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

20 Another aspect of the invention relates to an orally administered prebiotic compound or composition which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in treatment of Sjogren's syndrome in the human subject.

25

Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a human subject and 30 (ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in treatment of Sjogren's syndrome in the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

35 (i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in treatment of Sjogren's syndrome in the human subject. Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, 5 where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-

40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in treatment of Sjogren's syndrome in the human subject.

10 Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-

15 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in treatment of Sjogren's syndrome in the human subject.

20 In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in treatment of Sjogren's syndrome in a human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

25

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in treatment of Sjogren's syndrome in a human subject, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1- 50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, 30 such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily

carbohydrate fraction ingested.

Another aspect of the invention relates to an orally administered prebiotic compound or composition which :

35 (i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal. Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten

monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested. In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in reducing cellular infiltrations in the salivary glands (sialitis) of the mammal, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, 5 such as 6-25%, such as 7-20%, such as 8- 15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

Another aspect of the invention relates to an orally administered prebiotic compound or composition which :

10 (i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human subject.

15 Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten

monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, 20 for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human

subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

25 (i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human subject.

30 Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-

35 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human subject. Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-

40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human subject.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in reducing cellular infiltrations in the salivary glands (sialitis) in the human subject, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

Another aspect of the invention relates to an orally administered prebiotic compound or composition which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in increasing saliva production in the oral cavity of the mammal. Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten

monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in increasing saliva production in the oral cavity of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and (ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in increasing saliva production in the oral cavity of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in increasing saliva production in the oral cavity of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in increasing saliva production in the oral cavity of the mammal.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing saliva production in the oral cavity of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4- 35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing saliva production in the oral cavity of the mammal, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1- 50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily

carbohydrate fraction ingested.

Another aspect of the invention relates to an orally administered prebiotic compound or composition which :

(i) remains essentially non-digested until it reaches the colon in a human subject and (ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in increasing saliva production in the oral cavity of the human subject.

Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in increasing saliva production in the oral cavity of the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in increasing saliva production in the oral cavity of the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and (ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in increasing saliva production in the oral cavity of the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-

40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in increasing saliva production in the oral cavity of the human subject.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing saliva production in the oral cavity of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) 5 for use in increasing saliva production in the oral cavity of the human subject, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6- 25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

10

Another aspect of the invention relates to an orally administered prebiotic compound or composition which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal,

15 for use in increasing tear production of the mammal.

Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten monosaccharide units which :

20 (i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in increasing tear production of the mammal.

Still another aspect of the invention relates to an orally administered prebiotic compound 25 or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, for use in increasing tear production of the mammal.

30 Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-

35 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in increasing tear production of the mammal. Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal,

5 where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in increasing tear production of the mammal.

10

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing tear production of the mammal, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8- 15% most preferably approx. 5% of the total 15 daily carbohydrate fraction ingested.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing tear production of the mammal, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1-50%, such as 2- 20 45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

Another aspect of the invention relates to an orally administered prebiotic compound or 25 composition which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in increasing tear production of the human subject.

30 Another aspect of the invention relates to an orally administered prebiotic compound or composition selected from the group of oligosaccharides containing three to ten

monosaccharide units which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, 35 for use in increasing tear production of the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and (ii) can be fermented by the commensal gut bacteria in the colon of the human subject, for use in increasing tear production of the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and

(ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested, for use in increasing tear production of the human subject.

Still another aspect of the invention relates to an orally administered prebiotic compound or composition comprising xylooligosaccharides (XOS) which :

(i) remains essentially non-digested until it reaches the colon in a human subject and (ii) can be fermented by the commensal gut bacteria in the colon of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3- 40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5%, of the total daily carbohydrate fraction ingested,

administered 1-3 times daily for a total period of at least 7 days for use in increasing tear production of the human subject.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing tear production of the human subject, where the XOS are administered in amounts equal to 1-50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily carbohydrate fraction ingested.

In a still further embodiment, the present invention relates to xylooligosaccharides (XOS) for use in increasing tear production of the human subject, where the XOS are administered 1-3 times daily for a total period of at least 7 days in amounts equal to 1- 50%, such as 2-45%, such as 3-40%, such as 4-35%, such as 5-30%, such as 6-25%, such as 7-20%, such as 8-15% most preferably approx. 5% of the total daily

carbohydrate fraction ingested.

In a further aspect of the invention, the orally administered prebiotic compound or composition for any of the above described uses may be incorporated into or part of a drink, beverage, snack bar, tablets, lozenge, capsules, soft gels, gel caps, powders, liquids, functional food products, conventional food products, nutraceuticals, natural health products, nutritional compositions, infant formulas, dairy products, chewing gums, candies.

In an even further aspect of the invention, the orally administered prebiotic compound or composition for any of the above described uses may be administered prior to,

concomitantly with or following an oral administration of one or more probiotic(s), preferably bifidobacteria and/or lactobacilli containing probiotic(s) .

In a still further embodiment, the present invention relates to a method of treating

Sjogren's syndrome in a mammal, comprising oral administration into the mammal of a compound and/or composition which :

(i) remains essentially non-digested until it reaches the colon in a mammal and

(ii) can be fermented by the commensal gut bacteria in the colon of the mammal. Brief description of the figures

Figure 1 shows the effects on the gut microbiota . Principal coordinates analysis (PCoA) plot of 16S rRNA gene tag-encoded amplicon sequencing based on the un-weighted (qualitative, based on presence of bacteria strains) and weighted (quantitative, based on proportions of bacteria strains) UniFrac distance matrix as indicated . The plots illustrates feces samples from four weeks old female NOD mice on a standard chow diet (C, black, n=9) and XOS supplemented diet (X, grey, n= 10) . The plots show that XOS supplemented diet alters the gut microbiota profile of the mice (being dominated by parabacteroides and lachnospiraceae with anti-inflammatory properties and lack of Prevotella, Rikenellaceae and Bacteroides) which therefore clusters separately on the PCoA plots.

Figure 2 shows that XOS reduce autoimmunity. Histopathology score of salivary glands collected from 13 weeks old female NOD mice on either standard chow diet (C, n= 10) or XOS supplemented diet their whole life (X, n=9) . Lymphocytic inflammatory foci in salivary glands were scored in a blinded fashion on H&E stained 5 pm sections of wax embedded salivary glands from 13 weeks old mice. Eight sequential sections of each gland with 40 pm intervals were scored by counting the number of focal infiltrates and measuring the mean size of all foci per section. Foci size was measured with a Leica graticule with 10 xlO mm grid and the sialitis score was calculated as: Score = Nf x sf [mm ² ] where Tvf is the mean number of foci per section and sf is the average size of the foci per section. The plot shows that dietary XOS reduces number and size of inflammatory foci as collected into a combined 'sialitis score' in the salivary glands of NOD mice. Error bars represent SEM . **** indicate P <0.001. Figure 3 shows that XOS fermentation increases acetic acid contents. Gas chromatographic analysis of the short-chain fatty acids (SCFA) Acetic Acid and other SCFAs such as n-Butyrate Acid and Propionic Acid in colon content collected at 13 weeks of age in female NOD mice fed either a control diet (C, n= 13) or a XOS supplemented diet (X, n= l l). SCFA concentrations indicate bacterial fermentation and metabolic activity of the gut microbiota and the plotshows that dietary XOS increases the production of the SCFA acetic acid in the gut. Other SCFAs were not regulated by XOS. Error bars represent SEM. ** indicate P>0.01

Figure 4 shows that XOS improves the gut barrier. Spectrophotometric analysis of plasma Fitc dextran level two hours post oral gavage at seven weeks of age (n=6/group). The plot shows that dietary XOS decreases the gut permeability monitored by systemic uptake of orally fed fluorescence-conjugated dextran (FITC-dextran). Relative gene expression of Mucl in colon biopsies sampled at 13 weeks of age is also shown. The female NOD mice were fed standard chow diet (C, n= 15) or XOS supplemented diet their whole life (X, n=9). Error bars represent SEM . *P < 0.05, ** P < 0.01.

Figure 5 shows that XOS reduces intestinal inflammation. Flow cytometric analysis of cells isolated from pancreatic lymph node (PLN), mesenteric lymph node (MLN), and spleen (SPL) in 13 weeks old female NOD mice (n=7/group). Percentages of CD8 ⁺ NKT cells and cytotoxic CD8 ⁺ T cells are shown. Mice were fed either standard chow diet (C, n= 15) or XOS supplemented diet their whole life (X, n=8). Error bars represent SEM. ** P < 0.01, *** P < 0.001, **** P < 0.0001. P values <0.1 are shown. The plots show that XOS reduces the fraction of inflammatory cells both locally in the gut and systemically. The present invention will now be described in more detail in the following.

Detailed description of the invention

Definitions

Prior to discussing the present invention in further details, the following terms and conventions will first be defined :

Administration

According to the present invention, administration refers to any oral administration form, such as ingestion, consumption, eating, drinking or otherwise which will be well-known for the skilled person. Commensal gut bacteria

According to the present invention, commensal gut bacteria (sometimes also referred to as gut microbiota, gut microflora or gastrointestinal microbiota) refers inter alia to gut bacteria deriving benefit to its host without harming or benefiting the host individual.

Dietary fiber

In the context of the present application, dietary fiber refers to the edible parts of plants or analogous carbohydrates that are resistant to digestion and absorption in the human small intestine, with complete or partial fermentation in the large intestine. Dietary fiber includes polysaccharides, oligosaccharides, lignin, and associated plant substances. Dietary fibers promote beneficial physiologic effects including laxation, and/or blood cholesterol attenuation, and/or blood glucose attenuation. Dietary fiber means carbohydrate polymers with≥10 monomeric units, which are not hydrolyzed by the endogenous enzymes in the small intestine of humans. Dietary fiber, such as pectin and xylan, are fermentable in the large intestine and forms an important source of short-chain fatty acids with a nutritional role for the epithelial cells lining the gut barrier. However, the sorts of fiber matter, as some polysaccharides such as pectin and xylan, are associated with e.g. higher levels of proinflammatory and stress-related transcripts in the colon and diabetogenic potential in NOD mice whereas

e.g. XOS shows immunomodulatory effects that seem to be related to immune

homeostasis and tolerance development. Dietary fibre is considered the portion of plant cells that are resistant to hydrolysis and human digestion. This allows the fiber to reach the colon undigested, allowing fermentation by the microbiota. Inulin is metabolised in the gut the same way as dietary fibre. Since they are not absorbed in the small intestine, calorie value is significantly lower compared to carbohydrates. As a result of the dietary fibre reaching the colon, there is an increase in biomass. If appropriate liquid intake is met to stay hydrated, the fibres are able to ferment. This results in increased fecal bulk and frequency, promoting regularity and alleviating constipation. Functional food

In the context of the present application, a functional food refers to food similar in appearance to, or may be, a conventional food that is consumed as part of a usual diet, and is demonstrated to have physiological benefits and/or reduce the risk of chronic disease beyond basic nutritional functions, i.e. they contain bioactive compound. Examples of functional foods may be pro- or prebiotics, pro- or prebiotics used in infant formulas, folic acid fortified bread, sports bars (incl. vitamins, proteins, pro- or prebiotics etc.). NOD mice

Non-obese diabetic (NOD) mice have traditionally been used as an animal model for type 1 diabetes (being an autoimmune disease). However, as NOD mice spontaneously develops polyglandular autoimmune inflammation affecting organs like thyroid and salivary glands in a fashion very comparable to human Sjogren's syndrome they have also turned out to be a useful animal model for assessing Sjogren's syndrome. As one of the most prevalent multiorgan autoimmune diseases, Sjogren's syndrome is characterized (among other things) by dry mouth (xerostomia and salivary gland hypofunction) and dry eyes

(keratoconjunctivitis sicca). NOD mice similarly manifest several features of Sjogren's syndrome (lymphocytic infiltrations and progressive destruction of salivary and lachrymal glands) making it a suitable animal model for Sjogren's syndrome.

Inulin

In the context of the present application, inulin refers to a soluble dietary fiber. It is a naturally occurring oligosaccharide belonging to a group of carbohydrates known as fructans. Unlike most carbohydrates, inulin is non-digestible. This allows it to pass through the small intestine and ferment in the large intestine. Through the fermentation process, the inulin becomes healthy intestinal microbiota.. Nutraceuticals

I the context of the present application, nutraceuticals are concentrated forms of food or food constituents that can be taken in pills, powder, or other medicinal forms that have specific health benefits. Prebiotics

I the context of the present application, prebiotics are defined as a selectively fermented ingredient that allows specific changes, both in the composition and/or activity in the gastrointestinal microbiota that confer benefits upon host's well-being and health.

Prebiotics resist digestion in the small intestine, are fermented in the large intestine, contain less energy in the form of calories than starch and sucrose, and increase stool weight and frequency. However, not all dietary fibres are prebiotic. Prebiotics are a non- digestible food ingredient that improves human health by its selective fermentation by one or a limited number of beneficial bacteria entirely by the colonic flora, and stimulating their growth and/or activity. Dietary fibers can be classified into soluble and non-soluble fibers. Soluble dietary fibers are fermented, but usually not in a selective way. Prebiotics fibers are fermented in a selective way, meaning not all soluble dietary fibers are prebiotic. Non- soluble dietary fibers are not fully fermented by the colonic flora. Prebiotics are fully fermented by the colonic microorganisms. Since some non-soluble dietary fibers are fermented outside the colon, not all non-soluble dietary fibers are prebiotic. Therefore, not all dietary fibers are prebiotic.

Thus, the most common type of prebiotic is from the soluble dietary fiber inulin. Inulin is common in many plants containing fructan. Furthermore, many of these plants are frequently eaten as vegetables - asparagus, garlic, leek, onion, and artichoke are excellent sources of inulin.

Probiotics

In the context of the present application, probiotics covers a group of diverse bacteria species, including bifidobacteria and lactobacilli, being known probiotics in humans, which are able to ferment e.g. XOS and can, thus, specifically be targeted to propagate in a host that ingests a prebiotic supplemented diet. Probiotics are living microorganisms that individually may provide the host a direct health benefit whereas prebiotics are defined as a selectively fermented ingredient that allows specific changes, both in the composition and/or activity in the gastrointestinal microbiota that confer benefits upon host's well- being and health. Xylooligossacharides (XOS) are interesting in this regard, as they elicit many of these desired properties. Probiotics are able to ferment e.g. XOS can specifically be targeted to propagate in a host that ingests a prebiotic supplemented diet. This may be beneficial for inflammatory autoimmune diseases regulated by the gut microbiota in humans in a similar manner as in the murine models. In the NOD mouse the inventors have found that XOS further improves the gut barrier and induces less proinflammatory cells in the gut. Sia litis

In the context of the present application, sialitis refers to inflammation of a salivary gland tissue.

Sjogren's syndrome

Sjogren's syndrome (sometimes also referred to as Sjogren's disease or disorder) is a systemic chronic inflammatory disorder characterized by lymphocytic infiltrates in exocrine glands. Most individuals with Sjogren's syndrome present with sicca symptoms, such as dry eyes, dry mouth (xerostomia), and parotid gland enlargement. Primary Sjogren's syndrome occurs in the absence of another underlying rheumatic disorder, whereas secondary Sjogren's syndrome is associated with another underlying rheumatic disease, such as systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), or scleroderma. In the present application, any reference to "Sjogren's syndrome" covers both primary and secondary Sjogren's syndrome. Sjogren's syndrome symptoms

Most Sjogren's syndrome patients (approx. 90%) are women, and onset is usually at age 40-60 years, but the syndrome also can affect men and children. The most prominent clinical disease manifestations include hyposalivation and keratoconjunctivitis sicca resulting in symptoms of oral and ocular dryness. . In children, bilateral parotid swelling is the most common sign of onset. Symptoms of Sjogren's syndrome can severely decrease the patient's quality of life in terms of its physical, psychological, and social aspects.

Patients may experience the effects xerostomia (dry mouth)and hyposalivation in the following ways: inability to eat dry food items because it sticks to the roof the mouth, tongue sticking to the roof of the mouth, difficulty speaking for long periods of time or the development of hoarseness, higher incidence of dental caries and gingival inflammation, altered sense of taste, development of oral candidiasis with angular cheilitis, which can cause pain in the oral mucosa. Also the ocular dryness and decreased tears production cause symptoms of itching and burning as well as sand- and gravel sensation in the eyes, and also increase the risk of ocular infections.

Treatment (of Sjogren's syndrome)

In the context of the present application, "treatment" of Sjogren's syndrome involves curative treatment as well as treatment which alleviates, lessens or removes Sjogren's syndrome manifestations/symptoms.

Xerostomia

In the context of the present application, xerostomia refers to is dryness in the mouth which may be associated with a change in the composition of saliva, or reduced salivary flow (hyposalivation), or have no identifiable cause. Thus, xerostomia is the subjective feeling of oral dryness, which is often (but not always) associated with hypofunction of the salivary glands. Xerostomia may be caused by autoimmune damage to the salivary glands. Sjogren's syndrome is one such disease. Xylooligossacharides (XOS)

In the context of the present application, xylooligossacharides (XOS) refers to XOS consisting of monomers of xylose units linked by β-1,4 glycosidic bonds, found in various food sources, and can be chemically or enzymatically produced from lignocellulosic containing sources such as corn cobs. XOS passes undigested through the gastrointestinal tract until it reaches the ascending colon where the saccharide molecules are fermented by residing gut bacteria. A diverse range of bifidobacteria and lactobacilli are able to ferment XOS and can, thus, specifically be targeted to propagate in a host that ingests a prebiotic supplemented diet. It should be noted that embodiments and features described in the context of one of the aspects of the present invention also apply to the other aspects of the invention.

All patent and non-patent references cited in the present application, are hereby incorporated by reference in their entirety.

The invention will now be described in further details in the following non-limiting examples. Examples

Example 1 (NOD mice experiments)

Reducing various disease symptoms by improving gut dysbiosis using dietary formulations is well-known.

One of the challenges, however, is to investigate whether an effect shown in a mouse model is also observed in human patients. In this case disease pathogenesis seems to be quite similar in the mouse compared to humans, but as the effect is elicited partially through the gut microbiota the qualitative and quantitative differences in gut microbiota composition between the two species is essential. In case of an effect in humans, it will additionally be important to get a picture of how far in the disease progress the subjects (patients) can be and still gain beneficial effect of e.g. XOS intake. This has been investigated in the NOD mouse model, where 12 weeks old adult mice with full degree of sialitis are given XOS diet to test the late stage effect. Also, doses for humans need to be validated in humans.

The inventors have previously tested (unpublished) whether a xylooligosaccharide (XOS) feed supplement fed whole life was capable of reducing sialitis in NOD mice. NOD mice are prone to develop lymphocytic infiltrations in several organs, including the salivary glands. Histological analysis of these was, thus, made to assess whether the XOS supplemented diet would reduce the degree of sialitis. The effect on foci size and numbers in the salivary gland was evidently very pronounced in 13 weeks old mice. NOD mice - experimental set up

Dams of the strain NOD/MrkTac from Taconic (Germantown, NY) were housed at the barrier protected animal facility, Faculty of Health and Medical Sciences, University of Copenhagen under standard conditions in open cages without filter lids. Housing and health monitoring was conducted according to FELASA guidelines.

Mothers and female offspring were fed ad libitum a modified 1430 Altromin natural diet (Lage, Germany) supplemented with either XOS (Shandong Longlive, Qingdao, China) or standard corn starch as control to balance the level of polysaccharides in the diets in amounts equal to 10% of the carbohydrate fraction.

Effect of XOS-diet on gut microbiota

To verify the effects of XOS-diet on the gut microbiota, the fecal microbiota was sequenced by tag-encoded 16S rRNA gene targeted amplicon high-throughput sequencing. Principal coordinates analysis showed distinct clusters already at four weeks of age due to diet (ANOSIM based on unweighted and weighted UniFrac distance matrices: P < 0.05, R = 0.22 and P = 0.11, R = 0.10 respectively; Figure 1).

The rRNA sequences of the tag-encoded 16S rRNA gene were:

(341F )NXt_16s_F: 5'-

TCGTCGGCAGCGTCAGATGTGTATAAGAGACAGCCTAYGGGRBGCASCAG -3'

(806R: )NXt_16s_R: 5'- GTCTCGTGGGCTCGGAGATGTGTATAAGAGACAGGGACTACNNGGGTATCTAAT -3'

This separation was mainly driven by segmented filamentous bacteria and Parabacteriodes, with both showing a relatively higher abundance in the young XOS-fed mice. The alpha diversity was significantly reduced in XOS-fed mice, which in part was due to a bloom in Parabacteroides that constituted on average 60% of the adult microbiota in XOS-fed mice, whereas Rikenellaceae and Bacteroides were almost absent in XOS-fed mice compared to control-fed mice (not shown).

It has been shown before, that prebiotics can improve glucose homeostasis and insulin sensitivity by e.g. short chain fatty acid (SCFA)-mediated activation of e.g. intestinal gluconeogenesis. However, 10% XOS supplementation had no effect on weight gain, blood glucose levels, serum leptin and insulin, or insulin resistance even though fermentation of XOS did increase the production of acetic acid in the colon but not n-butyric acid and propionic acid which remained unaltered. The inventors therefore concluded that the anti- inflammatory effect of XOS on the host was not mediated through any major shifts in metabolism. Effect ofXOS-diet on gut morphology

The inventors also examined the effects of the XOS-diet on the intestinal barrier morphology as prebiotics have also previously shown to alter gut morphology and function. A FITC-dextran assay was used to evaluate the gut integrity and the barrier was indeed less permeable in mice fed the XOS supplemented diet compared to mice fed control diet (Figure 4). Gene expression of mucus related genes were up-regulated in mice on a XOS diet (Figure 4).

Anti-inflammatory effects of XOS-diet

Potentially anti-inflammatory effects of XOS diet was analyzed by flow cytometry both locally and systemically. CD8+ NKT cells and cytotoxic CD8+ T cells were lower in XOS-fed mice compared to control-fed mice, both in the pancreatic- and mesenteric lymph node as well as in the spleen (Figure 5). These cells are known to infiltrate the organs as the salivary glands in NOD mice.

These observations were confirmed by qPCR of Cd8a, Gzmb (Granzyme B), and Fast (Fas ligand) gene expression which was similarly down-regulated in XOS-fed mice. These are all genes related to CD8 T cell cytotoxicity. Additionally, there were no differences in other anti-inflammatory cell populations: FoxP3+ regulatory T cells and CD103+ T cells, except for a minor reduction in CD103+ FoxP3+ regulatory T cells in the spleen of XOS-fed mice (8.7% ± 0.2) compared to control-fed mice (8.2% ± 0.04) P < 0.05).

Example 2 (murine intervention study)

A murine intervention study with the aim to determine, whether XOS diet also has a protective effect later in life after histopathological changes occur. Methods

1) Murine intervention study:

8 weeks old NOD mice are randomly divided into two groups of 12 mice in each. One group is fed ad libitum a modified 1430 Altromin natural diet (Lage, Germany) supplemented with either XOS (Shandong Longlive, Qingdao, China) or standard cornstarch as control to balance the level of polysaccharides in the diets in amounts equal to 10% of the carbohydrate fraction. Lymphocytic inflammatory foci are scored in a blinded fashion on H&E stained 5 pm sections of wax embedded salivary glands from 20 weeks old non-diabetic NOD mice.

Salivary flow is tested with pilocarpine a week before euthanization.

Salivary cytokines are measured with a pro-inflammatory mesoscale kit.

Potential follow-up studies

NOD mice treated with antibiotics from 5-8 weeks of age are transplanted with murine or human microbiota from control and XOS dietary groups to investigate the sufficiency of the microbiota to transfer the protective phenotype.

The intervention study on mice according to Example 2 will be carried out in the priority year.

Example 3 (human intervention study)

The inventors believe that ingestion of XOS and/or other oligosaccharides fermented in the gut can alleviate disease also in patients with Sjogren's syndrome. A human intervention study will be performed to provide proof-of-principle (by Assoc. Prof., PhD, DDS Anne Marie Lynge Pedersen, Department of Odontology, SUND-UCPH).

Experimental protocol

2 x 30 patients (18-60 years of age), i.e. test and control group, fulfilling the American- European Consensus Classification Criteria (AECC) for the diagnosis of primary Sjogren's syndrome, are recruited among patients referred to the Clinic for Oral Medicine,

Department of Odontology, Faculty of Health and Medical Sciences, University of

Copenhagen. Fertile women included in the study must use adequate contraception.

Patients who are in systemic treatment with cytotoxic drugs, and patients with active infection with fevers requiring antibiotic treatment, cannot be included in the study.

Interviews will be made using of standardized questionnaire including questions about the general and oral health, including the incidence of diseases and various medical conditions, symptoms of mouth and eye dryness and symptoms from other body structures, past and current medications, tobacco and alcohol consumption, daily fluid intake, dietary habits and dental care habits. The patients will be dosed daily with XOS diluted in water in a dose equivalent to 1-10 % of daily carbohydrate intake.

The clinical study include the registration of dental status (Decayed Missed Filled teeth and surfaces), registration of periodontal conditions through plaque and gingival indices, pocket depth (PD) and attachment loss (CAL) at 6 sites per tooth. Oral mucosal changes will also be registered.

Sialometri will be performed measuring the unstimulated and chewing-stimulated total salivary secretion within 15 min. and 5 min, respectively. The collected saliva will be placed on ice, distributed in Eppendorf tubes and stored in a freezer until later miRNA, proteom- and microbiome analysis. Labial salivary gland biopsies will be carried out using local anesthesia, where the procurement of 6-8 small salivary glands will investigated for the degree of lymphocytic infiltration (focus score) as routine histological analysis. 1-2 salivary glands will be removed for miRNA and proteomics. Cytokines will be monitored in the blood, and EULAR Sjogren's syndrome disease activity index (ESSDAI) will be made. Test of tear secretion by means of Schirmer's test will be performed. miRNA analysis is performed on saliva, salivary gland tissue and serum: Total RNA purified after which a panel of 48 miRNA analyzed. Proteome analysis is performed using a 2- dimensional electrophoresis (2DE) and MALDI-TOF-MS, while Western blot analysis and ELISA is used to validate the results of 2DE analyzes. HOMINGS (Human Oral Microbiome) identification is made using Next Generation Sequencing analysis performed on whole saliva, where up to 600 different bacteria can be identified.

Reducing disease symptoms by improving dysbiosis using dietary formulations is a favorable approach. One of the challenges is to investigate how far in the disease progress the patients can be and still gain beneficial effect of XOS intake. This will be possible by including subjects with different stages of Sjogren's syndrome and test the effect of the XOS supplement on disease severity.

The human intervention study will be carried out in the priority year. References

Arora and Backhed, 2016; "The gut microbiota and metabolic disease: current

understanding and future perspectives" , J Intern Med. 2016; 280: 339-349.

5 Dwivedi et al. 2016; "Induction of regulatory T cells: A role for probiotics and prebiotics tosuppress autoimmunity", Autoimmunity Reviews. 2016 (15) : 379-392.

Hansen et al. 2016, "Cutting Edge: Commensal Microbiota Has Disparate Effects on Manifestations of Polyglandular Autoimmune Inflammation" J Immunol. 2016; 197: 701- 10 705. Prepublished online 20 June 2016

Pedersen AML, Nauntofte B. The salivary component of primary Sjogren's syndrome: diagnosis, clinical features and management. In: Columbus F (ed): Arthritis Research 2005. New York. Nova Science Publishers, Inc., Chapter V, p. 105-46.

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Asmussen K, Pedersen AML, Holm L. Sjogrens syndrom. I: Reumatologi (Podenphant J, Jacobsen S, Manniche C, Stengaard-Pedersen K, Tarp U. (eds), FADL's Forlag, Kobenhavn, 3.udg., 2012: 319-31

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