Means and methods for the early prognosis of an individual's risk of contracting an autoimmune disease such as multiple sclerosis

Abstract

The present invention relates to means and methods for the early prognosis of a subject's risk of contracting an autoimmune disease such as multiple sclerosis. In particular, the invention relates to peptides useful for prognosis of an autoimmune disease such as multiple sclerosis. The peptides of the present invention can induce specific antibodies that cross-react with central nervous system (CNS)-derived proteins, thereby contributing to the manifestation of an autoimmune disease such as multiple sclerosis. The peptides of the present invention are thus useful to detect presence of said antibodies that cross-react with central nervous system (CNS)-derived proteins thereby allowing a prognosis of the risk of a subject to contract an autoimmune disease such as multiple sclerosis.

Description

[0001] Means and methods for the early prognosis of an individual’s risk of contracting an autoimmune disease such as multiple sclerosis

[0002] The present invention relates to means and methods for the early prognosis of a subject’s risk of contracting an autoimmune disease such as multiple sclerosis. In particular, the invention relates to peptides useful for prognosis of an autoimmune disease such as multiple sclerosis. The peptides of the present invention can induce specific antibodies that cross-react with central nervous system (CNS)-derived proteins, thereby contributing to the manifestation of an autoimmune disease such as multiple sclerosis. The peptides of the present invention are thus useful to detect presence of said antibodies that cross-react with central nervous system (CNS)-derived proteins thereby allowing a prognosis of the risk of a subject to contract an autoimmune disease such as multiple sclerosis.

[0003] Multiple sclerosis (MS) is a chronic autoinflammatory demyelinating disease of the CNS. A total of 2.8 million people are estimated to live with MS worldwide (35.9 per 100,000 population) h So far, there is no diagnostic test available that could predict the occurrence and onset of MS in an individual. Epstein-Barr Virus (EBV) is a ubiquitous herpesvirus that persistently infects more than 90% of the adult human population worldwide2. EBV is increasingly considered an important factor in MS pathogenesis3. There is a substantially higher risk of developing MS after primary EBV infection4, and this is especially linked to the development of high-level EBV EBNA-1- specific antibody titers in individual human hosts5. Mechanistically, this may be due to the molecular mimicry between a highly conserved region in the EBNA-1 region (EBNA381-452) that cross-react with distinct CNS-derived proteins and, thus, these antibodies have a potential role in molecular mimicry and autoreactivity6.

[0004] An early, even preclinical diagnosis of MS holds paramount importance in preventing further disability, as it facilitates effective disease management9. Initiating therapies as soon as possible in the disease course is crucial10,1h Research indicates that delaying treatment can negatively affect relapse rates, disability progression, and overall disease outcome12,13,14Furthermore, contemporary treatment approaches have shifted towards prioritizing early highly-effective treatment rather than treatment escalation15,16,17. This shift underscores the significance of early and proactive intervention for better disease outcomes.

[0005] While various prognostic markers have been identified, such as sex, certain functional system symptoms, and magnetic resonance imaging (MRI) findings, the most significant predictor of future disability is the early initiation of treatment18. This not only impacts patients directly but also has broader societal implications, as delayed treatment initiation is associated with productivity losses due to accumulation of disability19.

[0006] Currently, the primary treatment approach is to initiate therapy immediately after diagnosis. The repeatedly updated diagnostic criteria strongly underline the importance of earlier diagnosis so that treatment can be initiated sooner. There is some evidence that initiating treatment before the onset of symptoms, based on MRI findings suggestive of MS (radiologically isolated syndrome: RIS), can delay or potentially prevent the conversion to clinical definitive MS20. This suggests that there is a critical window of opportunity to identify patients highly susceptible to MS following EBV infection21.

[0007] Thus, there is an unmet need for the earliest timepoint to assess the risk of autoimmune diseases associated with EBV-specific antibodies that cross-react with CNS-derived proteins, such as MS, and to establish the diagnosis already in the preclinical, clinically asymptomatic phase.

[0008] The problem to be solved by the present invention is the provision of means and methods allowing early risk prognosis and / or early diagnosis of an autoimmune disease associated with EBV-specific antibodies that cross-react with CNS-derived proteins, such as MS.

[0009] The present invention solves the above stated problem. In particular, the above problem is solved by a peptide comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide. In particular, the invention is solved by a peptide comprising at least 16 to 72, preferably 16 to 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69. In particular, the invention is solved by a peptide selected from the group consisting of SEQ ID NOs: 1-53.

[0010] The inventors have found that EBNA38i-452-derived antibody titers are a reliable marker for the future development of MS. This is demonstrated in Example 1, which shows that the presence of antibodies against different EBNA381-452 derived peptide fragments and pools thereof significantly predict if a subject will contract MS in about 1 to 4 years, such as 1-2 years or 3-4 years, prior to the subject’s clinically and MRI-based MS diagnosis. This time frame in turn allows early diagnosis of MS and an early treatment, which, as described above, is of paramount importance for delaying / preventing MS symptoms, delaying / preventing further progression of MS, and, thus, delaying / preventing future disability of the affected subject. The peptide of the present invention is used to detect the presence of said antibodies. Thus, the above problem is solved by the provision of the peptide of the present invention since said peptide is used to detect the presence of the antibodies derived from EBNA381-452 or peptide fragments thereof, which in turn allow early prognosis if a subject will contract MS. In addition, Example 4 shows that EBNA38i-452-derived antibody titers, which can be detected via the peptide(s) of the present invention can be used as a reliable cutoff for a significantly increased risk of a subject to contract MS in the future. Furthermore, the presence and frequency of episodes of high levels of EBNA38i-452-derived antibody titers indicates the timeframe in which a subject will contract MS which in turn allows for a personalized treatment of a subject depending on their prognosis. Treatment may then be initiated shortly before MS is most likely to occur or clinical monitoring of the subject may increase towards the timepoint at which MS is most likely to occur in the subject. Furthermore, the prognosis of years between EBV seroconversion and disease onset allows for a delay of treatment which in turn reduces adverse effects that may occur during the course of a treatment. For example, a subject with a prognosis of about 3 to 6 years may commence to treatment only about 3 years after EBV seroconversion to minimize adverse effects during the time before disease onset.

[0011] Furthermore, the problem of the present invention is solved by the provision of a composition comprising at least two peptides each comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides. In particular, the problem of the present invention is solved by the provision of a composition comprising at least two peptides each comprising 16 to 72, preferably 16 to 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69. In particular, the problem of the present invention is solved by the provision of a composition comprising at least two peptides selected from the group of SEQ ID NOs: 1-53.

[0012] Although it is sufficient to use a single / individual peptide of the present invention as discussed above, the inventors have further found that combinations of EBNA381-452 derived peptide fragments are especially useful for predicting a subject’s risk of contracting MS in later life. Example 1 shows that using combinations of EBNA381-452 derived peptide fragments for detection of EBNA38i-452-derived antibodies can even further increase the significance and thus confidence of MS prediction in a subject and thus lead to a surprisingly improved prognosis of MS. This has been shown for example when using at least two EBNA381-452 derived peptide fragments. Thus, the above problem is solved by the provision of a composition of the present invention.

[0013] Furthermore, the problem of the present invention is solved by the provision of an antigen comprising or consisting of the peptide of the present invention, or comprising or consisting of the composition of the present invention.

[0014] As discussed above, the peptide and composition of the present invention are especially useful for detecting EBNA38i-452-derived antibodies thereby predicting a subject’s risk of contracting MS later in life (see Example 1). Accordingly, it is especially advantageous to use said peptide or composition as an antigen for detecting said EBNA38i-452-derived antibodies.

[0015] Furthermore, the problem of the present invention is solved by the provision of a method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising detecting and / or quantifying binding of antibodies to the peptide of the invention, or the composition of the invention, or the antigen of the invention in a biological sample of said subject.

[0016] Furthermore, the problem of the present invention is solved by the provision of a method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising:

[0017] (a) contacting a biological sample of said subject with the peptide of the invention, or the composition of the invention, or the antigen of the invention,

[0018] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen. Furthermore, the problem of the present invention is solved by the provision of an in vitro or ex vivo method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising:

[0019] (a) contacting a biological sample of said subject with the peptide of the invention, or the composition of the invention, or the antigen of the invention,

[0020] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen. Furthermore, the problem of the present invention is solved by the provision of a peptide of the present invention, or the composition of present invention, or the antigen of present invention, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably MS.

[0021] The inventors have shown that the presence of EBNA38i-452-derived antibodies significantly correlates with later development of MS (see Example 1). Accordingly, the above-mentioned methods and diagnostic use solve the above stated problem insofar they detect the presence of EBNA38i-452-derived antibodies thereby allowing early prognosis and / or diagnosis of the associated disease. In addition, the already above discussed prognosis of a subject to be at risk of contracting MS and to be at risk of contracting MS in a certain number of years allows for unprecedented treatment options depending on the subject’s prognosis. This is shown in Example 4 and discussed above.

[0022] Furthermore, the above stated problem is used by the provision of a peptide selected from the group consisting of SEQ ID NOs: 20-26, 32-40 and 52-53, or a variant of said peptide having at least about 60% sequence identity to the peptide; and / or a composition comprising or consisting of the peptides of SEQ ID NOs: 20-26 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 32-40 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising or consisting of the peptides of SEQ ID NOs: 20-26, 32-40 and 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides.

[0023] The inventors have found that some EBNA381-452 derived peptide fragments do not significantly indicate that the subject may contract MS. Accordingly the above-mentioned peptides are useful as a negative control when assessing a subject’s risk for contracting MS, e.g. for validating the results of EBNA381-452 derived peptide fragments that significantly indicate a subject’s risk of contracting MS.

[0024] Furthermore, the above stated problem is solved by the provision of a use of the peptides of the invention, or of the compositions of the invention, as an antigen, for the same reasons explained above in relation to the peptides, compositions, and antigen of the present invention.

[0025] Furthermore, the above stated problem is solved by the provision of a kit comprising the peptides of the invention, or the compositions of the invention, or the antigen of the invention, and means for detecting and / or quantifying antibodies binding to said peptides, compositions, or antigen, for the same reasons explained above in relation to the peptides, compositions, and antigen of the present invention.

[0026] Preferred embodiments of the present invention are defined in the dependent claims.

[0027] Although the above paragraph refers to the Examples of the invention, it is noted that the invention is not limited to the Examples, e.g. limited to MS. The examples have been performed with MS as an exemplary autoimmune disease for illustrating the successful prognosis. However, these results can be extrapolated to any autoimmune disease associated with EB V-specific antibodies that crossreact with CNS-derived proteins. This is because the presence of the EBNA381-452 derived antibodies will contribute to the development of any autoimmune disease caused, facilitated and / or affected by a disturbance of the targeted endogenous CNS-derived proteins. Accordingly, the present invention plausibly shows the prognosis of any autoimmune disease associated with EBV- specific antibodies that cross-react with CNS-derived proteins. Such diseases are for example MS, myasthenia gravis (MG), systemic lupus erythematosus (SLE) or autoimmune encephalitis.

[0028] The invention provides the following items:

[0029] 1. A peptide comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0030] 2. The peptide of item 1 comprising 16 to 72, preferably 16 to 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69.

[0031] 3. The peptide of item 1 comprising 5 to 38 consecutive amino acids of the sequence set forth in SEQ ID NO: 55, or comprising 5 to 24 consecutive amino acids of the sequence set forth in SEQ ID NO: 56, or comprising 5 to 30 consecutive amino acids of the sequence set forth in SEQ ID NO: 57, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0032] 4. The peptide of any one of items 1 to 3, wherein the consecutive amino acids are, comprise and / or form a functional epitope.

[0033] 5. The peptide of any one of items 1 to 4, wherein the peptide is capable of eliciting an autoimmune response, and / or wherein the peptide is capable of binding antibodies or being bound by antibodies that cross-react with central nervous system (CNS)-derived proteins.

[0034] 6. The peptide of any one of items 1 to 5, wherein the peptide is selected from the group consisting of SEQ ID NOs: 1-53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0035] 7. The peptide of any one of items 1 to 6, wherein the peptide is selected from the group consisting of SEQ ID NOs: 1-19, 27-31 and 41-51, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0036] 8. The peptide of any one of items 1 to 7, wherein the peptide is the peptide of SEQ ID NO: 9, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0037] 9. The peptide of any one of items 1 to 8, wherein the peptide is comprised in a solid phase, a semi-solid phase or liquid phase.

[0038] 10. The peptide of item 9, wherein the peptide is immobilized in or on a semi-solid or solid surface.

[0039] 11. The peptide of item 10, wherein the peptide is coated onto the surface of a membrane, microtiter plate, bead, gel, or column or is immobilized in a gel.

[0040] 12. Two or more peptides of any one of items 1 to 11, wherein the peptides are covalently or partially covalently conjugated to each other.

[0041] 13. A composition comprising at least two peptides each comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides.

[0042] 14. The composition of item 13 comprising at least two peptides each comprising 5 to 38 consecutive amino acids of the sequence set forth in SEQ ID NO: 55, and / or comprising 5 to 24 consecutive amino acids of the sequence set forth in SEQ ID NO: 56, and / or comprising 5 to 30 consecutive amino acids of the sequence set forth in SEQ ID NO: 57, or variants of said peptides having at least about 60% sequence identity to the peptides.

[0043] 15. The composition of item 13 or 14, wherein the consecutive amino acids are, comprise and / or form a functional epitope.

[0044] 16. The composition of any one of items 13 to 15, wherein the at least two peptides are capable of eliciting an autoimmune response, and / or wherein the at least two peptides are capable of binding antibodies or being bound by antibodies that cross-react with central nervous system (CNS)-derived proteins. The composition of any one of items 13 to 16 comprising at least two peptides selected from the group of SEQ ID NOs: 1-53, or variants of said at least two peptides having at least about 60% sequence identity to the peptides. The composition of any one of items 13 to 17, comprising at least two peptides selected from the group of SEQ ID NOs: 1-19, 27-31 and 41-51, or variants of said at least two peptides having at least about 60% sequence identity to the peptides. The composition of any one of items 13 to 18, comprising

[0045] (i) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0046] (ii) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0047] (iii) at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0048] (iv) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide. The composition of any one of items 13 to 19, comprising the peptides of SEQ ID NOs: 1- 19 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 27-31 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides. The composition of item 20, consisting of the peptides of SEQ ID NOs: 1-19 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 27-31 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier, preferably consisting of the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier. The composition of any one of items 13 to 20, comprising the peptides of SEQ ID NOs: 1- 53 or variants of said peptides having at least about 60% sequence identity to the peptides. The composition of item 22 consisting of the peptides of SEQ ID NOs: 1-53 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier. The composition of any one of items 13 to 18 comprising the peptides of:

[0049] (a) SEQ ID NOs: 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0050] (b) SEQ ID NOs: 8, 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0051] (c) SEQ ID NOs: 8-12 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0052] (d) SEQ ID NOs: 7-13 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0053] (e) SEQ ID NOs: 7-13, 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0054] (f) SEQ ID NOs: 7-14, 29, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0055] (g) SEQ ID NOs: 7-15, 29, 43, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0056] (h) SEQ ID NOs: 3, 7-15, 29, 43, 45, 47 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0057] (i) SEQ ID NOs: 3-4, 7-15, 29, 43, and 45-48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0058] (j) SEQ ID NOs: 3-16, 29, 43, and 45-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0059] (k) SEQ ID NOs: 3-17, 29, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0060] (l) SEQ ID NOs: 3-17, 28-30, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0061] (m) SEQ ID NOs: 3-17, 19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0062] (n) SEQ ID NOs: 2-19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0063] (o) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0064] (p) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0065] (q) SEQ ID NOs: 2-19, 27-31, and 42-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination of the above. The composition of item 24, consisting of the peptides of:

[0066] (a) SEQ ID NOs: 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0067] (b) SEQ ID NOs: 8, 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0068] (c) SEQ ID NOs: 8-12 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0069] (d) SEQ ID NOs: 7-13 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0070] (e) SEQ ID NOs: 7-13, 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0071] (f) SEQ ID NOs: 7-14, 29, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0072] (g) SEQ ID NOs: 7-15, 29, 43, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0073] (h) SEQ ID NOs: 3, 7-15, 29, 43, 45, 47 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0074] (i) SEQ ID NOs: 3-4, 7-15, 29, 43, and 45-48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0075] (j) SEQ ID NOs: 3-16, 29, 43, and 45-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0076] (k) SEQ ID NOs: 3-17, 29, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0077] (l) SEQ ID NOs: 3-17, 28-30, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0078] (m) SEQ ID NOs: 3-17, 19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0079] (n) SEQ ID NOs: 2-19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0080] (o) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0081] (p) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0082] (q) SEQ ID NOs: 2-19, 27-31, and 42-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination thereof and a suitable carrier. The composition of any one of items 13 to 18 comprising the peptides of:

[0083] (i) SEQ ID NOs: 3 and 14 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0084] (ii) SEQ ID NOs: 3 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0085] (iii) SEQ ID NOs: 3 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0086] (iv) SEQ ID NOs: 14 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0087] (v) SEQ ID NOs: 14 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0088] (vi) SEQ ID NOs: 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination of the above. The composition of item 26 consisting of the peptides of:

[0089] (i) SEQ ID NOs: 3 and 14 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0090] (ii) SEQ ID NOs: 3 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0091] (iii) SEQ ID NOs: 3 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0092] (iv) SEQ ID NOs: 14 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0093] (v) SEQ ID NOs: 14 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0094] (vi) SEQ ID NOs: 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination thereof and a suitable carrier.

[0095] 28. The composition of any one of items 13 to 27, wherein the peptides are comprised in a solid phase, a semi-solid phase or liquid phase.

[0096] 29. The composition of item 28, wherein the peptides are immobilized in or on a semi-solid or solid surface.

[0097] 30. The composition of item 29, wherein the peptides are coated onto the surface of a membrane, microtiter plate, bead, gel or column or are immobilized in a gel.

[0098] 31. The composition of any one of items 13 to 30, wherein the at least two peptides are covalently or partially covalently conjugated to each other.

[0099] 32. An antigen comprising or consisting of the peptide of any one of items 1 to 12, or comprising or consisting of the composition of any one of items 13 to 31.

[0100] 33. A method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably multiple sclerosis (MS), the method comprising detecting and / or quantifying binding of antibodies to the peptide of any one of items 1 to 12, or the composition of any one of items 13 to 31, or the antigen of item 32 in a biological sample of said subject.

[0101] 34. A method for early prognosis of a subj ect being at risk of contracting an autoimmune disease, preferably multiple sclerosis (MS), the method comprising:

[0102] (a) contacting a biological sample of said subject with the peptide of any one of items 1 to 12, or the composition of any one of items 13 to 31, or the antigen of item 32,

[0103] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0104] 35. An in vitro or ex vivo method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably multiple sclerosis (MS), the method comprising:

[0105] (a) contacting a biological sample of said subject with the peptide of any one of items 1 to 12, or the composition of any one of items 13 to 31, or the antigen of item 32,

[0106] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0107] 36. The method of any one of items 33 to 35, wherein the method is performed between about 10 years and about 1 month prior to a diagnosis or manifestation of the autoimmune disease in the subject, preferably, wherein the method is performed at least 4, 3, 2, or at least 1 year prior to a diagnosis or manifestation of the autoimmune disease in the subject.

[0108] 37. The method of any one of items 33 to 36, wherein the method is performed between about 10 years and about 1 month after Epstein-Barr virus (EBV) seroconversion in the subject, preferably, wherein the method is performed between about 1 month and about 48 months after Epstein-Barr virus (EBV) seroconversion in the subject.

[0109] 38. The method of any one of items 33 to 37, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about every 1-12 months, such as every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 month(s).

[0110] 39. The method of any one of items 33 to 38, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or about 4 times after EBV seroconversion in the subject.

[0111] 40. The method of any one of items 33 to 39, wherein the autoimmune disease is selected from multiple sclerosis (MS), myasthenia gravis (MG), systemic lupus erythematosus (SLE), or autoimmune encephalitis.

[0112] 41. The method of any one of items 33 to 40, wherein the subject is at risk of contracting the autoimmune disease if binding of antibodies to said peptide, composition or antigen is detected and / or quantified.

[0113] 42. The method of any one of items 33 to 41, wherein the method further comprises step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control.

[0114] 43. The method of item 42, wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more healthy subject (negative control), and / or wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more subjects that have contracted the autoimmune disease (positive control).

[0115] 44. The method of any one of items 33 to 43, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject significantly deviates from the negative control, and / or wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject does not significantly deviate from the positive control.

[0116] 45. The method of item 44, wherein the significant deviation from the negative control is an increase in the detected and / or quantified binding in said biological sample of said subject compared to the negative control.

[0117] 46. The method of any one of items 33 to 45, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of = 450 nm and = 620 nm.

[0118] 47. The method of any one of items 33 to 45, wherein the subject is not at risk or at low risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0119] 48. The method of any one of items 33 to 45, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 8 years after EBV seroconversion if

[0120] (i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or at least about 4 times after EBV seroconversion in the subject, and

[0121] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0122] 49. The method of any one of items 33 to 45, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion if

[0123] (i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 2 or about 3 times after EBV seroconversion in the subject, and

[0124] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0125] 50. The method of any one of items 33 to 45, wherein the subject is not at risk or at low risk of contracting the autoimmune disease in about 8 to about 13 years after EBV seroconversion if

[0126] (i) no binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified after EBV seroconversion in the subject, and / or (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of = 450 nm and = 620 nm.

[0127] 51. The method of any one of items 33 to 50, wherein the method further comprises advising pre-clinical monitoring to avoid symptomatic manifestation of an autoimmune disease, and / or to allow early diagnosis of the autoimmune disease, e.g. MS, and / or wherein the method further comprises advising early treatment, if the subject is at risk of contracting the autoimmune disease and / or wherein the method further comprises advising a delay of treatment of the autoimmune disease, if the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

[0128] 52. The method of item 51, wherein the pre-clinical monitoring comprises frequent medical examination, such as neurological and radiological monitoring of the subject, e.g. via magnetic resonance imaging (MRI) of the subject’s brain and spinal cord, lumbar puncture, optical coherence tomography (OCT) and / or electrophysiological examination.

[0129] 53. The method of any one of items 33, 34 or 36 to 52, wherein the method further comprises treatment of the autoimmune disease if the subject is at risk of contracting the autoimmune disease or, wherein the method further comprises delaying of treatment of the autoimmune disease, if the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

[0130] 54. The method of any one of items 33 to 53, wherein the binding of antibodies is detected and / or quantified using Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC), Flow Cytometry and / or Fluorescence-Activated Cell Sorting (FACS) and the like.

[0131] 55. The method of any one of items 33 to 54, wherein the biological sample is a blood sample or a cerebrospinal fluid (CSF) sample.

[0132] 56. The method of item 55, wherein the blood sample is a whole blood sample, a blood serum sample, or a blood plasma sample.

[0133] 57. The method of any one of items 33 to 56, wherein the biological sample or blood sample or cerebrospinal fluid (CSF) sample is a human biological sample or a human blood sample or a human cerebrospinal fluid (CSF) sample. 58. The peptide of any one of items 1 to 12, or the composition of any one of items 13 to 31, or the antigen of item 32, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably multiple sclerosis (MS).

[0134] 59. The peptide, composition or antigen for use of item 58, wherein the method comprises:

[0135] (a) contacting a biological sample of a subject with the peptide any one of items 1 to 12, or the composition of any one of items 13 to 31, or the antigen of item 32,

[0136] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0137] 60. The peptide, composition or antigen for use of item 58 or 59, wherein the method is performed between about 10 years and about 1 month prior to a diagnosis or manifestation of the autoimmune disease in the subject, preferably, wherein the method is performed at least 4, 3, 2, or at least 1 year prior to the manifestation of the autoimmune disease in the subject.

[0138] 61. The peptide, composition or antigen for use of any one of items 58 to 60, wherein the method is performed between about 10 years and about 1 month after Epstein-Barr virus (EBV) seroconversion in the subject, preferably, wherein the method is performed between about 1 month and about 48 months after Epstein-Barr virus (EBV) seroconversion in the subject.

[0139] 62. The peptide, composition or antigen for use of any one of items 58 to 61, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about every 1-12 months, such as every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 month(s).

[0140] 63. The peptide, composition or antigen for use of any one of items 58 to 62, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or about 4 times after EBV seroconversion in the subject.

[0141] 64. The peptide, composition or antigen for use of any one of items 58 to 63, wherein the autoimmune disease is selected from multiple sclerosis (MS), myasthenia gravis (MG), or systemic lupus erythematosus (SLE) or autoimmune encephalitis.

[0142] 65. The peptide, composition or antigen for use of any one of items 58 to 64, wherein the subject is at risk of contracting the autoimmune disease if binding of antibodies to said peptide, composition or antigen is detected and / or quantified. 66. The peptide, composition or antigen for use of any one of items 58 to 65, wherein the method further comprises step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control.

[0143] 67. The peptide, composition or antigen for use of item 66, wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more healthy subject (negative control), and / or wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more subjects that have contracted the autoimmune disease (positive control).

[0144] 68. The peptide, composition or antigen for use of any one of items 58 to 67, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject significantly deviates from the negative control, and / or wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject does not significantly deviate from the positive control.

[0145] 69. The peptide, composition or antigen for use of item 68, wherein the significant deviation from the negative control is an increase in the detected and / or quantified binding in said biological sample of said subject compared to the negative control.

[0146] 70. The peptide, composition or antigen for use of any one of items 58 to 69, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of = 450 nm and = 620 nm.

[0147] 71. The peptide, composition or antigen for use of any one of items 58 to 69, wherein the subject is not at risk or at low risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-hsi- 452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0148] 72. The peptide, composition or antigen for use of any one of items 58 to 69, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 8 years after EBV seroconversion if

[0149] (i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or at least about 4 times after EBV seroconversion in the subject, and (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of = 450 nm and = 620 nm.

[0150] 73. The peptide, composition or antigen for use of any one of items 58 to 69, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion if

[0151] (i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 2 or about 3 times after EBV seroconversion in the subject, and

[0152] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0153] 74. The peptide, composition or antigen for use of any one of items 58 to 69, wherein the subject is not at risk or at low risk of contracting the autoimmune disease in about 8 to about 13 years after EBV seroconversion if

[0154] (i) no binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified after EBV seroconversion in the subject, and / or

[0155] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0156] 75. The peptide, composition or antigen for use of any one of items 58 to 74, wherein the method further comprises advising pre-clinical monitoring to avoid symptomatic manifestation of an autoimmune disease, and / or to allow early diagnosis of the autoimmune disease, e.g. MS, and / or wherein the method further comprises advising early treatment, if the subject is at risk of contracting the autoimmune disease, and / or wherein the method further comprises advising a delay of treatment of the autoimmune disease, if the subject is diagnosed with a risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

[0157] 76. The peptide, composition or antigen for use of item 75, wherein the pre-clinical monitoring comprises frequent medical examination, such as neurological and radiological monitoring of the subject, e.g. via magnetic resonance imaging (MRI) of the subject’s spinal cord and / or brain, lumbar puncture, optical coherence tomography (OCT) and / or electrophysiological examination.

[0158] 77. The peptide, composition or antigen for use of any one of items 58 to 76, wherein the method further comprises treatment of the autoimmune disease if the subject is diagnosed with a risk of contracting the autoimmune disease or, wherein the method further comprises delaying treatment of the autoimmune disease, if the subject is diagnosed with a risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

[0159] 78. The peptide, composition or antigen for use of any one of items 58 to 77, wherein the binding of antibodies is detected and / or quantified using any one of Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC) or Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS) and the like.

[0160] 79. The peptide, composition or antigen for use of any one of items 59 to 78, wherein the biological sample is a blood sample or a cerebrospinal fluid (CSF) sample.

[0161] 80. The peptide, composition or antigen for use of item 79, wherein the blood sample is a whole blood sample, a blood serum sample or a blood plasma sample.

[0162] 81. The peptide, composition or antigen for use of any one of items 59 to 80, wherein the biological sample or blood sample or cerebrospinal fluid (CSF) sample is a human biological sample or a human blood sample or a human cerebrospinal fluid (CSF) sample.

[0163] 82. A peptide selected from the group consisting of SEQ ID NOs: 20-26, 32-40 and 52-53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0164] 83. A composition comprising or consisting of the peptides of SEQ ID NOs: 20-26 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 32-40 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising or consisting of the peptides of SEQ ID NOs: 20-26, 32-40 and 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides.

[0165] 84. Use of the peptide of any one of items 1 to 12, or of the composition of any one of items 13 to 31, or of the peptide of item 82, or of the composition of item 83, as an antigen.

[0166] 85. A kit comprising the peptide of any one of items 1 to 12, or the composition of any one of items 13 to 31, or the antigen of item 32, and / or the peptide of item 82, and / or the composition of item 83, and means for detecting and / or quantifying antibodies binding to said peptide, composition, or antigen. 86. The kit of item 85, further comprising an anti-human Immunoglobulin G (IgG) antibody.

[0167] 87. The kit of item 85 or 86, wherein the means for detecting and / or quantifying antibodies binding to said peptide, composition or antigen are means to perform an Enzyme-Linked Immunosorbent Assay (ELISA).

[0168] Multiple sclerosis

[0169] Multiple sclerosis (MS) is a chronic autoimmune disease that affects the CNS, which includes the brain and spinal cord. It is characterized by the immune system mistakenly attacking the protective sheath (myelin) that covers nerve fibers (axons), causing disruption of signaling and transduction pathways and tracts between the brain and functional effector systems of the body, which in turn cause neurological complaints and symptoms in an affected individual. Over time, the disease can cause accumulating and permanent damage of CNS cells and structures. The exact cause of MS is unknown, but it is believed to involve a combination of genetic susceptibility and environmental factors, such as EBV infection. Symptoms of MS can vary widely and may include sensory symptoms such as numbness and tingling, muscle weakness, gait and walking impairment, vision problems, neurogenic bladder and bowel dysfunction, fatigue and cognitive dysfunction.

[0170] There are several disease courses of MS. Relapsing-Remitting MS (RRMS) is the most common form, characterized by clearly defined attacks of new or worsening neurological symptoms (relapses) followed by periods of partial or complete recovery (remissions). During remission, symptoms may disappear completely, or some symptoms may persist and become permanent. In Secondary Progressive MS (SPMS), the disease progresses after an initial RRMS course more steadily over time, with or without relapses and remissions. Most people who are initially diagnosed with RRMS will transition to SPMS. Primary Progressive MS (PPMS) is characterized by a gradual worsening of neurological function from the onset of symptoms, without relapses or remissions. PPMS is less common than RRMS. The course of MS is unpredictable, and the symptoms and progression can vary significantly from one person to another.

[0171] Peptides

[0172] A peptide is a short chain of amino acids linked together by peptide bonds. A peptide bond is a covalent chemical bond formed between the carboxyl group of one amino acid and the amino group of another amino acid. Peptides are distinct from proteins, which are typically longer chains of amino acids. The distinction between a peptide and a protein is not strictly defined by a certain number of amino acids, but generally, peptides are shorter than proteins.

[0173] The peptides of the present invention may comprise one or more epitope space. Also in the context of the present invention peptides of 5-20 amino acids are to be applied, an epitope that may be comprised in a peptide of the present invention may be 5-8 amino acids. Accordingly, in the context of the invention also peptides of 5-8 amino acids may be used, e.g. a peptide comprising 5 to 8 consecutive amino acids of the sequence set forth in SEQ ID NO: 54. Accordingly and also scientifically founded a peptide of the present invention which is for example 5-20 amino acids may comprise one or more epitope space that can induce an antibody response. In Example 1, peptides with 20 amino acids have been used. The peptide length was chosen to be long enough to be able to map several epitopes. At the same time, a peptide comprising 20 consecutive amino acids can be preferred since it may be short enough to avoid undesired folding structures / secondary structures that could prevent antibody binding.

[0174] Although peptides of 20 amino acids were used in Example 1, the length of a peptide of the present invention is not particularly limited.

[0175] The present invention provides peptides that are useful for detecting EBNA381-452 derived antibodies. Accordingly, the invention provides a peptide comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptide. In general, the peptide of the present invention is derived from the EBNA381-452 peptide which is set forth in SEQ ID NO: 54 and / or SEQ ID NO: NCBI. The peptide of the present invention can comprise any number of consecutive amino acids of the EBNA381-452 peptide. Accordingly, the invention provides a peptide comprising 5 to 72 consecutive amino acids, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, or 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide. In a preferred embodiment, the invention provides a peptide comprising 5 to 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide. In a preferred embodiment, the invention provides a peptide comprising 5 to 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptide. In a preferred embodiment, the invention provides a peptide comprising 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptide. In a preferred embodiment, the invention provides a peptide comprising 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide. In Example 1, functionality of an individual peptide comprising 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54 have been shown to be effective for predicting a subject’s risk of contracting MS. Accordingly, the invention provides a peptide selected from the group consisting of SEQ ID NOs: 1-53, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide selected from the group consisting of SEQ ID NOs: 1-53, or a variant of said peptide having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptide.

[0176] The peptide of the present invention may also comprise all amino acids of the EBNA381-452 peptide set forth in SEQ ID NO: 54. Accordingly, the invention provides a peptide comprising the amino acid sequence set forth in SEQ ID NO: 54. In a preferred embodiment, the invention provides a peptide comprising 5 to 72 consecutive amino acid of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0177] The peptide of the present invention can also comprise any number of consecutive amino acids of the amino acid sequence set forth in SEQ ID NOs: 55, 56, and / or 57. As shown in Figure 1 of Example 1, peptide fragments of said SEQ ID NOs correspond to peptides that significantly predict MS in a subject with high confidence. Thus, peptides comprising consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56 and / or 57 may be preferred herein. The present invention provides a peptide comprising consecutive amino acids over the entire length of SEQ ID NOs: 55, 56 and / or 57, i.e. also a combination of said SEQ ID NOs, e.g. when the SEQ ID NOs are combined into a single sequence comprising 92 amino acids. Accordingly, the present invention provides a peptide comprising 5 to 92 consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or a variant of said peptide having at least about 60% sequence identity to the peptide. The peptide of the present invention can comprise any number of consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57. Accordingly, the present invention provides a peptide comprising 5 to 92 consecutive amino acids, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38,

[0178] 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64,

[0179] 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90,

[0180] 91, or 92 consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or a variant of said peptide having at least about 60% sequence identity to the peptide. The peptide of the present invention may also comprise the entire amino acid sequence set forth in SEQ ID NOs: 55, 56, and / or 57. Accordingly, the invention provides a peptide comprising or consisting of the amino acid sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0181] The invention also provides a peptide comprising 5 to 38, preferably 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 55, or comprising 5 to 24, preferably 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 56, or comprising 5 to 30, preferably 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 57, or a variant of said peptide having at least about 60% sequence identity to the peptide

[0182] In general, any peptide comprising a number of consecutive amino acids of the EBNA381-452 peptide is particularly useful in the sense of the present invention if said peptides is, comprises or forms a functional epitope. As used herein “functional epitope” refers to the specific part of the peptide that is recognized by the immune system, particularly epitopes of EBNA381-452 derived peptides provided herein can induce production of antibodies that cross-react with CNS-derived proteins, i.e. are, comprise and / or form functional epitopes. The functional epitope is the precise amino acid sequence and three-dimensional structure within the peptide that comes into direct contact with the EBNA381-452 derived antibodies that cross-react with CNS-derived proteins. Functional epitopes can either be continuous (linear), consisting of a sequence of amino acids in a polypeptide chain, or conformational (discontinuous), where the epitope is formed by amino acids that are brought together in the three-dimensional folded structure of the peptide but might be distant from each other in the linear sequence. In Example 1 it has been shown that the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 form are, comprise or form a functional epitope since they can detect the presence of EBNA381-452 derived antibodies that cross-react with central nervous system (CNS)-derived proteins thereby predicting a subject’s risk of contracting MS. Accordingly, any of the peptides of the present invention may comprise and / or form a functional epitope. In a preferred embodiment, any of the peptides of the present invention may comprise and / or form a functional epitope recognized by EBNA381-452 derived antibodies that cross-react with CNS- derived proteins.

[0183] As mentioned before, the EBNA381-452 peptide can lead to the production of EBNA381-452 derived antibodies that cross-react with CNS-derived proteins, e.g. in a human. Those antibodies, also referred herein as autoantibodies, cross-react with CNS-derived proteins thereby leading to the development or progression of an autoimmune disease such as MS. Accordingly, the peptides of the present invention may be defined to elicit an autoimmune response in a subject, e.g. a human. These autoimmune response in form of cross-reacting antibodies can then be detected using the peptides of the present invention as antigens thereby allowing the prognosis of a subject’s risk of contracting an autoimmune disease such as MS. Accordingly, in one embodiment of the invention any of the peptides provided herein may be capable of eliciting an autoimmune response and / or capable of binding antibodies or being bound by antibodies that cross-react with CNS-derived proteins.

[0184] As mentioned above, the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 have been shown in Example 1 to detect the presence of EBNA381-452 derived antibodies that cross-react with CNS- derived proteins thereby predicting a subject’s risk of contracting MS. Thus, those peptides are particularly preferred in the sense of the present invention. Accordingly, the invention provides a peptide selected from the group consisting of SEQ ID NOs: 1-19, 27-31 and 41-51, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide selected from the group consisting of SEQ ID NOs: 1-19, 27-31 and 41-51, or a variant of said peptide having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%,

[0185] 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%,

[0186] 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,

[0187] 98%, or 99% sequence identity to the peptide.

[0188] As shown in Example 1, individual peptides are particularly useful for predicting a subject’s risk of contracting MS. Accordingly the invention provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 1, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 2, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 3, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 4, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 5, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 6, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 7, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 8, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 9, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 10, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 11, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 12, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 13, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 14, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 15, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 16, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 17, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 18, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 19, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 20, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 21, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 22, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 23, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 24, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 25, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 26, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 27, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 28, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 29, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 30, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 31, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 32, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 33, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 34, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 35, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 36, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 37, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 38, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 39, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 40, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 41, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 42, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 43, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 44, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 45, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 46, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 47, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 48, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 49, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 50, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 51, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 52, or a variant of said peptide having at least about 60% sequence identity to the peptide. The invention also provides a peptide, wherein the peptide is the peptide of SEQ ID NO: 53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0189] As discussed herein above, a peptide comprising 20 amino acids may comprise one or more epitope spaces. Thus, the invention also provides peptides comprising about 5 to 19, such as about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, or about 19 consecutive amino acids of any one of the sequences set forth in SEQ ID NOs: 1-53. This can be advantageous to obtain a minimal peptide sequence comprising a functional epitope space.

[0190] Accordingly, the invention provides in one aspect a peptide comprising about 5 to 19, such as about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, or about 19 consecutive amino acids of any one of the sequences set forth in SEQ ID NOs: 1-53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0191] Although the invention is not limited to individual peptides, individual peptides can be particularly useful for prognosis of an autoimmune disease, such as MS. As shown in Example 1, Table 2.1 the significance (here represented as difference in means) of individual peptides to prognose MS can vary. This corresponds to a higher confidence of the peptide to prognose a subject’s risk of contracting MS, if corresponding antibodies to the peptide are present in the subject. As evident from Table 2.1 the peptide set forth in SEQ ID NO: 9 results in the highest significance to prognose MS compared to all other peptides, which indicates a high confidence when using SEQ ID NO: 9 e.g. for prognosis of MS. Accordingly, the invention provides in a preferred aspect a peptide wherein the peptide is the peptide of SEQ ID NO: 9. As evident from Table 2.1, also other peptides show a high confidence in prognosis of MS. Accordingly, the invention provides in one aspect a peptide, wherein the peptide is selected from any one of SEQ ID NOs: 9, 10, or 8. The invention also provides a peptide, wherein the peptide is selected from any one of SEQ ID NOs: 9, 10, 8, 12, 11, 13, 7, 29, 48, or 45. The invention also provides a peptide, wherein the peptide is selected from any one of SEQ ID NOs: 9, 10, 8, 12, 11, 13, 7, 29, 48, 45, 14, 15, 43, 47, 3, 46, 4,6, 49, 5, 16, 17, 44, 28, 30, 51, or 19.

[0192] The above-mentioned individual peptides are particularly useful for prognosis of an autoimmune disease, such as MS. Also, for these peptides it can be advantageous to obtain a minimal peptide sequence comprising a functional epitope space.

[0193] Accordingly, the invention provides in one aspect a peptide comprising about 5 to 19, such as about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, or about 19 consecutive amino acids of any one of the sequences set forth in SEQ ID NOs: 9, 10, 8, 12, 11, 13, 7, 29, 48, 45, 14, 15, 43, 47, 3, 46, 4,6, 49, 5, 16, 17, 44, 28, 30, 51, or 19, preferably SEQ ID NOs: 9, 10, 8, 12, 11, 13, 7, 29, 48, or 45, more preferably SEQ ID NOs: 9, 10, or 8, most preferably SEQ ID NO: 9, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0194] The peptides of the present invention may be comprised in any suitable carrier, e.g. they may be comprised in a solid phase, a semi-solid phase or a liquid phase. Examples of the peptide being comprised in a solid phase is that the peptides are immobilized, e.g. coated onto a solid surface, coated onto a membrane, a microtiter plate, beads or a column. In the sense of the present invention it is preferred, that the peptides are comprised in a solid phase, e.g. coated onto the surface of a microtiter plate. This has been demonstrated in Example 1 and allows for the detection of EBNA381-452 derived antibodies that cross-react with CNS-derived proteins, e.g. via an ELISA assay using the peptides of the present invention as an antigen. Examples of peptides comprised in a semi-solid phase are peptides that are comprised in a gel or have been immobilized on or in a gel. Examples of peptides comprised in a liquid phase are peptides that dissolved in an appropriate liquid solution. Peptides of the present invention may be comprised in such a liquid phase before being immobilized as described above, e.g. before being coated onto the solid surface of a microtiter plate. The peptides of the present invention may be immobilized by any means known in the art, such as chemical bonding or physical adsorption. Accordingly, they may be immobilized via covalent bonding, physical adsorption, affinity tags, biotin-streptavidin interaction, selfassembled monolayers (SAMs), photoactivatable linkers, silanization. Accordingly, in one embodiment any of the peptides provided herein may be comprised in a solid phase, a semi-solid phase or liquid phase. In another embodiment any of the peptides provided herein may be immobilized in or on a semi-solid or solid surface. In another embodiment, any of the peptides provided herein may be coated onto the surface of a membrane, microtiter plate, bead, gel, or column or is immobilized in a gel. In a preferred embodiment, any of the peptides provided herein may be coated onto the surface of a microtiter plate.

[0195] In general, any of the peptides provided herein can be covalently or partially covalently conjugated to each other, e.g. such as the functional epitope of both is combined into a single polypeptide. This can be realized via conjugating two or more peptides of the present invention via peptide bonds. Methods for conjugating two or more peptides are known in the art and include, inter alia, conjugation via chemical synthesis through native chemical ligation (NCL), peptide splicing through intein mediated ligation, click chemistry, disulfide bond formation, enzymatic ligation, maleimide-thiol reaction, strategic use of amino acid side chains. Accordingly, the invention provides two or more peptides provided herein, wherein the peptides are covalently or partially covalently conjugated to each other.

[0196] Individual peptides comprising consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69 have been shown to significantly predict the risk of a subject contracting MS 1-2 or 3-4 years before diagnosis with high confidence, see Table 2 and section “Prediction of MS" of Example 1.

[0197] The invention also provides a peptide selected from the group consisting of SEQ ID NOs: 20-26, 32-40 and 52-53, or a variant of said peptide having at least about 60% sequence identity to the peptide. When used in a pool of all peptides of SEQ ID NOs: 1-53 have been shown in Example 1 to successfully predict the risk of a subject of contracting MS 1, 2, 3, or 4 years before diagnosis. When used individually, SEQ ID NOs: 20-26, 32-40 and 52-53 do not significantly correlate with a risk of a subject for contracting MS (see Figure 1). Accordingly, those peptides when used individually are particularly advantageous as a negative control, although they still predict a subject’s risk of contracting an autoimmune disease, such as MS, when used in a pool.

[0198] Accordingly, the invention also provides a peptide selected from the group consisting of SEQ ID NOs: 20-26, 32-40 and 52-53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

[0199] The invention also provides a composition comprising or consisting of the peptides of SEQ ID NOs: 20-26 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 32-40 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising or consisting of the peptides of SEQ ID NOs: 20-26, 32-40 and 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides.

[0200] As used herein, the term “X% sequence identity” describes the degree of sequence similarity between two amino acid sequences by expressing the percentage of amino acids in a first sequence that are identical to the corresponding amino acids in a second sequence when aligned for maximum correspondence. It is determined by aligning the two sequences for optimal comparison, which may involve introducing gaps in either of the sequences to achieve the best alignment. The alignment can be performed using various sequence comparison algorithms or programs known in the art, such as BLASTp. The “% sequence identity” value is calculated by taking the number of identical amino acid positions divided by the total number of amino acid in the shorter of the sequences (or the defined segment of comparison) and multiplying by 100. It serves as a quantitative representation of the similarity between two amino acid sequences.

[0201] A peptide of the present invention can comprise any number of amino acid substitutions as long as it can still be used according to the invention, e.g. as long as it still is, comprises and / or forms a functional epitope as defined herein. Accordingly, the present invention also provides variants of a peptide of the present invention having at least about 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or at least about 99% sequence identity when compared to the peptide. The present invention also provides variants of a peptide of the present invention having at least about 60% to about 99% such as, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or about 99% sequence identity when compared to the peptide. Preferably, the invention provides variants of a peptide of the invention having more than about 90%, such as 91, 92, 93, 94, 95, 96, 97, 98, or 99%, or at least about 90%, such as 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, or about 90-99%, such as 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence identity when compared to the peptide.

[0202] As mentioned above, the peptides of the present invention may be comprised in any suitable carrier. As used herein a “suitable carrier” can be any carrier for storing peptides, e.g. for later use. A suitable carrier can be useful for maintaining the stability and activity of the peptides. A suitable carrier may be selected from buffers, such as phosphate buffered saline (PBS) and tris buffered saline (TBS), and / or stabilizing agents such as sodium azide and thimerosal. Furthermore, peptides can be lyophilized (freeze-dried) and stored as a dry powder with or without the addition of stabilizers like mannitol or trehalose before lyophilization. In addition, peptides may be stored in a way to preserve their stability and activity. Peptides may be stored at -20°C or -80°C to maintain stability over long periods. Peptides may be stored in dark or opaque containers to protect them from light, which can cause degradation. Peptides may be stored in airtight containers to prevent oxidation and moisture absorption. Compositions

[0203] The present invention also provides a composition comprising at least two peptides each comprising 5 to 72, preferably 5 to 20, more preferably 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides. The peptides comprised in the composition of the present invention can overlap in their amino acid sequence, e.g. SEQ ID NO: 1 shares, e.g. overlaps, 19 of 20 amino acids with SEQ ID NO: 2. This applies to all peptides provided in SEQ ID NOs: 1-53. This overlap eventually leads to an improved exposure of antigen / epitope space when the peptides are used in the compositions of the present invention and avoids the omission of peptide sequence, e.g. comprising an epitope space, that may be lost by fragmenting the EBNA381-452 into peptides without any overlap. Accordingly a peptide of the present invention may overlap or share about 1 to 19, such as about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, or about 19 amino acids with any other peptide of the present invention. The overlap of peptides of the present invention may reduce the chance that important epitope spaces may be interrupted, e.g. by an arbitrary decision on a sequence / sequence length. Thus, an overlap of peptide sequence may increase the chance that one or more epitopes are present in one peptide. Overlapping peptide sequence may further lead an advantage that the same potential epitope may be present several times in a single composition.

[0204] Accordingly, the invention provides a composition comprising at least two peptides each comprising 5 to 72, preferably 5 to 20, more preferably 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides, wherein the amino acid sequence of the at least two peptides partially overlaps.

[0205] The invention also provides a composition comprising at least two peptides each comprising 5 to 72, preferably 5 to 20, more preferably 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides, wherein the amino acid sequence of the at least two peptides partially overlaps or shares about 1 to 19, such as about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, or about 19 amino acids with any other peptide of the present invention.

[0206] In summary, without being bound by theory, an overlap of sequence between peptides can be advantageous to improve the chances that any potential epitope space is comprised in the peptides, e.g. comprised in the composition of the present invention, thereby avoiding losing any potential epitope space. This is also reflected in the surprising data, that using a composition of two or more peptides for example all peptides set forth in SEQ ID NOs: 1-53 leads to a highly significant prediction of MS in a subject (see non-limiting Example 1). As mentioned above, these peptides comprise 20 amino acids which overlap / share 19 amino acids, respectively.

[0207] The above applies mutatis mutandis to any peptide, composition and / or method / use of the present invention, e.g. as defined in the appended claims.

[0208] Such compositions are particularly useful for prognosis of a subject being at risk of contracting an autoimmune disease, such as MS. As shown in Example 1, compositions of at least two peptides result in a further increased significance of prediction of MS in a subject and thus can further increase confidence of early prognosis if a subject will contract MS (see Tables 3-5 and section “Prediction of MP' of Example 1). Accordingly, the invention provides a composition comprising at least two peptides each comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides. The invention also provides a composition comprising at least two peptides each comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptides. The invention also provides a composition comprising at least two peptides each comprising 5 to 72 consecutive amino acids, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16,

[0209] 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42,

[0210] 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68,

[0211] 69, 70, 71, or 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides. In a preferred embodiment, the invention provides a composition comprising at least two peptides each comprising 5 to 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides. In a preferred embodiment, the invention provides a composition comprising at least two peptides each comprising 5 to 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptides. In a preferred embodiment, the invention provides a composition comprising at least two peptides each comprising 20 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides.

[0212] As mentioned above, peptides comprising consecutive amino acids of the fragments of EBNA381- 452 set forth in SEQ ID Nos: 55, 56, and / or 57 may be particularly preferred (cf. Table 2). The present invention provides a composition comprising consecutive amino acids over the entire length of SEQ ID NOs: 55, 56 and / or 57, i.e. also a combination of said SEQ ID NOs, e.g. when the SEQ ID NOs are combined into a single sequence comprising 92 amino acids. Accordingly, the invention provides a composition comprising at least two peptides each comprising 5 to 92 consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or variants of said peptides having at least about 60% sequence identity to the peptides. The invention also provides a composition comprising at least two peptides each comprising 5 to 92 consecutive amino acids, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,

[0213] 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52,

[0214] 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,

[0215] 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, or 92 consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or variants of said peptides having at least about 60% sequence identity to the peptides. In a preferred embodiment, the invention provides a composition comprising at least two peptides each comprising 5 to 20 consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or variants of said peptides having at least about 60% sequence identity to the peptides. In a preferred embodiment, the invention provides a composition comprising at least two peptides each comprising 20 consecutive amino acids of the sequence set forth in SEQ ID NOs: 55, 56, and / or 57, or variants of said peptides having at least about 60% sequence identity to the peptides.

[0216] The invention also provides a composition comprising at least two peptides each comprising 5 to 38, preferably 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 55, and / or comprising 5 to 24, preferably 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 56, and / or comprising 5 to 30, preferably 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 57, or variants of said peptides having at least about 60% sequence identity to the peptides.

[0217] The invention also provides a composition comprising at least two peptides of the peptides that have been shown to be capable of prognosing a subject’s risk of contracting MS in Example 1. Accordingly, the invention provides a composition comprising at least two peptides selected from the group of SEQ ID NOs: 1-53, or variants of said at least two peptides having at least about 60% sequence identity to the peptides. The invention also provides a composition comprising at least two and at most 53 peptides selected from the group of SEQ ID NOs: 1-53, or variants of said at least two peptides having at least about 60% sequence identity to the peptides. As shown in Figure 1, there are three peaks that represent peptides significant for prognosis of MS. It is especially advantageous to combine individual peptides from said three peaks in the composition of the present invention to further improve confidence of a prognosis of MS. Accordingly, in a preferred embodiment, the invention provides a composition comprising at least two peptides selected from the group of SEQ ID NOs: 1-19, 27-31 and 41-51, or variants of said at least two peptides having at least about 60% sequence identity to the peptides. In a preferred embodiment, the invention provides a composition comprising at least two peptides selected from the group of SEQ ID NOs: 1-19, 27-31 and 41-51, or variants of said at least two peptides having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptides.

[0218] As shown in Example 1, specific combinations of peptides in the composition of the present invention are advantageous in prognosing the risk of a subject contracting an autoimmune disease such as MS. It is particularly advantageous to combine at least two peptides, wherein each peptide is selected from one of the EBNA381-452 peptide fragments that result in highly significant peptides corresponding to the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51, respectively (see Figure 1). Accordingly, the invention provides a composition comprising

[0219] (i) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0220] (ii) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0221] (iii) at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0222] (iv) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide.

[0223] The invention also provides a composition comprising

[0224] (i) at least one, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 peptides selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one, such as 1, 2, 3, 4 or 5 peptides selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0225] (ii) at least one, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 peptides selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 peptides selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0226] (iii) at least one, such as 1, 2, 3, 4 or 5 peptides selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 peptides selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or

[0227] (iv) at least one, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19 peptides selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one, such as 1, 2, 3, 4 or 5 peptides selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 peptides selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide.

[0228] The invention also provides a composition comprising the peptides of SEQ ID NOs: 1-19 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 27-31 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides.

[0229] The invention also provides a composition consisting of the peptides of SEQ ID NOs: 1-19 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 27-31 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier, preferably consisting of the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier.

[0230] The invention also provides a composition comprising the peptides of SEQ ID NOs: 1-53 or variants of said peptides having at least about 60% sequence identity to the peptides.

[0231] The invention also provides a composition consisting of the peptides of SEQ ID NOs: 1-53 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier.

[0232] The invention also provides a composition comprising the peptides of SEQ ID NOs: 1-53 or variants of said peptides having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptides.

[0233] The invention also provides a composition consisting of the peptides of SEQ ID NOs: 1-53 or variants of said peptides having at least about 60% to about 99%, such as 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to the peptides and a suitable carrier. In the sense of the present invention a suitable carrier may be any of the solid phase, semi-sold phase or liquid phase as described above in section “Peptides”. Accordingly, the features described in relation to the solid phase, semi-sold phase or liquid phase in the section “Peptides” above apply mutatis mutandis to the compositions of the present invention. In particular, the composition may further comprise or further consist of an appropriate liquid solution wherein the at least two peptides are dissolved and / or solid phase, e.g. a solid surface, whereon the at least two peptides are immobilized such as a microtiter plate.

[0234] Although it is sufficient to use individual peptides for prognosis of e.g. MS as shown in Example 1, Table 2 and 2.1. the significance of the MS prognosis and thereby the confidence can be further increased by using a combination of peptides to detect antibodies against. As shown in Example 1 Table 5 and 7, compositions comprising at least two peptides of the present invention result in unmatched confidence for prognosis of a subject’s risk of contracting an autoimmune disease such as MS. Already using a combination of only two peptides e.g. SEQ ID NO: 9 and 10 increases the significance of the prognosis and thereby the confidence of the same compared to using an individual peptide for prognosis, see Example 1, Tables 5-7. Thus, the compositions comprising at least two peptides provided herein are particularly advantageous since they allow a prognosis of MS with high significance and confidence. Accordingly, the invention provides a composition comprising the peptides of

[0235] (a) SEQ ID NOs: 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0236] (b) SEQ ID NOs: 8, 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0237] (c) SEQ ID NOs: 8-12 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0238] (d) SEQ ID NOs: 7-13 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0239] (e) SEQ ID NOs: 7-13, 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0240] (f) SEQ ID NOs: 7-14, 29, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0241] (g) SEQ ID NOs: 7-15, 29, 43, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0242] (h) SEQ ID NOs: 3, 7-15, 29, 43, 45, 47 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0243] (i) SEQ ID NOs: 3-4, 7-15, 29, 43, and 45-48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0244] (j) SEQ ID NOs: 3-16, 29, 43, and 45-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0245] (k) SEQ ID NOs: 3-17, 29, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0246] (l) SEQ ID NOs: 3-17, 28-30, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0247] (m) SEQ ID NOs: 3-17, 19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0248] (n) SEQ ID NOs: 2-19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0249] (o) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0250] (p) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0251] (q) SEQ ID NOs: 2-19, 27-31, and 42-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination of the above.

[0252] The invention also provides a composition consisting of the peptides of:

[0253] (a) SEQ ID NOs: 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0254] (b) SEQ ID NOs: 8, 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0255] (c) SEQ ID NOs: 8-12 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0256] (d) SEQ ID NOs: 7-13 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0257] (e) SEQ ID NOs: 7-13, 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0258] (f) SEQ ID NOs: 7-14, 29, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0259] (g) SEQ ID NOs: 7-15, 29, 43, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0260] (h) SEQ ID NOs: 3, 7-15, 29, 43, 45, 47 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0261] (i) SEQ ID NOs: 3-4, 7-15, 29, 43, and 45-48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0262] (j) SEQ ID NOs: 3-16, 29, 43, and 45-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0263] (k) SEQ ID NOs: 3-17, 29, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0264] (l) SEQ ID NOs: 3-17, 28-30, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or (m) SEQ ID NOs: 3-17, 19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0265] (n) SEQ ID NOs: 2-19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0266] (o) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0267] (p) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0268] (q) SEQ ID NOs: 2-19, 27-31, and 42-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination thereof and a suitable carrier.

[0269] The invention also provides comprising the peptides of:

[0270] (i) SEQ ID NOs: 3 and 14 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0271] (ii) SEQ ID NOs: 3 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0272] (iii) SEQ ID NOs: 3 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0273] (iv) SEQ ID NOs: 14 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0274] (v) SEQ ID NOs: 14 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0275] (vi) SEQ ID NOs: 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination of the above.

[0276] The invention also provides a composition consisting of the peptides of:

[0277] (i) SEQ ID NOs: 3 and 14 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0278] (ii) SEQ ID NOs: 3 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0279] (iii) SEQ ID NOs: 3 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0280] (iv) SEQ ID NOs: 14 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0281] (v) SEQ ID NOs: 14 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or

[0282] (vi) SEQ ID NOs: 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination thereof and a suitable carrier. As described above in relation to the section “Peptides”, the peptides comprised in the compositions of the present invention can be, comprise and / or form a functional epitope. Thus, the features described in relation to a functional epitope in the section “Peptides” above apply mutatis mutandis to the peptides comprised in the compositions of the present invention.

[0283] As described above in relation to the section “Peptides”, the peptides comprised in the compositions of the present invention can be capable of eliciting an autoimmune response, and / or can be capable of binding antibodies or being bound by antibodies that cross-react with CNS- derived proteins. Thus, the features described in relation to eliciting an autoimmune response and / r binding or being bound by said antibodies in the section “Peptides” above apply mutatis mutandis to the peptides comprised in the compositions of the present invention.

[0284] As described above in relation to conjugated peptides in the section “Peptides”, the peptides comprised in the composition of the present invention may also be conjugated to each other. Accordingly, the features described in relation to the conjugated peptides in the section “Peptides” above, apply mutatis mutandis to the peptides comprised in the compositions of the present invention.

[0285] The peptides comprised in the composition of the present invention can comprise any number of amino acid substitutions as long as they can still be used according to the invention, e.g. as long as they still are, comprise and / or form functional epitopes as defined herein. Accordingly, the present invention also provides variants of peptides comprised in the composition of the present invention having at least 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or at least 99% sequence identity when compared to the peptides comprised in the composition. The present invention also provides variants of peptides comprised in the composition of the present invention having at least about 60% to about 99% such as, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or about 99% sequence identity when compared to the peptides comprised in the composition. Preferably, the invention provides variants of peptides comprised in the composition of the invention having more than 90%, such as 91, 92, 93, 94, 95, 96, 97, 98, or 99%, or at least 90%, such as 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99%, or 90-99%, such as 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% sequence identity when compared to the peptides comprised in the composition.

[0286] As used herein “at least two” such as “at least two peptides” means two or more, “at least two” may thus correspond to any number of peptides equal or greater than two. When used in the context of the composition of the present invention “at least two” may preferably comprise at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 18, at least 19, at least 20, at least 21, at least 22, at least 23, at least 24, at least 25, at least 26, at least 27, at least 28, at least 29, at least 30, at least 31, at least 32, at least 33, at least 34, at least 35, at least 36, at least 37, at least 38, at least 39, at least 40, at least 41, at least 42, at least 43, at least 44, at least 45, at least 46, at least 47, at least 48, at least 49, at least 50, at least 51, at least 52, and / or at least 53, and so on.

[0287] As mentioned above, the composition of the present invention may further comprise a suitable carrier. As used herein a “suitable carrier” can be any carrier for storing peptides, e.g. for later use. A suitable carrier can be useful for maintaining the stability and activity of the peptides. A suitable carrier may be selected from buffers, such as phosphate buffered saline (PBS) and tris buffered saline (TBS), and / or stabilizing agents such as sodium azide and thimerosal. Furthermore, peptides can be lyophilized (freeze-dried) and stored as a dry powder with or without the addition of stabilizers like mannitol or trehalose before lyophilization. In addition, peptides may be stored in a way to preserve their stability and activity. Peptides may be stored at -20°c or -80°C to maintain stability over long periods. Peptides may be stored in dark or opaque containers to protect them from light, which can cause degradation. Peptides may be stored in airtight containers to prevent oxidation and moisture absorption.

[0288] Antigens

[0289] The peptides and the compositions provided herein are useful as antigens for the detection, quantification and / or binding of for EBNA381-452 derived antibodies that cross-react with CNS- derived proteins. In general, any of the peptides or compositions provided herein can be used as antigens. In the sense of the present invention this is useful to detect the presence of EBNA381-452 derived antibodies that cross-react with CNS-derived proteins, which will bind the peptide or composition of the present invention thereby allowing a prognosis of an autoimmune disease such as MS. In Example 1 this has been demonstrated by using the peptides of SEQ ID NOs: 1-53 individually or in a pool, i.e. composition, as antigens for an ELISA assay. Accordingly, the invention provides an antigen comprising or consisting of the peptide of the present invention, or comprising or consisting of the composition of the present invention.

[0290] Methods

[0291] The peptide, composition and antigen of the present invention is particularly useful for prognosis of a subject’s risk of contracting MS. Accordingly, the peptide, composition and / or antigen of the present invention can be used in methods for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS. In general, the peptide, composition and / or antigen of the present invention are used to detect and / or quantify the presence of EBNA381-452 derived antibodies in a biological sample of a subject. The presence of those in turn significantly correlates with the subject’s risk of contracting an autoimmune disease, such as MS within the next 1 to 4 years (see Example 1). Accordingly, the methods of the present invention can prognose a subject’s risk of contracting an autoimmune disease, such as MS.

[0292] One advantage of the present invention is that the methods allow for an early prognosis of a subject being at risk of contracting an autoimmune disease such as MS. This contrasts with a diagnosis of the corresponding disease which is usually achieved at a much later stage when symptoms appear. Example 1 demonstrate that the prognosis indicates the subject’s risk of contracting the disease 1- 4 years prior the corresponding diagnosis. This allows acting even before the disease manifests. This has implicit consequences for the affected subject, because the identification of the risk relates to the pre-clinical phase of the disease such as MS, i.e. before symptoms appear and thus can aid the decision making process of the attending physician regarding pre-clinical monitoring, early diagnosis and / or early treatment to prevent clinical manifestation of the disease, e.g. MS, i.e. MS that is symptomatic for the affected subject. It is crucial to recognize MS patients before currently available diagnostic criteria, which currently require clinical symptoms, in order to be able to intervene in good time. Initial progress has been made through studies with clinically asymptomatic patients, which coincidentally reveal MS typical MRI lesions, thus termed radiologically isolated syndrome (RIS). Preventing the onset of the disease - after EBV infection but before MRI lesions develop - through vaccination or other immune system interventions would not cure MS but could potentially prevent the onset of the disease altogether.

[0293] Early detection of subjects being at risk of developing an autoimmune disease such as MS provides an opportunity to intervene in the underlying pathophysiological processes. This can potentially prevent or delay clinical development of MS and can even prevent disease progression and accumulating disability through early immunomodulatory treatments or vaccination strategies. Such proactive measures could effectively halt the progression of the disease, such as MS, or prevent its manifestation altogether. Thus, the methods for early prognosis provided herein are advantageous as they allow for early risk assessment as well as disease prevent! on / managem ent via tight pre-clinical monitoring, early diagnosis and / or early treatment to prevent manifestation or delay the progression of an autoimmune disease such as MS.

[0294] Accordingly, the invention provides a method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising detecting and / or quantifying binding of antibodies to the peptide of the invention, or the composition of the invention, or the antigen of the invention in a biological sample of said subject.

[0295] The invention also provides a method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising:

[0296] (a) contacting a biological sample of said subject with the peptide of the present invention, or the composition of the invention, or the antigen of the invention,

[0297] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0298] Any of the methods for early prognosis provided herein may also be in vitro or ex vivo. For example, the methods can be performed on an in vitro or ex vivo sample of a subject. In some embodiments, the methods for early prognosis provided herein are in vitro or ex vivo. In some embodiments, the methods for early prognosis provided herein do not comprise a step of obtaining a biological sample of a subject.

[0299] Accordingly, the present invention provides an in vitro or ex vivo method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising: (a) contacting a biological sample of said subject with the peptide of the invention, or the composition of the invention, or the antigen of the invention, (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0300] As used herein “detecting and / or quantifying binding of antibodies to the peptide of the invention, or the composition of the invention, or the antigen of the invention” may interchangeably be used with “detecting and / or quantifying the presence of antibodies binding to the peptide of the invention, or the composition of the invention, or the antigen of the invention.

[0301] As used herein “detecting and / or quantifying binding of antibodies to the peptide, the composition or the antigen of the present invention” preferably refers to detecting and / or quantifying the antibody level or antibody titer binding to the peptide, the composition or the antigen of the present invention in a subject or a sample of the subject. The antibody level or antibody titer may be defined by optical density (OD). As used herein “optical density (OD)” refers to the the difference in absorbance between two measurements, preferably at a wavelength of = 450 nm and = 620 nm. The terms OD and AOD may be used interchangeably. The OD may be measured at a wavelength of X = 450 nm and X = 620 nm, e.g. using a Synergy HTX multi-mode reader (BioTek, USA) or the like. Furthermore, the difference in absorbance between 450 nm and 620 nm can be calculated, followed by the subtraction of a blank sample. The skilled person is capable of choosing appropriate settings, e.g. wavelengths, to use the OD for measuring an antibody titer.

[0302] As used herein “contacting” in general means that two compounds are being brought into close proximity so that a reaction can happen. In the context of contacting a biological sample of a subject with the peptide of the present invention, or the composition of the invention, or the antigen of the invention, the term “contacting” means that the peptide of the present invention, or the composition of the invention, or the antigen of the invention is brought into close proximity with the biological sample allowing binding / association of EBNA381-452 derived antibodies with the peptide of the present invention, or the composition of the invention, or the antigen of the invention.

[0303] The methods for early prognosis of the present invention are particularly useful since prognosis can indicate a subject’s risk of contracting an autoimmune disease before diagnosis. This has been demonstrated in Example 1, where a prognosis was successful 1-4 years prior to MS diagnosis. The time frame between risk prognosis and MS diagnosis may vary. In particular, the time frame may depend on the EBV seroconversion, i.e. the change from a seronegative to a seropositive condition. This time frame may be about 14-15 years between seroconversion and MS diagnosis. The time frame may be about 8 years between seroconversion and MS diagnosis. The time frame may be about 1-2 years between seroconversion and MS diagnosis. Thus, the term “early prognosis” and / or “early diagnosis” in the context of the methods of the present invention may refer to the time frame between EBV seroconversion and MS diagnosis and / or the occurrence of MS symptoms. Accordingly, any of the methods provided herein may be performed as soon as a subject has undergone EBV seroconversion and / or before prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject.

[0304] Accordingly, in some embodiments, the methods for early prognosis provided herein may be performed between EBV seroconversion and prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject.

[0305] Accordingly, in some embodiments, the methods for early prognosis provided herein may be performed at least about 14-15 years prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In some embodiments, the methods for early prognosis provided herein may be performed at least about 8 years prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In some embodiments, the methods for early prognosis provided herein may be performed at least about 1-2 years prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In some embodiments, the methods for early prognosis provided herein may be performed at least about 14-15 years prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In a preferred embodiment, the methods for early prognosis provided herein may be performed after EBV seroconversion has been detected in a subject. In a preferred embodiment, the methods for early prognosis provided herein may be performed as soon as EBV seroconversion has been detected in a subject, and / or as soon as a subject is seropositive for EBV.

[0306] Accordingly, in one embodiment, the methods for early prognosis provided herein may be performed at least about 4 to 1 year, such as about 4, about 3, about 2, or at least about 1 year prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In another embodiment the methods for early prognosis provided herein may be performed about 1 month to about 50 years, such as about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about

[0307] 11 months, about 12 months, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, about 10 years, about 11 years, about 12 years, about 13 years, about 14 years, about 15 years, about 16 years, about 17 years, about 18 years, about 19 years, about 20 years, about 21 years, about 22 years, about 23 years, about 24 years, about 25 years, about 26 years, about 27 years, about 28 years, about 29 years, about 30 years, about 31 years, about 32 years, about 33 years, about 34 years, about 35 years, about 36 years, about 37 years, about 38 years, about 39 years, about 40 years, about 41 years, about 42 years, about 43 years, about 44 years, about 45 years, about 46 years, about 47 years. about 48 years, about 49 years, or about 50 years, prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In a preferred embodiment, the methods for early prognosis provided herein may be performed about 1 month to about 10 years, such as about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about

[0308] 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years, prior to a diagnosis or manifestation of an autoimmune disease, such as MS, in a subject. In one embodiment, the methods for early prognosis provided herein can prognose a subject’s risk of contracting an autoimmune disease, such as multiple sclerosis (MS) about 1 month to about 50 years, such as about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, about 10 years, about 11 years, about 12 years, about 13 years, about 14 years, about 15 years, about 16 years, about 17 years, about 18 years, about 19 years, about 20 years, about 21 years, about 22 years, about 23 years, about 24 years, about 25 years, about 26 years, about 27 years, about 28 years, about 29 years, about 30 years, about 31 years, about 32 years, about 33 years, about 34 years, about 35 years, about 36 years, about 37 years, about 38 years, about 39 years, about 40 years, about 41 years, about 42 years, about 43 years, about 44 years, about 45 years, about 46 years, about 47 years, about 48 years, about 49 years, or about 50 years, prior to a diagnosis or manifestation of an autoimmune disease, such as MS. In a preferred embodiment, the methods for early prognosis provided herein can prognose a subject’s risk of contracting an autoimmune disease, such as MS, about 1 month to about 10 years, such as about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about

[0309] 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years, prior to a diagnosis or manifestation of an autoimmune disease, such as MS. In a further preferred embodiment, the methods for early prognosis provided herein can prognose a subject’s risk of contracting an autoimmune disease, such as MS, about 4 to 1 year, such as about 4, about 3, about 2, or at least about 1 year prior to a diagnosis or manifestation of an autoimmune disease, such as MS. The methods for early prognosis provided herein may also be performed after diagnosis or manifestation of an autoimmune disease, such as MS, in a subject, e.g. to evaluate the contribution of EBV infection to the disease.

[0310] In Example 1, the inventors have demonstrated the power of the peptide, composition and / or antigen of the present invention for early prognosis of MS. This can be extrapolated to other autoimmune diseases that are affected or caused or partially caused by EBNA381-452 derived antibodies. Such diseases are for example autoimmune diseases selected from myasthenia gravis (MG), systemic lupus erythematosus (SLE), or autoimmune encephalitis (AE). Accordingly, in one embodiment, the autoimmune disease of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease provided herein is selected from MS, MG, SLE, or AE. In a preferred embodiment, the autoimmune disease of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease provided herein is MS.

[0311] In general, the risk prognosis in the methods for early prognosis provided herein is performed by detecting the presence of EBNA381-452 derived antibodies using the peptide or composition of the present invention as an antigen for detection. Example 1 shows that the presence of EBNA381-452 derived antibodies binding to the peptide or composition of the present invention significantly correlates with a risk of contracting MS. This is indicated in Figure 1, which shows that EBNA381- 452 derived antibodies are predominantly present in MS patients, while being absent in healthy controls. Accordingly, the presence of presence EBNA381-452 derived antibodies, which can be detected by the peptide or composition or antigen of the present invention and thereby indicates that a subject will contract MS. Accordingly, in one embodiment of the methods for early prognosis provided herein, the subject is at risk of contracting an autoimmune disease, such as MS, if binding of antibodies to the peptide, composition or antigen of the invention is detected and / or quantified in the biological sample of the subject. In another embodiment of the methods for early prognosis provided herein, the subject is at risk of contracting an autoimmune disease, such as MS, if the presence of antibodies binding to the peptide, composition or antigen of the invention is detected and / or quantified in the biological sample of the subject.

[0312] Accordingly, in one embodiment of the methods for early prognosis provided herein, the subject is not at risk or specific risk of contracting the autoimmune disease if no binding of antibodies to said peptide, composition or antigen is detected and / or quantified. In another embodiment of the methods for early prognosis provided herein, the subject is not at risk or specific risk of contracting the autoimmune disease if absence of antibodies binding to said peptide, composition or antigen is detected and / or quantified.

[0313] In general, quantification of the binding of antibodies to the peptides of the present invention is optional. This is because the detection of the presence of antibodies to the peptides of the invention is sufficient for prognosis. Already the presence of antibodies to the peptides of the present invention can be indicative for the subject to be at risk of contracting an autoimmune disease, such as MS. Quantification may further be used to fine tune the methods for prognosis provided herein. Accordingly, in any of the methods for early prognosis provided herein, quantification of binding of antibodies to the peptide, composition or antigen of the present invention is optional.

[0314] The methods for early prognosis provided herein may further comprise step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control. This has the advantage, that confidence of the prognosis can be enhanced. For example, a positive control can be used to verify the presence of antibodies binding to the peptide, composition or antigen of the invention in a subject, wherein a negative control can be used to verify the absence of antibodies binding to the peptide, composition or antigen of the invention in a subject. This can be realized by comparing values obtained from detecting and / or quantifying binding of antibodies in a biological sample of a subject with the corresponding values obtained from a positive or negative control. A positive and negative control may also be used to verify the functionality of the methods for early prognosis provided herein when performed on a sample of a subject and may thus be used as a quality control. Accordingly, in one embodiment, the methods for early prognosis provided herein may further comprise step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control.

[0315] The negative control may also be a value, for example the negative control can be a value indicating the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the present invention in a biological sample of one or more healthy subject. This negative control corresponds to a value of antibody binding in a sample of a subject which is not at risk of contracting an autoimmune disease, such as MS. As used herein “healthy subject” refers to an EBNA-1 -seropositive subject, without a diagnosis and history of autoimmune diseases, such as MS, and neuro-inflammatory disorders. As used herein “positive control” refers to a the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the present invention in a biological sample of one or more subjects that have contracted the autoimmune disease, preferably the one or more subjects are EBNA-1 -seropositive subjects with a diagnosis of an autoimmune disease, such as MS, or neuro-inflammatory disorders.

[0316] The positive control may also be a value, for example the positive control can be a value indicating the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the present invention in a biological sample of one or more subject that has contracted an autoimmune disease. This positive control corresponds to a value of antibody binding in a sample of a subject which is at risk of contracting an autoimmune disease, such as MS, or has already contracted the same. As used herein “subject that has contracted an autoimmune disease” refers to an EBNA-1- seropositive subject, with a diagnosis of an autoimmune disease, such as MS. The diagnosis of MS in said subject is preferably performed according to the McDonald criteria7As used herein, the term “diagnosis of MS” and the like refer to the identification of MS as the illness of a subject by examination of the symptoms, preferably according to the McDonald criteria7. The timepoint of diagnosis of MS refers to the timepoint at which the diagnosis of MS in a subject is completed, preferably according to the McDonald criteria7.

[0317] Accordingly, in one embodiment of the methods for early prognosis provided herein the method further comprises step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control, wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more healthy subject (negative control), and / or wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more subjects that have contracted the autoimmune disease (positive control). In some embodiments, the positive control is one or more biological samples of one or more healthy subjects. In general, the biological samples referred herein are biological samples of single subjects. It is preferred herein that biological samples of subjects are processed individually rather than pooled. This allows for prognosis of a subject-specific risk of contracting an autoimmune disease, such as MS. In some embodiments, the positive control is a value of detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention obtained from one or more biological samples of one or more healthy subject. In some embodiments, the negative control is a biological sample of one or more subject that has contracted an autoimmune disease, such as MS. In some embodiments, the negative control is a value of detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention obtained from a biological sample of one or more subject that has contracted an autoimmune disease.

[0318] The methods for early prognosis provided herein may comprise a step of comparing the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the present invention to a positive and / or negative control. This can indicate the subject’s risk of contracting an autoimmune disease, such as MS. For this, the methods for early prognosis may be performed on a biological sample of a subject and on a positive and / or negative control. The methods for early prognosis may also be performed only on a biological sample of a subject, wherein the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention is then compared to the corresponding value of a positive or negative control obtained earlier.

[0319] In general, it is indicative for a subject to be at risk of contracting an autoimmune disease, such as MS if the detected and / or quantified binding in a biological sample of the subject deviates from the negative control. For example, when binding of antibodies to the peptide, composition or antigen of the present invention is detected and / or quantified in a biological sample of a subject, wherein it is not detected and / or quantified in the negative control, said subject may be at risk of contracting an autoimmune disease, such as MS. Accordingly, the subject may not be at risk of contracting an autoimmune disease, such as MS, if binding of antibodies to the peptide, composition or antigen of the present invention is not detected and / or quantified in a biological sample of the subject consistent with the negative control, or is only detected and / or quantified in a biological sample of the subject to the same extend as in the negative control. For example, a subject, may be at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in the biological sample of the subject is increased compared to the negative control. If the detected and / or quantified binding in a biological sample of the subject deviates from the corresponding value in a negative control, i.e. is not to the same extend as in the negative control and / or is increased or decreased compared to the negative control, may be assessed by statistical tests known in the art. In this regard it is preferred, that the subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in a biological sample of the subject significantly deviates from a negative control, more preferably wherein the detected and / or quantified binding in a biological sample of the subject is significantly increased compared to the negative control. As used herein “significantly” refers to the statistical significance of a result that is unlikely to have occurred by chance alone, according to a predefined threshold of probability called the significance level, usually set at least to 0.05 or 5% or less. The skilled person is aware that statistical tests known in the art are necessary to allow a confident statement over significance of results / values.

[0320] Accordingly, in one embodiment of the methods for early prognosis provided herein, a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject significantly deviates from a negative control. In another embodiment of the methods for early prognosis provided herein, a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject is significantly increased compared to a negative control.

[0321] Accordingly, in one embodiment of the methods for early prognosis provided herein, a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject does not significantly deviate from a negative control. In another embodiment of the methods for early prognosis provided herein, a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject is significantly decreased compared to a negative control.

[0322] In a further embodiment, of the methods for early prognosis provided herein, a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject significantly deviates from the negative control, e.g. is increased compared to the negative control, and / or wherein a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in said biological sample of said subject does not significantly deviate from the positive control or is increased compared to the positive control.

[0323] In a further embodiment, of the methods for early prognosis provided herein, a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject does not significantly deviate from the negative control or is decreased compared to the negative control, and / or wherein a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in said biological sample of said subject does significantly deviate from the positive control, e.g. is decreased compared to the positive control.

[0324] In general, it is indicative for a subject to be at risk of contracting an autoimmune disease, such as MS if the detected and / or quantified binding in a biological sample of the subject does not deviate from the positive control. For example, when binding of antibodies to the peptide, composition or antigen of the present invention is detected and / or quantified in a biological sample of a subject, wherein it is also detected and / or quantified or wherein it is also detected and / or quantified to the same extend as in the positive control, the subject may be at risk of contracting an autoimmune disease, such as MS. Accordingly, the subject may not be at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the present invention in a biological sample of the subject does deviate from the positive control, e.g. is decreased compared to the positive control. For example, a subject, may not be at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in the biological sample of the subject is decreased compared to the positive control. A subject may be at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in the biological sample of the subject is increased compared to the positive control. If the detected and / or quantified binding in a biological sample of the subject corresponds to the value in a positive control, i.e. is to the same extend as in the positive control and / or is increased or decreased compared to the positive control, may be assessed by statistical tests known in the art. In this regard it is preferred, that the subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in a biological sample of the subject does not significantly deviate from a positive control, more preferably wherein the detected and / or quantified binding in a biological sample of the subject corresponds to the positive control and / or is increased compared to the positive control. In general, a detected and / or quantified binding that is decreased compared to the positive control may still be indicative for a subject’s risk of contracting an autoimmune disease such as MS, as long as the detected and / or quantified binding does not correspond to the negative control and / or is increased compared to the negative control. As used herein “deviates” means that two values are distinct from one another, e.g. the detected and / or quantified binding in a biological sample of a subject versus the corresponding values of a positive or negative control. It is preferred herein that the distinction of said values is significant, as explained above.

[0325] Accordingly, in one embodiment of the methods for early prognosis provided herein, a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject does not significantly deviate from a positive control. In another embodiment of the methods for early prognosis provided herein, a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject is significantly increased compared to a positive control.

[0326] Accordingly, in one embodiment of the methods for early prognosis provided herein, a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject does significantly deviate from a positive control. In another embodiment of the methods for early prognosis provided herein, a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of the subject is significantly decreased compared to a positive control.

[0327] In some embodiments, a deviation or significant deviation from the positive control is an increase or decrease in the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the positive control. In general, an increase of the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the positive control indicates, that the subject is at risk of contracting an autoimmune disease, such as MS. A decrease of the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the positive control may indicate, that the subject is not at risk of contracting an autoimmune disease, such as MS. However, as mentioned above, a subject may still be at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in a biological sample of the subject is significantly decreased compared to a positive control. This is because even a significant decrease in detected and / or quantified binding compared to a positive control may be an increase compared to a negative control. Thus it is preferred herein that a subject is not at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in a biological sample of the subject does not significantly deviate from a negative control or is even decreased compared to a negative control. Vice versa, it is preferred herein, that a subject is at risk of contracting an autoimmune disease, such as MS, if the detected and / or quantified binding in a biological sample of the subject does significantly deviate from a negative control and is increased compared to a negative control.

[0328] If no deviation or significant deviation of the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the positive control occurs, the subject may be at risk of contracting an autoimmune disease, such as MS.

[0329] In some embodiments, a deviation or significant deviation from the negative control is an increase or decrease in the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the negative control. In general, an increase of the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the negative control indicates, that the subject is at risk of contracting an autoimmune disease, such as MS. A decrease of the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the negative control indicates, that the subject is not at risk of contracting an autoimmune disease, such as MS. If no deviation or significant deviation of the detected and / or quantified binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject compared to the negative control occurs, the subject is not at risk of contracting an autoimmune disease, such as MS.

[0330] Accordingly, in one embodiment of the methods for early prognosis provided herein, a significant deviation from the positive control is an increase or decrease in the detected and / or quantified binding in said biological sample of said subject compared to the positive control. In one embodiment, the subject is at risk of contracting an autoimmune disease, such as MS, if the significant deviation from the positive control is an increase in the detected and / or quantified binding in said biological sample of said subject compared to the positive control. In another embodiment, the subject is not at risk of contracting an autoimmune disease, such as MS, if the significant deviation from the positive control is a decrease in the detected and / or quantified binding in said biological sample of said subject compared to the positive control.

[0331] In another embodiment of the methods for early prognosis provided herein, a significant deviation from the negative control is an increase or decrease in the detected and / or quantified binding in said biological sample of said subject compared to the negative control. In one embodiment, the subject is at risk of contracting an autoimmune disease, such as MS, if the significant deviation from the negative control is an increase in the detected and / or quantified binding in said biological sample of said subject compared to the negative control. In another embodiment, the subject is not at risk of contracting an autoimmune disease, such as MS, if the significant deviation from the negative control is a decrease in the detected and / or quantified binding in said biological sample of said subject compared to the negative control.

[0332] The methods for early prognosis provided herein are particularly advantageous, since they allow the medical practitioner, based on the prognosis, to make an informed, individualized decision on the further monitoring and / or treatment of the subject, e.g. a human patient. The clinical value of the present invention is the prognosis of the later development of an autoimmune disease, such as MS. This has implicit consequences for the affected subject, because the identification of the risk relates to the pre-clinical phase of the autoimmune disease, such as MS, and thus can determine the medical practitioner’s decisions regarding monitoring, e.g. clinical neurological monitoring and monitoring via MRI, or the earliest possible therapy to prevent the clinical manifestation of the autoimmune disease, such as MS, i.e. preventing symptomatic progression of the subject. This differs substantially current methods supporting diagnosis of an autoimmune disease, such as MS. At the point in time at which those methods come into play, the affected subjects a) are either already clinically neurologically symptomatic otherwise they would not seek medical clarification, or b) have a so-called Radiologically Isolated Syndrome (RIS). RIS corresponds to an incidental MRI finding usually during an MRI examination due to, for example, a concussion, headache or fear of a brain tumor with MRI changes that are typical of an autoimmune disease, such as MS. However, those affected do not yet have any specific neurological complaints.

[0333] In the case of a) prognosis and early therapeutic intervention is not possible since the subject is already symptomatic and thus will likely have a corresponding diagnosis.

[0334] In the case of b) the situation is of little help in the actual risk assessment of a future autoimmune disease, such as MS, manifestation, because the risk in RIS is predominantly due to spinal MRI changes, which in turn makes it unlikely that someone affected will actually have a spinal MRI without any neurological complaints i.e. at least already has subtle neurological complaints and therefore no longer has RIS.

[0335] The present invention overcomes this unsatisfying situation insofar, as the early prognosis provides an indication to the medical practitioner to initiate further monitoring or treatment steps for the affected subject, e.g. a human patient. In particular, the early prognosis also allows for an earlier diagnosis of an autoimmune disease, such as MS. An earlier diagnosis in turn facilitates the timely initiation of treatment, even before a clinically and MRI-based diagnosis of an autoimmune disease such as, MS or RIS, allows medical intervention, even before the onset of symptoms, as mentioned above. Early detection of subjects at risk of developing an autoimmune disease, such as MS, or RIS provides an opportunity to intervene in the underlying pathophysiological processes. This can potentially prevent the clinical manifestation of an autoimmune disease, such as MS, and avoid late diagnosis or further disease course through immunomodulatory treatments or vaccination strategies. Such proactive measures could effectively halt the progression of the disease or prevent its manifestation altogether. As such, the technical problem of the present invention may also be formulated as the provision of means and methods for the early prognosis of a future autoimmune disease, such as MS, by diagnosis of a subject.

[0336] The early prognosis achieved by the present invention, which has been shown to be successful in predicting a subject’s risk of contracting MS as early as 4 years before diagnosis (see Example 1), provides unmatched opportunities for disease management, and disease prevention. The focus after having achieved a prognosis, e.g. that a subject is at risk of contracting an autoimmune disease, such as MS, in the next years, is on monitoring the subject to allow early diagnosis of the disease and / or early therapy to avoid progression and manifestation of the disease.

[0337] Accordingly, in one embodiment of the methods for early prognosis provided herein, the methods further comprise advising pre-clinical monitoring to void symptomatic manifestation of an autoimmune disease, such as MS, and / or to allow early diagnosis of the autoimmune disease, e.g. MS, if the subject is at risk of contracting the autoimmune disease.

[0338] A diagnosis of an autoimmune disease of the present invention is explained in the following exemplary for MS. Diagnosis of MS typically is initiated with clinical neurological examination following symptoms, indicated by e.g. visual disturbances, sensory sensations, difficulty walking, and motor impairments, a neurological assessment is conducted to identify existing impairments. This is followed by imaging of the brain via MRI and, if needed, the spinal cord. Based on clinical examinations and MRI findings, a tentative diagnosis of MS is made. A lumbar puncture may be advisable to exclude alternative diagnoses and detect chronic inflammatory activity in the cerebrospinal fluid (CSF). While not mandatory, many specialized organizations recommend it. The diagnosis of MS follows the principle of spatial and temporal dissemination (i.e. restrictions at two points in time in two functional systems - e.g. brainstem and visual system). Currently, according to the latest diagnostic criteria (although new ones are due to be introduced later this year), at least one clinical symptom is required. In addition, MRI assessment can reveal lesions in different topographical CNS areas indicating dissemination in space, with active lesions emphasized by contrast-agent enhancement and older lesions helping to confirm dissemination in time. In cases without contrast-agent enhancement, dissemination in time can be confirmed by the detection of CSF oligoclonal bands or intrathecal immunoglobulin synthesis, indicating chronic inflammation in the CSF. It is noted that these are the currently valid diagnosis criteria of MS which are regularly revised. For example, revised diagnosis criteria for MS will most likely be presented in September 2024 and might differ from the above. It is preferred herein that MS is diagnosed according to the most recent revision of the diagnosis criteria of MS.

[0339] Clinically isolated syndrome (CIS) is a condition that does not directly lead to an MS diagnosis as it indicates a lack of dissemination in time or space. This may manifest as a single clinical lesion within a functional system and thus not fulfil the criteria for dissemination in time and space or as multiple MRI lesions detected at a single time point and thus not fulfil the criteria for dissemination in time. In general, CIS has become very rare. CIS is described in detail in Lublin et al.22which is incorporated herein in its entirety.

[0340] Radiologically isolated syndrome (RIS) is a further condition that does not directly lead to an MS diagnosis as it describes a scenario in which the typical clinical symptoms suggestive of MS are absent, but MRI shows characteristic lesions. However, depending on risk factors such as the presence of CSF oligoclonal bands or spinal MRI lesions, or a prognosis in the sense of the present invention, there is an increased likelihood that MS will develop.

[0341] In all of these scenarios, the focus is on relapsing-remitting disease characterized by worsening symptoms at clearly identifiable times. However, if there is a gradual worsening over time without clearly identifiable time points and typical symptoms are present in conjunction with MRI lesions, a diagnosis of progressive (primary) MS may be warranted.

[0342] It is noteworthy that in the majority of patients, relapsing-remitting MS transitions into a progressive disease course, known as secondary progressive MS, in which the progressive phase follows a relapsing phase. Recent studies suggest that treatment may influence both the timing and likelihood of transition to the progressive phase.

[0343] In the majority of patients, multiple lesions are observed at the time of diagnosis, suggesting that radiological activity has taken place before patients have become symptomatic. There is a time lag between the onset of the first symptoms and the confirmed diagnosis, although advances such as the availability of MRI and updated diagnostic criteria allow earlier detection. However, there is a “silent” period prior to the appearance of the first symptoms, i.e. a period of time where lesions appear but no symptoms and thus no diagnosis as the subject will usually not see a medical practitioner in this time. This time period can be crucial to start early treatment. The methods for early prognosis of the present invention allow to prognose a subject’s risk of contracting an autoimmune disease, such as MS, and the likelihood of a corresponding diagnosis in the feature. Thus, the invention allows to use the above-mentioned time period to allow earlier diagnosis and / or initiate early treatment, e.g. to avoid axonal damage. This offers the possibility of delaying the transition from an asymptomatic subject to a symptomatic subject or may even allow halting the progression of MS diagnosis with early treatment. The early diagnosis and / or early treatment of the subject can be achieved by tight pre-clinical monitoring of the subject after a corresponding risk prognosis.

[0344] Monitoring of a subject that has been prognosed to be at risk of contracting an autoimmune disease, such as MS, can include pre-clinical monitoring in the form of frequent medical examination, neurological and radiological monitoring. Neurological and radiological monitoring can include MRI, lumbar puncture, optical coherence tomography (OCT) and / or electrophysiological examination. MRI for example can be used to detect lesions, e.g. in the brain and / or spinal cord of a subject. Lumbar puncture can be used to detect chronic inflammatory activity in the CSF of a subject. The aim of such pre-clinical monitoring is to assess the transition of an asymptomatic subject, to a symptomatic subject which should then immediately be diagnosed and commence treatment.

[0345] Accordingly, in one embodiment of the methods for early prognosis provided herein, the methods further comprise advising pre-clinical monitoring, wherein the pre-clinical monitoring comprises frequent medical examination, such as neurological and radiological monitoring of the subject, e.g. via MRI of the subject’s brain and / or spinal cord, lumbar puncture, OCT and / or el ectrophy si ol ogi cal examinati on .

[0346] Early treatment of a subject that has been prognosed to be at risk of contracting an autoimmune disease, such as MS, can be particularly beneficial for avoiding manifestation of the disease and / or delaying the progression of the disease. Treatment may be advised to a subject that has been prognosed to be at risk of contracting an autoimmune disease, such as MS in accordance with the present invention. Accordingly, in one embodiment of the methods for early prognosis provided herein, the methods further comprise advising treatment to the subject. In another embodiment of the methods for early prognosis provided herein, the methods further comprise treating the subject. In general, the treatment can comprise any treatment to intervene with the underlying pathophysiological processes. For example, the treatment can comprise treatment with immunosuppressants or vaccination.

[0347] Timely treatment of MS is critical to limit axonal damage and prevent disability. Several mildly to moderately effective treatments are available, including interferon beta drugs (Rebif, Betaseron, Extavia, Plegredy, Avonex, and biosimilars), glatiramer acetate (Copaxone and follow-on drugs), teriflunomide (Aubagio and generics) and dimethyl fumarate / diroximel fumarate (Tecfidera / Vumerity and generics). These medications may be started before or after a diagnosis of MS, although interferon preparations are also suitable for CIS patients with indication of MRI dissemination in time and space. While these drugs have a mild to moderate effect on disease activity, recent advances have expanded treatment options.

[0348] Currently, moderate-highly effective treatments of MS such as SIP modulators (Fingolimod, Zeposia, Ponvory and biosimilars) or cladribine (Mavenclad) can be used depending on individual benefit-risk evaluation. Although traditionally seen as escalating treatments for cases where mildly to moderately effective drugs are ineffective, evolving treatment strategies recognize the impact of early and severe disease activity on long-term disability. Most notably, the SIP modulator Mayzent has been approved for CIS, relapsing and active SPMS, representing a paradigm shift in treatment.

[0349] Highly effective treatments of MS are natalizumab (Tysabri and biosimilars), CD20 antibodies (ocrelizumab - Ocrevus, ofatumumab - Kesimpta, ublituximab - Briumvi, and off-label rituximab). These can be used as escalation treatment or even as initial therapy in highly active disease characterized by either relapses despite previous medication, or frequent relapses and high inflammatory MRI activity, i.e. increasing or enlarging number of T2 lesions.

[0350] Alemtuzumab (Lemtrada) is restricted due to significant side effects to MS cases with highly active disease that have not responded to previous treatments.

[0351] In one aspect of the invention, early treatment of an autoimmune disease, such as MS, comprises treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon P-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0352] In some embodiments, the early treatment of an autoimmune disease, such as MS, is a diseasemodifying treatment and comprises a treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon P-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Ublituximab (Briumvi) and the like or any combinations thereof.

[0353] In some embodiments, the early treatment of an autoimmune disease, such as MS, is a symptomatic treatment and comprises a treatment with a drug selected from the list consisting of Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0354] The early treatment of a subject that is at risk of contracting an autoimmune disease, such as MS, according to the methods for early prognosis of the invention, may also comprise weakening of the subject’s immune system by therapy in order to prevent the misdirected overreaction of the subject’s immune system triggered by EBV and in particular EBNA381-452. The closest to this case is currently RIS, in which there are no clinical symptoms yet, but lesions are already recognizable in the MRI, so MS symptoms could probably occur in the future. The prognosis thus represents an indication for early treatment of an autoimmune disease, such as MS. Lebrun-Frenay et al. 202320 demonstrates that early treatment can delay the appearance of the first clinical symptoms.

[0355] Accordingly, the invention provides in one aspect method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising detecting and / or quantifying binding of antibodies to the peptide, or the composition, or the antigen of the invention in a biological sample of said subject, wherein the method further comprises advising early treatment if the subject is at risk of contracting the autoimmune disease.

[0356] The invention also provides a method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising:

[0357] (a) contacting a biological sample of said subject with the peptide, or the composition, or the antigen of the invention,

[0358] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen, wherein the method further comprises advising early treatment if the subject is at risk of contracting the autoimmune disease.

[0359] The invention also provides an in vitro or ex vivo method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising:

[0360] (a) contacting a biological sample of said subject with the peptide, or the composition, or the antigen of the invention,

[0361] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen, wherein the method further comprises advising early treatment if the subject is at risk of contracting the autoimmune disease.

[0362] In one embodiment, the step of advising early treatment comprises advising treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0- 1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0363] In some embodiments, the step of advising early treatment comprises advising disease-modifying treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0- 1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Ublituximab (Briumvi) and the like or any combinations thereof.

[0364] In some embodiments, the step of advising early treatment comprises advising symptomatic treatment with a drug selected from the list consisting of Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0365] In another embodiment, the treatment comprises administering Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0-1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof to the subject.

[0366] In some embodiments, the treatment is a disease-modifying treatment and comprises administering Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0-1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0367] In some embodiments, the treatment is a symptomatic treatment and comprises administering Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0368] In one aspect of the invention, the methods for early prognosis provided herein do not comprise a treatment step.

[0369] In another aspect of the invention, the methods for early prognosis provided herein may comprise a further treatment step as defined above in the context of advising early treatment.

[0370] Accordingly, the invention provides in one aspect method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising detecting and / or quantifying binding of antibodies to the peptide, or the composition, or the antigen of the invention in a biological sample of said subject, wherein the method further comprises a treatment of the autoimmune disease if the subject is at risk of contracting the autoimmune disease.

[0371] The invention also provides a method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably MS, the method comprising:

[0372] (a) contacting a biological sample of said subject with the peptide, or the composition, or the antigen of the invention,

[0373] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen, wherein the method further comprises a treatment of the autoimmune disease if the subject is at risk of contracting the autoimmune disease.

[0374] In one embodiment, the step of treatment of the autoimmune disease comprises treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon -lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0375] In some embodiments, the step of treatment of the autoimmune disease comprises a diseasemodifying treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0- 1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0376] In some embodiments, the step of treatment of the autoimmune disease comprises symptomatic treatment with a drug selected from the list consisting of Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0377] In another embodiment, the treatment of the autoimmune disease comprises administering Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0-1 a (Avonex, Rebif), Interferon 0- lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), or Vardenafil (Levitra, Staxyn, and others) and the like, or any combinations thereof to the subject. In some embodiments, the treatment of the autoimmune disease is a disease-modifying treatment and comprises administering Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0-1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0378] In some embodiments, the treatment of the autoimmune disease is a symptomatic treatment and comprises administering Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0379] As used herein “disease-modifying treatment” relates to an early treatment, e.g. after prognosis and before a subject has become symptomatic, e.g. has been diagnosed with an autoimmune disease such as MS.

[0380] As used herein “symptomatic treatment” relates to a treatment after a subject has become symptomatic, e.g. has been diagnosed with an autoimmune disease. Both disease-modifying and symptomatic treatments can be applied after a subject has been prognosed to be at risk of contracting an autoimmune disease such as MS in the sense of the present invention.

[0381] The present invention is particularly advantageous since the detection of a subject’s risk of contracting an autoimmune disease, such as MS, or the diagnosis of a risk of contracting an autoimmune disease, such as MS, allows to alter the standard treatment to benefit the subject’s prognosis. For example, the above-described treatment of MS usually starts when MS is diagnosed in a subject. However, an MS diagnosis is usually late due to the differential character. This means that subjects may already have suffered lesions, e.g. in the brain and / or spinal cord, resulting in permanent damage and disability. The methods and uses of the present invention allow to alter this standard MS treatment by providing a treatment for early immune suppression, e.g. initiating immunosuppressive therapy before lesions in the brain and / or spinal cord are detectable, thereby preventing the formation of sclerosis lesions ultimately averting the development of MS. The early immune suppression may also be combined with strict monitoring of patients being at risk of contracting MS as described above. Rigorous follow-up monitoring of the subject’s enables the medical practitioner to detect sclerosis lesions early before symptoms occur, e.g. via MR- (Radiologically Isolated Syndrome - RIS). This early detection can then be the starting point for early immunosuppressive therapy thereby inhibiting the progression of sclerosis lesions and manifestation of MS. Both strategies allow for a treatment of MS before diagnosis, thereby providing novel treatment strategies benefitting patients at risk of contracting MS.

[0382] Furthermore, if a subject is identified being at risk of contracting an autoimmune disease, such as MS, a novel treatment strategy may be vaccination against EBV. There are no EBV vaccines approved so far, however, considering the prognosis of patients being at risk of contracting e.g. MS, a further novel treatment strategy is to conduct vaccination trials on said patients to prevent or reduce EBV derived autoantibodies that contribute to the manifestation of MS in the patients. An exemplary EBV vaccine that can be used in this context is disclosed in WO 2024 / 084091 which is incorporated herein in its entirety.

[0383] Furthermore, the present invention allows for better personalized treatment via a personalized MS prognosis of a subject in years. As shown in Example 4 the present invention can assess a subject’s risk of contracting MS starting from EBV- seroconversion in years, i.e. the detected and / or quantified antibody titer in a subject or a sample from the subject measured at several timepoints after EBV seroconversion can indicate the timeframe in which the subject most likely will contract MS. This allows also for a delay of early treatment. For example, the treatment may be started once the subject is most likely to contract MS, in the case of a subject that has been detected and / or quantified with a high antibody titer four out of four times this would mean to start a treatment about 3 years after EBV seroconversion. This is demonstrated in Fig.161 of Example 4.

[0384] Accordingly, in one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the method further comprises advising a delay of treatment of the autoimmune disease, if the subject is at risk of contracting the autoimmune disease in about 1 to about 20 years, such as 1, 2, 3, 4 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years after EBV seroconversion, preferably in about 3 to about 6, such as 3, 4, 5, or 6 years after EBV seroconversion, preferably in about 3 to about 13, such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 years after EBV seroconversion, preferably in about 8 to about 13, such as 8, 9, 10, 11, 12, or 13 years after EBV seroconversion.

[0385] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the method further comprises delaying treatment of the autoimmune disease, if the subject is at risk of contracting the autoimmune disease in about 1 to about 20 years, such as 1, 2, 3, 4 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years after EBV seroconversion, preferably in about 3 to about 6, such as 3, 4, 5, or 6 years after EBV seroconversion, preferably in about 3 to about 13, such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 years after EBV seroconversion, preferably in about 8 to about 13, such as 8, 9, 10, 11, 12, or 13 years after EBV seroconversion.

[0386] The invention is not particularly limited to any methods for detecting and / or quantifying binding of antibodies to the peptide, composition or antigen of the present invention. Such methods are known in the art. In general, any methods that can detect the presence and / or quantify the presence of an antibody binding to the peptide, composition or antigen of the present invention may be used. Exemplary assays to detect and / or quantify binding of antibodies to the peptide, composition or antigen of the present invention are Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC), Flow Cytometry and / or Fluorescence-Activated Cell Sorting (FACS) and the like. In a sandwich ELISA (enzyme-linked immunosorbent assay) or similar formats of immunoassay, a “sandwich” is formed by the antibody of interest being trapped between two peptide molecules - one immobilized on a solid phase and another attached to a detection system.

[0387] In a preferred embodiment, binding of antibodies to the peptide, composition or antigen of the present invention is detected and / or quantified via ELISA. Accordingly, in one embodiment of the methods for early prognosis provided herein, binding of antibodies is detected and / or quantified using Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC), Flow Cytometry and / or Fluorescence-Activated Cell Sorting (FACS) and the like.

[0388] The methods for early prognosis provided herein are performed on a biological sample of a subject. In some embodiments of the methods for early prognosis provided herein, the methods do not comprise a step of obtaining a biological sample of a subject. In some embodiments of the methods for early prognosis provided herein, the biological sample has already been obtained before performing the method, i.e. is not part of the method. In a preferred embodiment of the methods for early prognosis provided herein, the biological sample is an in vitro or ex vivo sample.

[0389] A biological sample refers to any substance taken from a living organism or from the environment where living organisms are found. A biological sample is preferably a human biological sample. A biological sample may comprise blood or blood components, such as blood serum and / or blood plasma, tissues from biopsies or surgeries, cells collected from swabs e.g. cheek cells and / or cervical cells, body fluids such as saliva, urine, CSF, semen, amniotic fluid, fecal matter and the like. A biological sample is preferably a blood sample. A biological sample is preferably a whole blood sample, a blood serum sample, or a blood plasma sample. A biological sample is most preferably a human biological sample, a human biological blood sample, a human blood sample, a human whole blood sample, a human blood serum sample, and / or a human blood plasma sample. Accordingly, in one embodiment of the methods for early prognosis provided herein, the biological sample is a blood sample or a CSF sample. In a further embodiment of the methods for early prognosis provided herein, the blood sample is a whole blood sample, a blood serum sample, or a blood plasma sample. In a further embodiment of the methods for early prognosis provided herein, the biological sample or blood sample or CSF sample is a human biological sample or a human blood sample or a human CSF sample.

[0390] In another aspect of the methods for early prognosis provided herein, the methods can further comprise a step of obtaining a biological sample of a subject.

[0391] As used herein, the term “subject” means any individual. A subject can be for example an animal that is susceptible to an autoimmune disease such as MS. In the sense of the present invention a subject is preferably a mammal. A subject is most preferably a human. A subject is preferably a human that has contracted an EBV infection and / or is EBV seropositive. A subject may be a human patient. A subject or patient may be characterized by the presence of EBNA381-452 peptide-specific IgG antibody titers in a biological sample of the subject or patient. In particular, Example 4 demonstrates that high-level EBNA38i-452-derived antibody titers are directly associated with the development of MS in a subject or patient. A specific level of EBNA38i-452-derived antibody titers can therefore be used to predict a subject’s risk of contracting MS in the future.

[0392] In one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the step of detecting and / or quantifying binding of antibodies to said peptide, composition or antigen comprises assessing the EBNA-138i-452-specific IgG antibody titer in the biological sample of the subject.

[0393] Accordingly, in one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, if the detected and / or quantified binding in the biological sample of the subject corresponds to an EBNA- 138i-452-specific IgG antibody titer of OD>1.7, such as an OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4, optionally wherein the OD is measured at a wavelength of = 450 nm and = 620 nm. In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is not at risk or at low risk of contracting the autoimmune disease, such as MS, if the detected and / or quantified binding in the biological sample of the subject corresponds to an EBNA- 138i-452-specific IgG antibody titer of OD<1.69, such as an OD<1.6, OD<1.5, OD<1.4, OD<1.3, OD<1.2, OD<1.1, OD<1, OD<0.9, OD<0.8, OD<0.7, OD<0.6, OD<0.5, OD<0.4, OD<0.3, OD<0.2, OD<0.1, or an ODA), optionally wherein the OD is measured at a wavelength of X = 450 nm and X = 620 nm.

[0394] The OD may be measured at a wavelength of X = 450 nm and X = 620 nm, e.g. using a Synergy HTX multi-mode reader (BioTek, USA) or the like. Furthermore, the difference in absorbance between 450 nm and 620 nm can be calculated, followed by the subtraction of the OD value of a blank sample.

[0395] The level of EBNA381 -452-derived antibody titers may be assessed in the subject or patient or a sample of the subject or patient at any time, preferably after EBV seroconversion. Example 4 demonstrates that the level of EBNA38i-452-derived antibody titers at any time after EBV seroconversion can indicate a subject’s risk of contracting MS. In one embodiment the level of EBNA38i-452-derived antibody titers in a subject or a sample of the subject may be measured after EBV seroconversion. In another embodiment, the level of EBNA38i-452-derived antibody titers may be measured after EBV seroconversion and up to 4 years, such as 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month(s), before disease onset or after EBV seroconversion. As used herein “disease onset” refers to the timepoint at which the autoimmune disease, such as MS, manifests, e.g. the timepoint at which first clinical symptoms appear, and / or is diagnosed, in the case of MS preferably by examination of the symptoms according to the McDonald criteria7. The timepoint at which first clinical symptoms of MS appear may be before or after a MS diagnosis, e.g. according to the 2024 revised version of the McDonald Diagnostic Criteria.

[0396] Accordingly, in one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the method is performed between about 10 years and about 1 month, such as about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 1 year, about 2 years, about 3 years, about 4 years, about 5 years, about 6 years, about 7 years, about 8 years, about 9 years, or about 10 years after Epstein-Barr virus (EBV) seroconversion in the subject.

[0397] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the method is performed between about 1 month and about 48 months, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, or 48 months after Epstein-Barr virus (EBV) seroconversion in the subject.

[0398] In addition to the level of EBNA38i-452-derived antibody titers in a subject or a sample of the subject measured at a single timepoint, Example 4 demonstrates that high levels of EBNA38i-452-derived antibody titers in a subject or a sample of the subject measured over a prolonged time period are associated with the development of MS. Accordingly, a high level of EBNA38i-452-derived antibody titers in a subject or a sample of the subject measured at several timepoints can indicate a subject’s risk of contracting an immune disease, such as MS. Thus, the level of EBNA381-452- derived antibody titers may be assessed in the subject or patient or a sample of the subject or patient at several timepoints over a period of time, preferably after EBV seroconversion. In one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, if the subject or a sample of the subject exhibits an EBNA-138i-452-specific IgG antibody titer of OD>1.7, such as an OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4, over a prolonged time period or in several measurements performed over a period of time after seroconversion, preferably, wherein the prolonged time period is up to 4 years, such as 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31,

[0399] 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month(s), before disease onset or after EBV seroconversion, or wherein the measurements are performed up to 4 years, such as 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32,

[0400] 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month(s), before disease onset or after EBV seroconversion.

[0401] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is not at risk or at low risk of contracting the autoimmune disease, such as MS, if the subject or a sample of the subject exhibits an EBNA-138i-452-specific IgG antibody titer of OD<1.69, such as an 0D<1.6, 0D<1.5, 0D<1.4, 0D<1.3, 0D<1.2, OD<1.1, OD<1, OD<0.9, OD<0.8, OD<0.7, OD<0.6, OD<0.5, OD<0.4, OD<0.3, OD<0.2, OD<0.1, or an OD=0, over a prolonged time period or in several measurements performed over a period of time after seroconversion, preferably, wherein the prolonged time period is up to 4 years, such as 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20,

[0402] 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month(s), before disease onset or after EBV seroconversion, or wherein the measurements are performed up to 4 years, such as 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22,

[0403] 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month(s), before disease onset or after EBV seroconversion.

[0404] As shown in Example 4, if a high level of EBNA38i-452-derived antibody titers in a subject or a sample of the subject is detected at multiple timepoints after EBV seroconversion and before disease onset, the subject is at a higher risk of contracting MS.

[0405] Accordingly, in one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, if the subject or a sample of the subject exhibits an EBNA-138i-452-specific IgG antibody titer of OD>1.7, such as an OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4 at 1-10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 measurements performed at different timepoints after EBV seroconversion. In a preferred embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, if the subject or a sample of the subject exhibits an EBNA-138i-452-specific IgG antibody titer of OD>1.7, such as an OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4 at 1-4, such as 1, 2, 3, or 4 measurements performed at different timepoints after EBV seroconversion. In a more preferred embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, if the subject or a sample of the subject exhibits an EBNA-I381- 452-specific IgG antibody titer of OD>1.7, such as an OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4 at2-4, such as 2, 3, or 4 measurements performed at different timepoints after EBV seroconversion.

[0406] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is not at risk or at low risk of contracting the autoimmune disease, such as MS, if the subject or a sample of the subject exhibits an EBNA-138i-452-specific IgG antibody titer of OD>1.7, such as an OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4 at none to 2, such as 0, 1, or 2 measurements performed at different timepoints after EBV seroconversion. In general, the measurements to define the OD of the EBNA-138i-452-specific IgG antibody titer in a subject or a sample of the subject may be performed at any time after EBV seroconversion, e.g. 1-48 months, such as 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 month(s) after EBV seroconversion in the subject or a sample of the subject. It is preferred, that the first measurement to define the OD of the EBNA-138i-452-specific IgG antibody titer in a subject or a sample of the subject may be performed about 1-12 months, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months after EBV seroconversion in the subject or a sample of the subject. The measurement may then be repeated regularly, preferably in periods of about 1-12 months, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, such that each measurement of the OD of the EBNA-I381-452- specific IgG antibody titer in a subject or a sample of the subject is performed within 1-12 months, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 of the last measurement. This is to ensure to detect episodes of high-level EBNA-138i-452-specific antibody levels in the subject or sample of the subject thereby increasing the confidentiality in the subject’s risk of contracting MS and the subjects expected disease onset.

[0407] In one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about every 1-12 months, such as every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 month(s).

[0408] In one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 times, preferably about 1, 2, 3 or about 4 times, after EBV seroconversion in the subject. When the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention are regularly performed on a sample of a subject, each time they are performed on a fresh biological sample of the subject to reflect the up to date condition of the subject.

[0409] As used herein “low risk” refers to a situation wherein an EBNA-138i-452-specific IgG antibody titer is detected and / or quantified in a biological sample of a subject, however, wherein said EBNA-138i-452-specific IgG antibody titer is detected and / or quantified in a biological sample of a subject at an OD of <1.69 i.e. that the presence of the EBNA-138i-452-specific IgG antibody titer does not indicate a significant association with contracting an autoimmune disease, such as MS in the future.

[0410] As used herein “not at risk” or “no risk” or “negative risk” preferably means that not EBNA-hsi- 452-specific IgG antibody titer can be detected and / or quantified in a biological sample of a subject, resulting in no significant association with the development of an autoimmune disease, such as MS.

[0411] As used herein “measurement” or “to measure” refers to detecting and / or quantifying binding of antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject, thereby assessing the EBNA-138i-452-specific IgG antibody titers in said biological sample in the context of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention. Furthermore, Example 4 demonstrates that the subjects having at least one episode of high-level EBNA-138i-452-specific antibody levels after EBV seroconversion are more likely to have an earlier MS disease onset. For example, the presence of four episodes of high-level EBNA-138i-452-specific antibody levels was associated with an early MS disease onset of only about 3 to 6 years after EBV seroconversion. In contrast, the absence or reduced number of such episodes of high-level EBNA- 138i-452-specific antibody levels was associated with a delayed disease onset. For Example, the presence of three and two episodes of high-level EBNA-138i-452-specific antibody levels is still associated with an early MS disease onset of about 3 to 13 years. Here, the time range of disease onset varies between subjects. Finally, the absence of episodes of high-level EBNA-I381-452- specific antibody levels is associated with a very delayed disease onset of about 8 to 13 years. It is noted that also subjects with no episodes of high-level EBNA-138i-452-specific antibody levels may contract MS after e.g. about 13 years as shown in Example 4, Figure 16 1. This is due to the retrospective experimental setup which only included MS patients. However, this does not mean that a subject having no episodes of high-level EBNA-138i-452-specific antibody levels or in general has a low level EBNA-138i-452-specific antibody titer of OD<1.69 can stay healthy, i.e. is not at risk or at low risk of contracting MS in the future. This demonstrates that the number of episodes of high-level EBNA-138i-452-specific antibody levels indicate the risk of an earlier disease outbreak of an autoimmune disease, such as MS, in a subject.

[0412] Accordingly, in one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, in about 1 to 10 years, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 years, preferably in about 3 to 6 years, such as 3, 4, 5, or 6 years after EBV seroconversion if the subject or a sample of the subject exhibits a high- level EBNA-138i-452-specific antibody titer at least about 4 individual measurements / episodes after EBV seroconversion.

[0413] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, in about 1 to 15 years, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 years, preferably in about 3 to 13 years, such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 years after EBV seroconversion if the subject or a sample of the subject exhibits a high-level EBNA-138i-452-specific antibody titer at at least about 2 or at least about 3, preferably 2 or 3 individual measurements / episodes after EBV seroconversion.

[0414] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease, such as MS, in about 5 to 20 years, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years, preferably in about 8 to 13 years, such as 8, 9, 10, 11, 12, or 13 years after EBV seroconversion if the subject or a sample of the subject does not exhibit a high-level EBNA-138i-452-specific antibody titer at any individual measurements / episodes after EBV seroconversion.

[0415] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is not at risk or at low risk of contracting the autoimmune disease, such as MS, in about 5 to 20 years, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 years, preferably in about 8 to 13 years, such as 8, 9, 10, 11, 12, or 13 years after EBV seroconversion if the subject or a sample of the subject does not exhibit a high-level EBNA-138i-452-specific antibody titer at any individual measurements / episodes after EBV seroconversion.

[0416] As used herein “high-level EBNA-138i-452-specific antibody” refers to an antibody level or antibody titer in a subject or a sample of the subject of OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or an OD>4, wherein the OD is measured at a wavelength of = 450 nm and = 620 nm. As used herein “low-level EBNA-138i-452-specific antibody” refers to an antibody level or antibody titer in a subject or a sample of the subject of OD<1.69, such as an OD<1.6, OD<1.5, OD<1.4, OD<1.3, OD<1.2, OD<1.1, OD<1, OD<0.9, OD<0.8, OD<0.7, OD<0.6, OD<0.5, OD<0.4, OD<0.3, OD<0.2, OD<0.1, or an OD=0, wherein the OD is measured at a wavelength of X = 450 nm and X = 620 nm.

[0417] In one embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention the subject is at risk of contracting the autoimmune disease in about 1 to 10 years, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 years, preferably in about 3 to about 6 years, such as 3, 4, 5, or 6 years, after EBV seroconversion if

[0418] (i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, times, preferably about 1, 2, or 3 times, most preferably about 4 times after EBV seroconversion in the subject, and

[0419] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, such as OD>1.7, OD>1.8, OD>1.9, 0D>2, 0D>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, 0D>3, 0D>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or OD>4, optionally, measured at a wavelength of = 450 nm and = 620 nm.

[0420] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is at risk of contracting the autoimmune disease in about 1 to about 15 years, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 years, preferably in about 3 to about 13 years, such as 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, or 13 years after EBV seroconversion if

[0421] (i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, times, preferably about 2 or 3 times after EBV seroconversion in the subject, and

[0422] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, such as OD>1.7, OD>1.8, OD>1.9, OD>2, OD>2.1, OD>2.2, OD>2.3, OD>2.4, OD>2.5, OD>2.6, OD>2.7, OD>2.8, OD>2.9, OD>3, OD>3.1, OD>3.2, OD>3.3, OD>3.4, OD>3.5, OD>3.6, OD>3.7, OD>3.8, OD>3.9, or OD>4, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0423] In another embodiment of the methods for early prognosis of a subject being at risk of contracting an autoimmune disease, such as MS, or the peptide, composition, or antigen for use of the invention, the subject is not at risk or at low risk of contracting the autoimmune disease in about 5 to about 20 years, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or20 years, preferably in about 8 to about 13 years, such as 8, 9, 10, 11, 12, or 13 years after EBV seroconversion if

[0424] (i) no binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified after EBV seroconversion in the subject, and / or

[0425] (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, such as an OD<1.6, OD<1.5, OD<1.4, OD<1.3, OD<1.2, OD<1.1, OD<1, OD<0.9, OD<0.8, OD<0.7, OD<0.6, OD<0.5, OD<0.4, OD<0.3, OD<0.2, OD<0.1, or OD=0, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

[0426] As used herein “human” includes humans of all ages and sex, for example female or male children, adults, or elderly people / person. Subjects in the sense of the present invention are for example subjects that may be at risk of contracting an autoimmune disease, such as MS, and / or have been exposed to EBV, have been infected with EBV or subjects that are at risk of EBV infection. A subject in the sense of the present invention may be at risk of contracting an autoimmune disease, such as MS, based on disease history in the family or other indications. In some embodiments, a subject may be predisposed for an autoimmune disease, such as MS. The methods for early prognosis of the invention may be performed on a biological sample of any subject, where it is desired to make a prognosis of the subject’s risk of contracting an immune disease, such as MS. A subject in the sense of the invention may also be a subject that has already contracted an immune disease, such as MS. This can be beneficial to identity the underlying pathological process of the disease, e.g. the presence of EBNA381-452 derived antibodies that cross-react with CNS-derived proteins. In some embodiments, the subject is EBNA-l-seropositive.

[0427] Diagnosis

[0428] As explained above, prognosis of the risk of a subject of contracting an autoimmune disease, such as MS, allows for early diagnosis of said disease. The prognosis can thus be the first step of a diagnosis of an autoimmune disease, such as MS. For example, the methods for early prognosis provided herein, may be an initial step of starting a diagnosis, e.g. deciding if the further diagnostics examinations are to be performed / necessary according to the prognosed risk. This is advantageous since diagnosis of e.g. MS is a multistep, lengthy diagnosis performed e.g. according to the McDonald criteria7Diagnosis of MS is described in the section “Methods” above in detail. Thus, prognosis as defined in the methods for early prognosis provided herein may be part of a diagnostic method.

[0429] Accordingly, the invention provides a method of diagnosis of an autoimmune disease, preferably MS wherein the method comprises:

[0430] (a) contacting a biological sample of a subject with the peptide, or the composition, or the antigen of the invention,

[0431] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0432] The invention also provides the peptide, or the composition, or the antigen of the invention, for use in a method of diagnosis of an autoimmune disease, preferably multiple sclerosis (MS). The invention also provides the peptide, or the composition, or the antigen of the invention, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably multiple sclerosis (MS). The invention also provides the peptide, or the composition, or the antigen of the invention, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably MS wherein the method comprises:

[0433] (a) contacting a biological sample of a subject with the peptide, or the composition, or the antigen of the invention,

[0434] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

[0435] Since the prognosis can be performed as a first step of diagnosis, the method of diagnosis or the diagnostic use provided herein may comprise any method steps of the methods for early prognosis provided herein. Accordingly, all features described in relation to the methods for early prognosis in the section “Methods” above, apply mutatis mutandis to the method of diagnosis or the diagnostic use of the present invention. The method of diagnosis or the diagnostic use may be in vitro or ex vivo. In one aspect, the method of diagnosis or the diagnostic use do not comprise a step of obtaining a biological sample of a subject. In another aspect, the method of diagnosis or the diagnostic use can further comprise a step of obtaining a biological sample of a subject.

[0436] The features described herein in context of the diagnostic use of the peptide, or the composition, or the antigen of the invention apply mutatis mutandis to the method of diagnosis provided herein. As explained above in context of the methods for early prognosis provided herein, a diagnosis can be followed up by early treatment. Accordingly, the invention provides a peptide, or a composition, or an antigen of the invention, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably MS wherein the method comprises:

[0437] (a) contacting a biological sample of a subject with the peptide, or the composition, or the antigen of the invention,

[0438] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen, wherein the method further comprises advising early treatment if the subject is at risk of contracting the autoimmune disease.

[0439] In one embodiment, the step of advising early treatment comprises advising treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0- 1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0440] In some embodiments, the step of advising early treatment comprises advising disease-modifying treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon 0- 1 a (Avonex, Rebif), Interferon 0-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0441] In some embodiments, the step of advising early treatment comprises advising symptomatic treatment with a drug selected from the list consisting of Alprostadil (Caveiject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0442] In another embodiment, the treatment comprises administering Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon P-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), or Vardenafil (Levitra, Staxyn, and others) and the like, or any combinations thereof to the subject.

[0443] In some embodiments, the treatment is a disease-modifying treatment and comprises administering Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon P-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0444] In some embodiments, the treatment is a symptomatic treatment and comprises administering Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0445] In one aspect of the invention, the diagnostic method and / or the diagnostic use provided herein do not comprise a treatment step.

[0446] In another aspect of the invention, the diagnostic method and / or the diagnostic use provided herein may comprise a further treatment step as defined above in the context of advising early treatment. Accordingly, the invention provides a method of diagnosis of an autoimmune disease, preferably MS wherein the method comprises: (a) contacting a biological sample of a subject with the peptide, or the composition, or the antigen of the invention,

[0447] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen, wherein the method further comprises treatment of the autoimmune disease.

[0448] The invention also provides the peptide, or the composition, or the antigen of the invention, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably MS wherein the method comprises:

[0449] (a) contacting a biological sample of a subject with the peptide, or the composition, or the antigen of the invention,

[0450] (b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen, wherein the method further comprises treatment of the autoimmune disease.

[0451] In one embodiment, the step of treatment of the autoimmune disease comprises treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon P-lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0452] In some embodiments, the step of treatment of the autoimmune disease comprises a diseasemodifying treatment with a drug selected from the list consisting of Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon -la (Avonex, Rebif), Interferon -lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0453] In some embodiments, the step of treatment of the autoimmune disease comprises symptomatic treatment with a drug selected from the list consisting of Alprostadil (Caveiject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0454] In another embodiment, the treatment of the autoimmune disease comprises administering Alemtuzumab (Lemtrada, Campath), Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Cladribine (Mavenclad), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon - lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Oxybutynin (Anturol, Ditropan, and others), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Sildenafil (Revatio, Viagra), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), Ublituximab (Briumvi), or Vardenafil (Levitra, Staxyn, and others) and the like, or any combinations thereof to the subject. In some embodiments, the treatment of the autoimmune disease is a disease-modifying treatment and comprises administering Alemtuzumab (Lemtrada, Campath), Cladribine (Mavenclad), Dimethyl fumarate / Diroximel fumarate (Tecfidera / Vumerity and generics), Monomethyl fumarate (BAFIERTAM), Fingolimod (Gilenya and biosimilars), Glatiramer acetate (Copaxone and follow-on drugs), Interferon P-1 a (Avonex, Rebif), Interferon -lb (Betaseron, Extavia), Natalizumab (Tysabri and biosimilars), Ocrelizumab (Ocrevus), Ofatumumab (Kesimpta), Ozanimod (Zeposia), Ponesimod (Ponvory), Rituximab (Rituxan, Mab Thera, Truxima), Siponimod (Mayzent), Teriflunomide (Aubagio and generics), or Ublituximab (Briumvi) and the like or any combinations thereof.

[0455] In some embodiments, the treatment of the autoimmune disease is a symptomatic treatment and comprises administering Alprostadil (Caverject, Edex, and others), Baclofen (Gablofen, Lioresal, and others), Bupropion (Wellbutrin, Aplenzin, and others), Clonazepam (Klonopin and others), Cyclophosphamide (Cytoxan and others), Daclizumab (Zenapax), Dantrolene (Dantrium and others), Dexamethasone (Maxidex, Ozurdex, and others), Diazepam (Valium, Diastat, and others), Oxybutynin (Anturol, Ditropan, and others), Sildenafil (Revatio, Viagra), Tizanidine (Zanaflex and others), Tolterodine (Detrol), Trospium (Sanctura and others), or Vardenafil (Levitra, Staxyn, and others) and the like or any combinations thereof.

[0456] Uses

[0457] The peptide and / or composition of the present invention may be used as an antigen, e.g. for detecting and / or quantifying the binding of antibodies to said peptide and / or composition. The peptide and / or composition of the present invention may for example be used in any of the detection / quantification assays provided herein, such as Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC), Flow Cytometry and / or Fluorescence- Activated Cell Sorting (FACS) and the like. For the use as an antigen, the peptide and / or composition of the invention may be coated onto the surface of a membrane, microtiter plate, bead, gel, or column or is immobilized in a gel. For the use as an antigen, the peptide and / or composition may preferably be coated onto the surface of a microtiter plate to be used in an ELISA.

[0458] Kits

[0459] The invention also provides kits for carrying out the methods of the invention. The kit of the invention can comprise the peptide, composition and / or antigen of the invention and means for detecting and / or quantifying antibodies binding to said peptide, composition, or antigen. The kit may further comprise instructions for use. The kit may further comprise an anti-human Immunoglobulin G (IgG) antibody. Such an antibody can be used to detect and / or quantify binding of EBNA381-452 derived antibodies to the peptide, composition or antigen of the invention in a biological sample of a subject. Antibodies binding to the peptide, composition, or antigen comprised in the kit of the present invention are preferably detected and / or quantified by ELISA. Accordingly, the means for detecting and / or quantifying antibodies binding to the peptide, composition, or antigen of the invention comprised in the kit of the invention are preferably means to perform an Enzyme-Linked Immunosorbent Assay (ELISA).

[0460] In one aspect of the invention, the peptide, composition, and / or antigen of the invention does not comprise the amino acid sequence SSSSGSPPRRPPPGR (SEQ ID NO: 58).

[0461] In one aspect of the invention, the peptide, composition, and / or antigen of the invention does not comprise the amino acid sequence PPPGRRP (SEQ ID NO: 59).

[0462] In one aspect of the invention, the peptide, composition, and / or antigen of the invention does not comprise the amino acid sequence PDVPPGAI (SEQ ID NO: 60).

[0463] In one aspect of the invention, the peptide, composition, and / or antigen of the invention does not comprise the amino acid sequence PGAIEQGPA (SEQ ID NO: 61).

[0464] In one aspect of the invention, the peptide, composition, and / or antigen of the invention does not comprise the amino acid sequence GPSTGPRG (SEQ ID NO: 62).

[0465] In one aspect of the invention, the peptide, composition, and / or antigen of the invention does not comprise the amino acid sequence of any one of SSSSGSPPRRPPPGR (SEQ ID NO: 58), PPPGRRP (SEQ ID NO: 59), PDVPPGAI (SEQ ID NO: 60), PGAIEQGPA (SEQ ID NO: 61), and / or GPSTGPRG (SEQ ID NO: 62).

[0466] Unless otherwise defined, all terms of art, notations and other scientific terminology used herein are intended to have the meanings commonly understood by those of skill in the art to which this invention pertains. In some cases, terms with commonly understood meanings are defined herein for clarity and / or for ready reference, and the inclusion of such definitions herein should not necessarily be construed to represent a difference over what is generally understood in the art. The techniques and procedures described or referenced herein are generally well understood and commonly employed using conventional methodologies by those skilled in the art. As appropriate, procedures involving the use of commercially available kits and reagents are generally carried out in accordance with manufacturer-defined protocols and conditions unless otherwise noted.

[0467] As used herein, the singular forms “a,” “an,” and “the” include the plural referents unless the context clearly indicates otherwise. The terms “include,” “such as,” and the like are intended to convey inclusion without limitation, unless otherwise specifically indicated.

[0468] As used herein, the term “or” is generally employed in its usual sense including “and / or” unless the content clearly dictates otherwise. The term “and / or” means one or all of the listed elements or a combination of any two or more of the listed elements.

[0469] As used herein, the term “comprising” also specifically includes embodiments “consisting of’ and “consisting essentially of’ the recited elements, unless specifically indicated otherwise.

[0470] As used herein, the term “about” indicates and encompasses an indicated value and a range above and below that value. In certain embodiments, the term “about” indicates the designated value ± 10%, ± 5%, or± 1%. In certain embodiments, where applicable, the term “about” indicates the designated value(s) ± one standard deviation of that value(s).

[0471] In some embodiments any one of the methods or uses described herein can be in vitro and / or ex vivo or in vivo. In some embodiments any of the method steps described herein can be in vitro and / or ex vivo or in vivo.

[0472] The disclosures in context of the methods described herein are disclosed as corresponding uses mutatis mutandis. The disclosures in context of the uses described herein are disclosed as corresponding methods mutatis mutandis. In particular, the disclosures in context of the diagnostic use provided herein are disclosed as the corresponding diagnostic method mutatis mutandis.

[0473] In one aspect, the methods of the present invention are not methods for treatment of the human or animal body by therapy. In a further aspect, the methods of the present invention are not processes for modifying the germ line genetic identity of human beings. In one aspect, the methods of the present invention are in vitro or ex vivo methods. In a further aspect, the methods of the present invention are non-therapeutic or non-medical. In a further aspect, the methods of the present invention are not processes for modifying the genetic identity of animals which are likely to cause them suffering without any substantial medical benefit to man or animal.

[0474] Brief description of the drawings:

[0475] Figure 1 and Figure 1.1: Individual EBNA38i-452-derived IgG antibody titers were assessed in N=270 MS patients and N=270 controls at the time point of MS diagnosis or at a matched time point for healthy controls (± 1 month). The dashed black line indicates the detection limit of EBNA38i-452-specific IgG antibody titers. Antibody titers were compared between the groups using the Mann-Whitney-Test. Statistical significance was set at p < 0.05 and assessed using GraphPad Prism 10. MS: Multiple Sclerosis, OD: optical density.

[0476] Figure 2: EBNA38i-452-specific IgG antibody titer of N=270 MS patients and N=270 controls were assessed at three different time points: EBNA38i-452-specific IgG antibody titers were assessed for MS patients at the time point of MS diagnosis as well as between 1-2 years or 3-4 years before the MS diagnosis, respectively. For healthy controls, retrospective follow-up serum samples were matched to the time point of the MS diagnosis (± 1 month). Each dot represents one individual MS patient or healthy control. The red line indicates the respective median. The dashed black line indicates the detection limit of EBNA38i-452-specific IgG antibody titers. Antibody titers were compared between the groups using the Mann-Whitney-Test. Statistical significance was set at p < 0.05 and assessed using GraphPad Prism 10. MS: Multiple Sclerosis, OD: optical density.

[0477] Figure 3 and Figure 3.1: Accumulation EBNA38i-452-mediated immune responses is associated with MS. (A-D) EBNA-specific IgG antibody responses were assessed in N=270 MS patients and N=270 healthy controls using indicated (A) EBNA38i-452-derived peptide pools or (B) individual EBNA386-405-, EBNA393-412-, EBNA409-428-, EBNA426-445- or corresponding CNS-derived GlialCAM37o-389-, CRYAB2-21-, MBP205-224-, ANO2135-154 peptides. The dashed black line indicates the cut-off for detection. (C, D) High-level EBNA-specific or CNS-specific antibody levels were first defined by ROC analysis (Fig.4A) and then compared between MS patients and controls. (E) Distribution of high-level EBNA-specific or CNS-specific CD19+B cell (Fig.3B), CD4+T cell (Fig.3C), and CD8+T cell (Fig.3D) levels was defined by ROC analysis and then compared between N=20 MS patients, N=31 healthy controls with >3 EBNA-derived antibody responses (HC cAB-3) and N=49 healthy controls with <2 EBNA-derived antibody responses (HC cAB-2). Statistical differences were assessed by (A) Kruskal-Wallis and Dunn’s multiple comparison test, (B) Mann-Whitney test, (C-E) %2-Test. ABs: Antibodies, CNS: Central Nervous System, HC: Healthy Control, MS: Multiple Sclerosis.

[0478] Figure 4: (A) ROC analysis to define high-level EBNA386-405- (cut-off: < 59.5%), EBNA393-412- (cut-off: < 61.97%), EBNA409-428- (cut-off: < 57.04%), and EBNA426-445- (cut-off: < 51.15%) antibody titers as well as corresponding high-level GlialCAM37o-389- (cut-off: < 46.74%), CRYAB2-21- (cut-off: < 33.08%), MBP205-224- (cut-off: < 52.99%), ANO2i35-i54- (cut-off: < 56.89%) antibody titers. Antibody levels were compared between N=270 MS patients and N=270 healthy controls. (B) ROC analysis to define high-level EBNA386-405- (cut-off: < 0.001583%), EBNA393-412- (cut-off: < 0.0003797%), EBNA409-428- (cut-off: < 0.001806%), and EBNA426-445- (cut-off: < 0.001815%) CD19+B cell frequencies as well as corresponding high-level GlialCAM37o-389- (cut-off: < 0.001064%), CRYAB2-21- (cut-off: < 0.0003797%), MBP205-224- (cutoff: < 0.001806%), ANO2135-154- (cut-off: < 0.001815%) -specific CD19+B cell frequencies. CD19+B cell frequencies were compared between N=20 MS patients and N=80 healthy controls. (C) ROC analysis to define high-level EBNA386-405- (cut-off: < 0.3250), EBNA393-4i2-(cut-off < 0.2998%), EBNA409-428- (cut-off: < 0.7113%), andEBNA426-445- (cut-off: < 0.1777%) CD4+T cell frequencies as well as corresponding high-level GlialCA 37o-389- (cut-off: < 0.1340%), CRYAB2- 21- (cut-off: < 0.1256%), MBP205-224- (cut-off: < 0.1223%), ANO2i35-i54- (cut-off: < 0.1165%) - specific CD4+T cell frequencies. CD4+T cell frequencies were compared between N=20 MS patients and N=80 healthy controls. (D) high-level EBNA386-405- (cut-off: < 0.01128), EBNA393- 412- (cut-off: < 0.01133%), EBNA409-428- (cut-off: < 0.008129%), and EBNA426-445- (cut-off: < 0.004754%) specific CD8+T cell frequencies as well as corresponding high-level GlialCAM37o- 389- (cut-off: < 0.004573%), CRYAB2-21- (cut-off: < 0.002575), MBP205-224- (cut-off: < 0.002522%), ANO2135-154- (cut-off: < 0.002894%) -specific CD8+T cell frequencies. CD8+T cell frequencies were compared between N=20 MS patients and N=80 healthy controls. AUC: Area under the Curve.

[0479] Figure 5 and Figure 5.1: (A) EBNA-specific or CNS-specific CD4+T cells, CD8+T cells, and CD19+B cell levels were assessed in N=20 MS patients, N=31 healthy controls with >3 EBNA- derived antibody responses (HC cAB-3) and N=49 healthy controls with <2 EBNA-derived antibody responses (HC cAB-2). Heatmap shows the individual coefficients of determination (r2) from non-linear regression between EBNA-derived and CNS-derived antibody titers and respective EBNA-derived and CNS-derived CD19+B cell, CD4+T cell, and CD8+T cell levels. (B-D) EBNA386-405-, EBNA393-412-, EBNA409-428-, EBNA426-445- or corresponding CNS-derived GlialCAM37o-389-, CRYAB2-21-, MBP205-224-, ANO2135-154- peptides-specific (B) CD19+B cell, (C) CD4+T cell, or (D) CD8+T cell levels were assessed in N=20 MS patients, N=31 healthy controls with >3 EBNA-derived antibody responses (HC cAB-3) and N=49 healthy controls with <2 EBNA-derived antibody responses (HC cAB-2) by flow-cytometry. (B-D) Statistical differences were assessed between the groups by Kruskal-Wallis and Dunn’s multiple comparison test. ABs: Antibodies, HC: Healthy Control, MS: Multiple Sclerosis.

[0480] Figure 6 and Figure 6.1: Impaired control of EBNA38i-452-mediated immune responses is associated with MS. (A) Distribution of NKG2C+NKG2D' NK cell, NKG2C NKG2D+NK cell, NKG2C+NKG2D+NK cell, and NKG2C NKG2D' NK cell subsets were assessed in N=20 MS patients, N=31 healthy controls with >3 EBNA-derived antibody responses (HC cAB-3) and N=49 healthy controls with <2 EBNA-derived antibody responses (HC cAB-2) by flow-cytometry. (B) Distribution of the combination of the NKG2DHNK / LNKvariants, HCMV-serostatus, and KLRC2wt / delvariants in N=270 MS patients, N=31 HC cAB-3, and N=239 HC cAB-2. (C) Control of EBNA- or CNS-specific CD4+T cells, CD8+T cells, and CD19+B cells by CD56+NK cells. Data are shown as the ratio of the lysis of EBNA-specific or CNS-specific to respective nonspecific controls. (D) CD56+NK cell subsets after expansion with IL-12, IL-15 and IL-18 or noncytokine controls. Data are shown as the distribution and fold change (to pre-expansion, Fig.6A) of indicated NK cell subsets. (E) Percentage of EBV-infected (EBER+) CD19+B cells in N=20 MS patients, N=31 HC cAB-3, and N=49 HC cAB-2. (F) Distribution of LMP-1 peptide variants in EBV-isolates of N=270 MS patients, N=31 HC cAB-3, and N=239 HC cAB-2. The interpretation of the LMP-1 peptide variants in highly ( TT), intermediate (Tf) and low (T) NKG2A-inhibiting variants is based on6. Statistical differences were assessed by (A, D) Two-Way ANOVA and Dunn’s multiple comparison test, (C, E) Kruskal -Wallis and Dunn’s multiple comparison test, (B, F) %2-Test and Fisher’ s-exact test. ABs: Antibodies, Del: Deletion, HC: Healthy Control, LDH: Lactate Dehydrogenase, MS: Multiple Sclerosis, Neg.: Negative, Pos.: Positive, Wt: Wild-Type.

[0481] Figure 7 and Figure 7.1 A-D: (A) Frequencies of EBNA386-405-, EBNA393-412-, EBNA409-428-, EBNA426-445- or corresponding CNS-derived GlialCAM3?o-389-, CRYAB2-21-, MBP205-224-, ANO2135-154- peptides-specific CD19+B cells, CD4+T cells, or CD8+T cells were assessed in N=20 MS patients, N=31 healthy controls with >3 EBNA-derived antibody responses (HC cAB-3) and N=49 healthy controls with <2 EBNA-derived antibody responses (HC cAB-2) by flowcytometry before killing assays. Statistical differences were assessed by Two-Way ANOVA and Dunn’s multiple comparison test. (B-E) Control of EBNA- or CNS-specific CD4+T cells, CD8+T cells, and CD19+B cells by CD56+NK-cells after stimulation with IL-12, IL-15 and IL-18 or non-cytokine controls. Data are shown as (B, D) the ratio of the lysis of EBNA- or CNS-specific to respective non-specific controls or (C, E) the ratio of the lysis of EBNA- or CNS-specific cells before and after expansion with (C) IL-12, IL-15 and IL-18 or (E) respective non-cytokine controls. The control of EBNA- or CNS-specific CD4+T cells, CD8+T cells, and CD19+B cells was assessed in N=20 MS patients, N=31 healthy controls with >3 EBNA-derived antibody responses (HC cAB-3) and N=49 healthy controls with <2 EBNA-derived antibody responses (HC cAB-2) by LDH release assay. Statistical differences were assessed by Kruskal -wallis and Dunn’s multiple comparison test. ABs: Antibodies, HC: Healthy Control, LDH: Lactate Dehydrogenase, MS: Multiple Sclerosis.

[0482] Figure 8: Evaluation of EBNA386-405 and GlialCAM37o-389-specific immune responses.

[0483] (A-D) Evaluation of EBNA386-405 IgG antibody responses. EBNA386-405-specific IgG antibody titers were assessed in N=270 controls by ELISA. The ELISA titers were normalized to one control plasma. EBNA386-405 IgG antibody titers were then compared according to the (A) sex, (B) age,

[0484] (C) total EBNA- 1 -specific IgG antibody titers, and (D) HLA-DRB1 genotypes of the controls. The groups were compared using (A) the Mann-Whitney-Test, (B-C) simple linear regression and

[0485] (D) Kruskal -Wallis and Dunn’s multiple comparison test. Each dot represents one individual patient. The (red) line represents (A,D) the corresponding means or (B,C) the linear regression. (E-G) Evaluation of GlialCAM37o-389-specific B-cell, plasma cell and IgG antibody responses. GlialCAM37o-389-specific (E) CD19+B-cell or (F) CD138+plasma cells were assessed in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols by flow cytometry. Data are shown as the percentage of CFSElowGlialCAM37o-389-specific (E) CD19+B-cell or (F) CD138+plasma cells, in comparison to all (E) CD19+B-cell or (F) CD138+plasma cells, respectively. (G) GlialCAM37o- 389 IgG antibody titers were assessed in N=270 MS patients, N=162 EBNAlow, and N=108 EBNAhlghcontrols by ELISA. The ELISA titers were normalized to one control plasma. The dashed black line indicates the 46% normalized IgG titer (100% negative predictive value) to discriminate between EBNAlowand EBNAhlghcontrols. (E-G) The groups were compared by Kruskal -Wallis and Dunn’s multiple comparison test. Each dot represents one individual patient. The (red) line represents the corresponding means. (H) Gating strategy for the identification of EBNA386-405-specific IgG antibody -mediated ADCP and ADCC responses. CD14+CD16+monocytes from 12 healthy EBV-seropositive blood donors were differentiated into CD14+CD16+Ml macrophages and then stimulated with AF647 NHS Ester-linked GlialCAM37o-389 peptides in the presence or absence of individual plasma of N=270 MS patients, N=162 EBNAlow, and N=108 EBNAhlghcontrols. GlialCAM37o -389-specific ADCP responses were assessed as the median AF647 NHS Ester MFI of CD14+CD16+Ml macrophages. GlialCAM37o-389-specific ADCC responses were assessed in parallel as CD107a+CD14+CD16+Ml macrophages. One representative example of two parallel experiments, in the presence or absence of plasma of one EBNAlowcontrol individual is shown. (I) Gating strategy for the identification of EBNA386-405-specific CD8+T-cells (panels in first row), CD4+T-cells (panels in second row) and CD138+plasma cells (panels in third row). PBMCs from one control individual were cultured in the presence of the EBNA386-405 peptide. EBNA386-405-specific CD8+T-cells, CD4+T-cells and CD138+plasma cells were identified as CFSElowcells (blue). (J) CD4+T-cells from one control individual were cultured in the presence or absence of the EBNA386-405 peptide. EBNA386-405-specific CD4+T-cells were identified as CFSElowcells (blue / light grey) in comparison to the non-peptide control (red / dark grey). One representative example is shown. ADCC: antibody-dependent cellular cytotoxicity, ADCP: antibody-dependent cellular phagocytosis, DU: DRG units, MFI: mean fluorescence intensity, MS: Multiple Sclerosis, ns: not significant.

[0486] Figure 9: Evaluation of EBNA386-405- and GlialCAM37o-389-specific immune responses.

[0487] (A) EBNA386-405-specific IgG antibodies titers were assessed in N=270 MS patients, N=162 EBNAlow, and N=108 EBNAhlghcontrols by ELISA. The ELISA titers were normalized to one control plasma. The dashed black line indicates the 46% normalized IgG titer (100% negative predictive value) to discriminate between EBNAlowand EBNAhlghcontrols. (B-C) Ml macrophages-mediated and GlialCAM37o-389-specific (B) ADCP and (C) ADCC responses from N=270 MS patients, N=162 EBNAlow, and N=108 EBNAhlghcontrols were assessed using monocyte-derived Ml macrophages from 12 healthy EBV-seropositive blood donors. (B) GlialCAM37o -389-specific ADCP was assessed as AF647 MFI by flow cytometry. (C) GlialCAM37o -389-specific ADCC was assessed as CD107a+CD14+cells by flow cytometry. (B-C) The data were normalized to one control plasma. (D-F) EBNA386-405-specific (D) CD138+plasma cells, (E) CD4+T-cells, and (F) CD8+T-cells were assessed in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols by flow cytometry. Data are shown as the percentage of CFSElow EBNA386-405-specific (D) CD138+plasma cells, (E) CD4+T-cells, and (F) CD8+T-cells in comparison to total (D) CD138+plasma cells, (E) CD4+T-cells and (F) CD8+T-cell levels. (G-J) GlialCAM37o-389-specific (G, I) CD4+T-cells and (H, J) CD8+T-cells were assessed in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols by flow cytometry. Data are shown as a percentage of CFSElowGlialCA 37o-389-specific (G, I) CD4+T-cells and (H, J) CD8+T-cells in comparison to (G,I) total or (H,J) EBNA386-405-specific CD4+T-cells and CD8+T-cell levels, respectively. (A-J). The groups were compared by Kruskal -Wallis and Dunn’s multiple comparison test. Each dot represents one individual patient. The (red) line represents the corresponding means. ADCC: antibody-dependent cellular cytotoxicity, ADCP: antibodydependent cellular phagocytosis, MFI: mean fluorescence intensity, MS: Multiple Sclerosis, ns: not significant.

[0488] Figure lOand Figure 10.1 A-B and F-I: Control of GlialCAM37o-389-specific immune responses.

[0489] (A-B) Phenotype of GlialCAM37o-389-specific cells after stimulation with the GlialCAM37o-389 peptide. Total PBMCs from 24 healthy control individuals were stimulated with the GlialCAM37o- 389 peptide or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC) for 14 days. GlialCAM37o-389-specific (GlialCAM37o-389) or total (NC, NPC) CD8+T-cells, CD4+T-cells or CD138+plasma cells were then sorted, analyzed for the expression of PD-1 by cytometry and pooled for the cell killing experiments. (A) PD1 expression of GlialCAM37o-389-specific (GlialCAM37o-389) or total (NC, NPC) CD8+T-cells, CD4+T-cells or CD138+plasma cells. Data are shown as the mean cellular fractions after pooling of 24 independent experiments ± SEM. The individual panels in the bars relate from top to bottom to: PD1 CD8+T cells, PD1+CD8+T cells, PD1 CD4+T cells, PD1+CD4+T cells, PD1 CD138+plasma cells and PD1+CD138+T plasma cells. (B) Characterization of GlialCAM37o-389-specific CD8+T- cells. GlialCAM37o-389-specific (GlialCAM37o-389) or total (NC, NPC) CD8+T-cells were analyzed for the expression of LAG3, PD1, TGIT, TIM3, and NKG2A. Each fraction represent the mean frequency of the LAG3 PD1+ / TGIT+ / TIM3 NKG2A', LAG3+PD1+TGIT+TIM3 NKG2A' and LAG3+PD1+TGIT+TIM3+NKG2A+CD8+T cell subsets (±SEM). The individual panels in the bars relate from top to bottom to: LAG3+PD1+TGIT+TIM3+NKG2A+, LAG3+PD1+TGIT+TIM3’ NKG2A' and LAG3 PDrTGIT TIM3 NKG2A'. (A-B) Groups were compared using the Two- Way-ANOVA. (C-D) Control of GlialCAM37o-389-specific immune responses by CD8+T-cells. PBMCs from 24 healthy control individuals were first stimulated with the GlialCAM37o-389-peptide or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non- peptide control, NPC) and then co-cultured with enriched CD8+T-cells from the same donor. The CD8+T-cell activation level was assessed either by (C) IL-2 ELISA or by (D) IFNy ELISA. (C- D) The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. Each dot represents one individual sample from one healthy control. (E) Gating strategy for the identification of the NKG2D+NK cell-mediated cytotoxicity against GlialCAM37o-389-specific cells. PBMCs from one healthy blood donor were first stimulated with the GlialCAM37o-389-peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). GlialCAM37o-389-specific (GlialCAM37o-389) or total (NPC, NC) CD8+T-cells, CD4+T-cells or CD138+plasma cells were then sorted and co-cultured with sorted NKG2D+NK cell from the same donor. Cytotoxic NKG2D+NK cell responses were assessed as CD107a+CD56+NKG2D+NK cells in response to GlialCAM37o-389-specific B-cells and T-cells (blue, 12.4% CD107a+CD56+NKG2D+NK cells), the NC (red 2.2% CD107a+CD56+NKG2D+NK cells), or the NPC (orange 2.5% CD107a+CD56+NKG2D+NK cells). One representative example is shown. (F-G) NKG2C+NK cell responses: (F) Simple linear regression of EBNA386-405-specific IgG antibody titers and NKG2C+NK cell frequencies in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols. Each dot represents one individual sample. (G) PBMCs from 24 healthy control individuals (KLAC2wt / wt: N=12 or KLRC2wt / del: N=12) were first stimulated with the GlialCAM37o-389-peptide or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). GlialCAM37o-389- specific (GlialCAM37o-389) or total (NPC, NC) CD8+T-cells, CD4+T-cells or CD138+plasma cells were then sorted and co-cultured with sorted NKG2C+NK cell from the same donor. The LDH release was then assessed in the supernatant. Results are shown as fold change of the GlialCAM37o- 389-specific cells or the NC, normalized to the NPC. The dashed black line indicates an equal activation of NK cells against GlialCAM37o-389-specific and non-GlialCAM37o-389-specific cells. The groups were compared by the Wilcoxon matched T-test. (H-I) NKG2D+NK cell responses: (H) Simple linear regression of EBNA386-405-specific IgG antibody titers and NKG2D+NK cell frequencies in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols. Each dot represents one individual sample. (I) PBMCs from 24 healthy control individuals (NKG2DHNK / HNK: N=8, NKG2DHNK / LNK: N=8, or NKG2DLNK / LNK: N=8) were first stimulated with the GlialCAM37o -389-peptide or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). GlialCAM37o-389-specific (GlialCAM37o-389) or total (NPC, NC) CD8+T-cells, CD4+T-cells or CD138+plasma cells were then sorted and co- cultured with sorted NKG2D+NK cell from the same donor. The LDH release was then assessed in the supernatant. Results are shown as fold change of the GlialCAM37o-389-specific cells or the NC, normalized to the NPC. The dashed black line indicated an equal activation of NK cells against GlialCAM37o-389-specific and non-GlialCAM37o-389-specific cells. The groups were compared by the Wilcoxon matched T-test. LDH: lactate dehydrogenase, MFI: mean fluorescence intensity, MS: Multiple Sclerosis, NC: negative control, NPC: non-peptide control, ns: not significant, wt: wild type.

[0490] Figure 11 and Figure 11.1 G and J: Control of GlialCAM37o-389-specific immune responses.

[0491] (A-D) PBMCs from 24 healthy control individuals were first stimulated with the GlialCAM37o-389- peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (none-peptide control, NPC). Sorted GlialCAM37o-389-specific (GlialCAM37o- 389) or total (NC, NPC) CD4+T-cells, CD8+T-cells, and CD138+plasma cells were then pooled and co-cultured with (A) Jurkat T-cells, (B) THP-1 monocytes, or (C) NK92 NK cells. The cell activation level was either assessed by (A) IL-2 ELISA, (B) flow cytometry (AF647 MFI), or (C) IFNy ELISA. (A-C) The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. (D) GlialCAM37o-389-specific CD4+T-cells, CD8+T-cells, and CD138+plasma cells (GlialCAM37o-389) or total CD4+T-cells, CD8+T-cells and CD138+plasma cells (NC) from 24 healthy blood donors were co-cultured with sorted KIR2DS1+, KIR3DS1+, NKp46+(NCR1), NKp44+(NCR2), NKp30+(NCR3), NKG2D+, orNKG2C+CD56+NK cell subsets from the same donor. The percentage of activated and cytotoxic (CD107a+) KIR2DS1+, KIR3DS1+, NKp46+(NCR1), NKp44+(NCR2), NKp30+(NCR3), NKG2D+or NKG2C+CD56+NK cell subsets was assessed by flow cytometry. Results are shown as fold change of the GlialCAM37o-389-specific (GlialCAM37o-389) to total cells, stimulated without peptides (NC). The dashed black line indicated an equal activation of NK cells against GlialCAM37o-389-specific cell (GlialCAM37o-389) and cells, stimulated without peptides (NC). Each dot represents one individual sample from one healthy control. The groups were compared by Wilcoxon matched T-test. (E-G) NKG2C+NK cell responses. (E) NKG2C+NK cell frequencies were assessed in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols by flow cytometry and compared by the Kruskal-Wallis and the Dunn’s multiple comparison test. (F) GlialCAM37o-389-specific CD4+T-cells, CD8+T-cells and CD138+plasma cells (GlialCAM37o-389) or total CD4+T-cells, CD8+T-cells and CD138+plasma cells (NC) from 24 healthy blood donors were co-cultured with sorted NKG2C+CD56+NK cells from the same donor. The controls expressed either the V / . / C2"l"l(N= l 2), or V / . / C2"l dcl(N= l 2) genotype. The percentage of activated and cytotoxic (CD107a+) NKG2C+CD56+NK cells was assessed by flow cytometry. Results are shown as fold change of the GlialCAM37o-389-specific (GlialCAM37o-389) to non-specific cells (NC). The dashed black line indicated an equal activation of NK cells against GlialCAM37o-389-specific (GlialCAM37o-389) and non-specific cells (NC). The groups were compared by the Wilcoxon matched T-test. (G) Distribution of the HCMV-serostatus, KLRC2 variants, and HCMV-encoded UL40 variants of the infecting HCMV-variant. Fractions represent the relative frequency of each combination of the HCMV-serostatus, KLRC2 variants, and HCMV-encoded UL40 variants. UL40 variants were categorized into high ( )_and low (T)_binders as shown in Table 11. The frequency of the combination was compared between the cohorts by the Chi2Test. The individual panels in the bars relate from left to right to: HCMV neg.; HCMV pos. + KLRC2del / del; HCMV pos. + KLRC2wt / del+ U140T; HCMV pos. + KLRC2wt / wt+ U140T; HCMV pos. + KLRC2wt / del+ U140TT; and HCMV pos. + KLRC2wt / wt+ U140TT- (H-J) NKG2D+NK cell responses. (H) NKG2D+NK cell frequencies were assessed in N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols by flow cytometry and compared by the Kruskal-Wallis and the Dunn’s multiple comparison test. (I) GlialCAM37o-389-specific CD4+T- cells, CD8+T-cells and CD138+plasma cells (GlialCAM37o-389) or non-GlialCAM37o-389-specific CD4+T-cells, CD8+T-cells and CD138+plasma cells (NC) from 24 healthy blood donors were co-cultured with sorted NKG2D+CD 56+NK cells from the same donor. The healthy blood donors expressed either the NKG2DHNK / HNK(N=8), NKG2DHNK / LNK(N=8) or NKG2DLNK / LNK(N=8) genotype. The percentage of activated and cytotoxic (CD107a+) NKG2D+CD56+NK cells was assessed by flow cytometry. Results are shown as fold change of the GlialCAM37o-389-specific to non-specific cells. The dashed black line indicated an equal activation of NK cells against GlialCAM37o-389-specific (GlialCAM37o-389) and non-GlialCAM37o-389-specific (NC) cells. The groups were compared by the Wilcoxon matched T-test. (J) Distribution of NKG2DHNK / LNKvariants in N=270 MS patients, N=162 EBNAlow, and N=108 EBNAhlghcontrols. Fractions represent the relative frequency of NKG2DLNK / LNK, NKG2DHNK / LNKand NKG2DHNK / HNKvariants. The distribution of the variants was compared between the cohorts by the Chi2Test. The individual panels in the bars relate from right to left to: NKG2DLNK / LNK, NKG2DHNK / LNKand NKG2DHNK / HNK. Del: deletion, HCMV: Human Cytomegalovirus, HNK: high NK cytotoxicity, LNK: low NK cytotoxicity, MS: Multiple Sclerosis, NC: negative control, NPC: non-peptide control, ns: not significant, wt: wild type.

[0492] Figure 12: GlialCAM37o-389-specific immune cells from MS patients upregulate HLA-E via the EBV-mediated secretion of IL-27.

[0493] (A-C) HLA-E expression of GlialCAM37o-389-specific (A) CD4+T-cells, (B) CD8+T-cells, and (C) CD138+plasma cells, derived from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols. The HLA-E expression was assessed after PBMC stimulation with the GlialCAM37o-389 peptide via flow cytometry. (D-F) NKG2A inhibition assay. Sorted GlialCAM37o-389-specific (D) CD4+T-cells, (E) CD8+T-cells and (F) CD138+plasma cells, derived from N=20 MS patients, N=48 EBNAlowand N=32 EBNAhlghcontrols were co-cultured with sorted NKG2A+NKG2C' NKG2D’ NK cells from 24 healthy blood donors. The NKG2A+NKG2C NKG2D' NK cell activation level was assessed by IFNy ELISA. Data are shown as the mean from all 24 independent experiments, reflecting 24 healthy blood donors. (G-I) EBV reactivation in GlialCAM37o-389- specific immune cells. PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols were stimulated with the GlialCAM37o-389 peptide. (G) The EBV reactivation was then assessed by EBV qPCR of the supernatant. The dashed black line indicates the detection limit of the EBV qPCR. (H-I) The number of EBV-infected (EBV EBER+) (H) total or (I) GlialCAM37o- 389-specific B-cells was afterward assessed by flow cytometry. Data are shown as EBER+per IxlO7B-cells. (J) IL-27 secretion of EBV-infected GlialCAM37o-389-specific cells. PBMCs from N=20 MS patients, N=48 EBNAlowand N=32 EBNAhlghcontrols were stimulated with the GlialCAM37o- 389 peptide and the IL-27 concentration in the supernatant was assessed by IL-27 ELISA. (K-L) Supernatant transfer experiments: PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols were cultured in the presence or absence of the GlialCAM37o-389 peptide. The supernatant was transferred to fresh PBMCs from the same donors in the (K) absence or (L) presence of a monoclonal IL27-blocking antibody. The HLA-E MFI was assessed from PBNCs by flow cytometry. The dashed black line indicates an equal HLA-E expression before and after the transfer of the supernatant. (M) PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols were stimulated with the GlialCAM37o-389 peptide and the percentage of IL-27 expressing cells was assessed in EBV-infected (EBER+) B-cells, monocytes, NK cells, dendritic cells and non-EB V-infected (EBER ) B-cells by intracellular cytokine staining. (A-M). The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. Each dot represents one individual patient. EBER: Epstein-Barr virus (EBV)-encoded small RNAs, MFI: mean fluorescence intensity, MS: Multiple Sclerosis, ns: not significant.

[0494] Figure 13: GlialCAM37o-389-specific immune cells from MS patients upregulate HLA-E via the EBV-mediated secretion of IL-27.

[0495] (A-F) HLA-E upregulation: PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhighcontrols were stimulated with the GlialCAM37o-389-peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). The (A,C,E) percentage of HLA-E expressing cells or (B,D,F) HLA-E MFI of GlialCAM37o-389-specific (A-B) CD4+T-cells, (C-D) CD8+T-cells, and (E-F) CD138+plasma cells (GlialCAM37o-389) or total (A-B) CD4+T-cells, (C-D) CD8+T-cells, and (E-F) CD138+plasma cells (NC and NPC) was then assessed by flow-cytometry. (A-F) The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. Each dot represents one individual patient. (G-L) NKG2A inhibition: PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols were stimulated with the GlialCAM37o-389-peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). Sorted GlialCAM37o-389-specific (GlialCAM37o-389) or total (NC and NPC) CD4+T- cells, CD8+T-cells, CD138+plasma cells were co-cultured with NKG2A+NK cells from 24 healthy blood donors in the presence or absence of monoclonal NKG2A-specific blocking antibodies (Monalizumab). The IFNy concentration was then assessed by IFNy ELISA in response to (G-H) CD4+T-cells, (I- J) CD8+T-cells, or (K-L) CD138+plasma cells. (G-L) The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. Each dot represents one individual patient. (M-N) EBV reactivation: PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols were stimulated with the GlialCAM37o -389-peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). (M) The EBV viral load was assessed by EBV qPCR of the supernatant. The dashed black line indicates the detection limit of the EBV qPCR. (N) The frequency of EBV-infected (EBV EBER+) B-cells was assessed by flow cytometry. Data are shown as EBER+B-cells per IxlO7total B-cells. The groups were compared by the Kruskal-Wallis and the Dunn’s multiple comparison test. (M-N) Each dot represents one individual patient. (O) IL-27: PBMCs from N=20 MS patients, N=48 EBNAlow, and N=32 EBNAhlghcontrols were stimulated with the GlialCAM37o- 389-peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC) or without any peptides (non-peptide control, NPC). The IL-27 concentration in the supernatant was assessed by IL-27 ELISA. The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. Each dot represents one individual patient. EBER: Epstein-Barr virus (EBV)-encoded small RNAs, NC: negative control, NPC: non-peptide control, ns: not significant.

[0496] Figure 14 and Figure 14.1 A-C and J: EBV-specific immune responses in MS patients.

[0497] (A) Distribution of LMP-1 peptide variants in EBV-isolates of N=270 MS patients, N=162 EBNAlow, and N=108 EBNAhlghcontrols. Fractions represent the relative frequency of the LMP-1 peptide GGDPHLPTL, GSDPHLPTL, GGDPHI.PPI., GGDPPI.PTI., GCDPHLPTL, GIDPHLPTL, GAGPHI.PTI., GGDTPLPTL, GDDPHLPTL, GGDPHVPTL, and GTDPHLPTL variants. The frequency of the LMP-1 variant was compared between the cohorts by the Chi2Test. The individual panels in the bars relate from left to right to: GTDPHLPTL, GGDPHLPTL, GGDPPLPTL, GIDPHLPTL, GCDPHLPTL, GDDPHLPTL, GAGPHLPTL, GGDPHVPTL, GGDTPLPTL, GSDPHLPTL and GGDPHLPPL. (B-C) HLA-E stabilization and NKG2A inhibition assays. GlialCAM37o-389-specific B-cells, derived from 24 healthy individuals, were individually transfected with the indicated concentrations of the positive control (VMAPRTLIL) or the LMP-1 derived GGDPHLPTL, GSDPHLPTL, GGDPHLPPL, GGDPPLPTL, GCDPHLPTL, GIDPHLPTL, GAGPHLPTL, GGDTPLPTL, GDDPHLPTL, GGDPHVPTL, and GTDPHLPTL variants. (B) The cell surface expression of HLA-E was then analyzed by flow cytometry. (C) The transfected cells were then co-cultured with sorted NKG2A+NKG2C NKG2D' NK cells from the same donor. The inhibition of the NKG2A+NKG2C NKG2D' NK cells was assessed by IFNy ELISA. (B-C) Plots represent the mean (±SEM) of 24 independent experiments. Each peptide was compared to the positive control using the Friedman-Test. (D-H) Killing assays: (D-E) PBMCs from 60 healthy control individuals were first stimulated with the GlialCAM37o-389- peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC). Sorted GlialCAM37o -389-specific B-cells or non-GlialCAM37o-389-specific B-cells or RAJI cells were infected with the B95-8 derived EBV strain or left uninfected. (D) ADCP or (E) ADCC responses were assessed using monocyte-derived Ml macrophages and plasma from 60 healthy blood donors. (D) GlialCAM37o-389-specific ADCP was assessed as AF647 MFI by flow cytometry. (E) GlialCAM37o-389-specific ADCC was assessed as CD107a+CD14+cells by flow cytometry. (F-H) EBV-specific CD8+T-cell responses. PBMCs from 60 healthy control individuals were first stimulated with the GlialCAM37o-389-peptide (GlialCAM37o-389) or a random 20 amino acid peptide pool (negative control, NC). Sorted GlialCAM37o-389-specific B-cells or non-GlialCAM37o-389- specific B-cells or RAJI cells were infected with the B95-8 derived EBV strain or left uninfected.

[0498] (F) B-cells or RAJI cells were then co-cultured with sorted total EBV-specific CD8+T-cells from 60 healthy EBV-seropositive blood donors. The cell activation level was assessed by IFNy ELISA.

[0499] (G) The HLA-E-expression of EBV-infected or non-infected B-cells or RAJI cells was evaluated by flow-cytometry. (H) B-cells or RAJI cells were then co-cultured with sorted total EBV-specific and HLA-E restricted CD8+T-cells from 60 healthy EBV-seropositive blood donors. The cell activation level was assessed by IFNy ELISA. (D-H) The data were normalized to the corresponding non-infected control and the groups were compared by the Wilcoxon matched T- test. Each dot represents one individual sample from one healthy control. The dashed black line indicates an equal activation against infected and non-infected cells. (I) EBV-specific and HLA- E restricted CD8+T-cell frequencies were assessed in N=20 MS patients, N=48 EBNAlowand N=32 EBNAhlghcontrols by flow cytometry and compared by the Kruskal-Wallis and the Dunn’s multiple comparison test. (J) Distribution of HLA-E variants in N=270 MS patients, N=162 EBNAlowand N=108 EBNAhlghcontrols. Fractions represent the relative frequency of the HLA- E*0101 / 0101, HLA-E*0101 / 0103, and HLA-E*0103 / 0103. The frequency of the HLA-E variant was compared between the cohorts by the Chi2Test. (K-L) EBV-specific, HLA-E restricted CD8+T-cells: EBV-infected GlialCAMsvo -389-specific B-cells from 60 healthy controls, expressing either the HLA-E*0101 / 0101 (N=20), HLA-E*0101 / 0103 (N=20) or HLA-E*0103 / 0103 (N=20) genotype were cultured together with EBV-specific, HLA-E-restricted EBV-specific CD8+T- cells. (J) The activation of the EBV-specific, HLA-E-restricted EBV-specific CD8+T-cells were then assessed by IFNy ELISA. The individual panels in the bars relate from left to right to: HLA- E*0101 / 0101, HLA-E*0101 / 0103 and HLA-E*0103 / 0103. (L) The proliferation of EBV-specific, HLA-E-restricted EBV-specific CD8+T-cells were then assessed by flow cytometry. (K-L) The groups were compared by the Kruskal -Wallis and the Dunn’s multiple comparison test. Each dot represents one individual patient. The dashed black line indicates an equal activation against infected and non-infected cells. ADCC: antibody dependent cellular cytotoxicity, ADCP: antibody dependent cellular phagocytosis, MFI: mean fluorescence intensity, MS: Multiple Sclerosis, ns: not significant.

[0500] Figure 15: Forest plot for (A) individual and (B) combined risk factors in MS and healthy EBNAhlghcontrols.

[0501] The risk HLA-E*0101 allele (HLA-E*0101 / 0101 and HLA-E*0101 / 0103 genotypes) was assessed against the protective HLA-E*0103 / 0103 genotype. The risk NKG2DLNKallele (NKG2DLNK / LNKand NKG2DLNK / HNKgenotypes) was assessed against the protective NKG2DHNK / HNKgenotype. The risk EBV-encoded LMP-1 variants (GGDPHLPTL and GGDPPLPTL) were assessed against the protective GSDPHLPTL, GGDPHLPPL, GGDPHLPTL, GIDPHLPTL, GAGPHLPTL, GGDTPLPTL, GDDPHLPTL, GGDPHVPTL and GTDPHLPTL variants. The risk NKG2C phenotype (HCMV-seronegative, HCMV-seropositive and KLRC2del / deHCMV-seropositive and KLRC2'fiil'fitor KLRC2vAldeiand low binding UL40 variants (Table 11) was assessed against the protective combination HCMV-seropositive and KLRC2v vAor KLRC2'fAlddand high binding UL40 variants. The frequency of the combination was compared between the cohorts by the Chi2Test. CI: Confidence Interval, OR: Odds Ratio.

[0502] Figure 16 and Figure 16.1 A-I: EBNA-138i-452-specific antibody levels can predict the future development of MS.

[0503] (A-E) EBNA-138i-452-specific IgG titers were determined in a study cohort of 704 EBV seropositive adult and pediatric MS patients as well as a matched control group of 5381 EBV seropositive control individuals. (A) The frequency of EBNA-138i-452-specific antibody-positive individuals was compared between MS patients and controls using the %2-Test. (B) The EBNA- 138i-452-specific IgG titers of EBNA-138i-452-specific antibody-positive individuals were compared between MS patients and controls using the Mann-Whitney Test. (C) The level of EBNA-I381-452- specific antibody titers was compared between MS patients and controls using the %2-Test. (D) Receiver operating characteristic curve for the definition of high-level EBNA-138i-452-specific IgG antibody levels (OD>1.7) to discriminate between MS patients and controls. (E) The frequency of EBNA-138i-452-specific antibody-negative individuals, individuals with low-level EBNA-I381-452- specific IgG antibody levels (OD<1.69), and individuals high-level EBNA-1381-452-specific IgG antibody levels (OD>1.7) was compared between MS patients and controls using the %2-Test. (F- I) EBNA-138i-452-specific antibodies were analyzed in a study cohort of 304 EBV seropositive MS patients as well as a matched control group of 304 EBV seropositive control individuals. From each study participant, multiple plasma samples were analyzed: (1) at the time point of EBV seroconversion; (2) between 8-9 months after EBV conversion; (3) between 16-18 months after EBV conversion; (4) between 25-27 months after EBV conversion; (4) between 33-36 months after EBV conversion; and (5) at disease onset in MS patients or a matched time point after EBV conversion for healthy controls. (F) EBNA-138i-452-specific antibody titers were compared between MS patients and controls using the Mann-Whitney Test. (G) The frequency of EBNA- 138i-452-specific antibody-negative individuals, individuals with low-level EBNA-138i-452-specific IgG antibody levels (OD<1.69), and individuals high-level EBNA-138i-452-specific IgG antibody levels (OD>1.7) was compared between MS patients and controls using the %2-Test. (H) Number of consecutive high-level EBNA-138i-452-specific antibody titers (OD>1.7) was compared between MS patients and controls using the %2-Test. (I) Number of consecutive high-level EBNA-I381-452- specific antibody titers (OD>1.7) was correlated to the disease onset of MS patients using the Logrank (Mantel-Cox) test. EBV: Epstein-Barr virus; MS: Multiple sclerosis; OD: Optical Density referring to the difference in absorbance between two measurements at = 450 nm and X = 620 nm.

[0504] The invention is illustrated in the following examples:

[0505] Example 1:

[0506] Epstein-Barr Virus EBNA386-405 Peptide-specific Antibodies: A novel Predictor for the

[0507] Development of Multiple Sclerosis

[0508] List of abbreviations:

[0509] Introduction

[0510] Multiple sclerosis (MS) is a chronic autoinflammatory demyelinating disease of the central nervous system (CNS). A total of 2.8 million people are estimated to live with MS worldwide (35.9 per 100,000 population) h So far, there is no diagnostic test available that could predict the occurrence of MS. Epstein-Barr Virus (EBV) is a ubiquitous herpesvirus that persistently infects more than 90% of the adult human population worldwide2. EBV is increasingly considered an important factor in MS pathogenesis3. There is a substantially higher risk of developing MS after primary EBV infection4, and this is especially linked to the development of high-level EBV EBNA-1 -specific antibody titers in individual human hosts5. Mechanistically, this may be due to the molecular mimicry between a highly conserved region in the EBNA-1 region (EBNA381-452) that cross-react with distinct CNS-derived proteins and thus have a potential role in molecular mimicry and autoreactivity6.

[0511] We hypothesized that the EBNA38i-452-derived antibody titers are a reliable prognostic and / or diagnostic marker for the future development of MS.

[0512] Study Cohort

[0513] For this single-center case-control study, a total of N=270 EBNA-1 -seropositive MS patients and N=270 EBNA-1 -seropositive healthy controls were included. MS was diagnosed in all patients according to the McDonald criteria7All MS patients were part of the Vienna Multiple Sclerosis Database. For each patient with MS, one healthy control individual was included in the study. For all MS patients, retrospective follow-up serum samples in the range of 11-13 (median: 12.1) months were available.

[0514] Control individuals were healthy, voluntary blood donors without a history of MS or other autoimmune diseases or neuro-inflammatory disorders. Controls were matched according to sex and age to the MS patients. From all controls, retrospective follow-up serum samples, matched to the MS cohort (± 1 month) were available.

[0515] Material and Methods

[0516] Peptide-specific IgG antibodies were quantified by enzyme-linked immunosorbent assay (ELISA) as described before3. Briefly, 96-well plates (Thermo Fisher Scientific) were coated with 1 pg / mL of indicated peptides (Table 1). The coated peptides were then incubated together with 1 : 100 diluted patient or control plasma. Peptide-specific IgG antibodies were detected using HRP- conjugated goat anti-human IgG (1 :12500) and TMB substrate (both Thermo Fisher). The cut-off for detection was defined as the 95.0% CI, using 10 healthy EBV-seronegative blood donors, as described before in detail8.

[0517] Table 1: EBNA381 -452-derived peptide library Results

[0518] Individual Peptide-Specific Antibody Titers

[0519] We first assessed individual EBNA38i-452-derived peptide-specific IgG antibody titers in our study cohort of N=270 MS patients at the time point of MS diagnosis as well as of N=270 matched, healthy controls. As shown in Fig.1 and Table 2, the difference in means between MS patients and healthy controls substantially varied between the individual EBNA38i-452-derived peptides.

[0520] 18 Peptides (Peptide_20, Peptide_21, Peptide_22, Peptide_23, Peptide_24, Peptide_25, Peptide_26, Peptide_32, Peptide_33, Peptide_34, Peptide_35, Peptide_36, Peptide_37, Peptide_38, Peptide_39, Peptide_40, Peptide_52, Peptide_53) showed no signal above the detection limit of our assay. This is in contrast to WO 01 / 58481 A2 “DIAGNOSTICS AND THERAPY OF EPSTEIN-BARR VIRUS IN AUTOIMMUNE DISORDERS”, where the authors claimed an antibody response against the GPSTGPRG epitope (SEQ ID NO: 105 of WO 01 / 58481 A2). No such antibody responses were, however, detectable in our study cohort.

[0521] Peptide_07-, Peptide_08-, Peptide_09-, Peptide lO-, Peptide l l-, Peptide_12-, and Peptide_13- specific IgG antibody responses showed a high-level difference between MS patients and controls (p<0.0001).

[0522] Peptide Ol-, Peptide_02-, Peptide_03-, Peptide_04-, Peptide_05-, Peptide_06-, Peptide_14-,

[0523] Peptide_15-, Peptide_16-, Peptide_17-, Peptide_18-, Peptide_19-, Peptide_27-, Peptide_28-,

[0524] Peptide_29-, Peptide_30-, Peptide_31-, Peptide_41-, Peptide_42-, Peptide_43-, Peptide_44-,

[0525] Peptide_45-, Peptide_46-, Peptide_47-, Peptide_48-, Peptide_49-, Peptide_50-, and Peptide_51- specific IgG antibody responses showed a moderate (p<0.001) difference between MS patients and controls.

[0526] Accordingly, detecting EBNA38i-452-derived peptide-specific IgG antibodies with the individual peptides Peptide Ol, Peptide_02, Peptide_03, Peptide_04, Peptide_05, Peptide_06, Peptide_07, Peptide_08, Peptide_09, Peptide lO, Peptide l l, Peptide_12, Peptide_13, Peptide_14,

[0527] Peptide_15, Peptide_16, Peptide_17, Peptide_18, Peptide_19, Peptide_27, Peptide_28,

[0528] Peptide_29, Peptide_30, Peptide_31, Peptide_41, Peptide_42, Peptide_43, Peptide_44,

[0529] Peptide_45, Peptide_46, Peptide_47, Peptide_48, Peptide_49, Peptide_50, and Peptide_51 significantly predicts the risk of a subject of contracting MS. Table 2: Individual EBNA38i-452-derived peptide-specific antibody titers (comparison between MS patients and healthy controls)

[0530] In Table 2.1 we ranked the individual EBNA38i-452-derived peptide-specific IgG antibody titers, according to their difference of means between MS patients and controls. A high difference of means indicates a high difference in the peptide-specific antibody response between MS patients and controls. The individual peptides with a high difference of means are particularly useful for detection of EBNA38i-452-derived peptide-specific IgG antibodies thereby predicting a subject’s risk of contracting MS with unmatched confidence. We excluded all peptides that showed no signal above the detection limit of our assay. Table 2.1: Individual EBNA381 -452-derived peptide-specific antibody titers (comparison between MS patients and healthy controls)

[0531] Peptide Pools-specific antibody titers (comparison between MS patients and healthy controls)

[0532] We then aimed to define a peptide pool that is associated with the highest difference of means of peptide pool-specific IgG antibodies between MS patients and controls at the time point of MS diagnosis. Therefore, we first pooled all EBNA38i-452-derived peptides (Peptide Ol - Peptide_53) and assessed then the EBNA381-452 peptide pool-specific IgG antibody titers in our study cohort of N=270 MS patients at the time point of MS diagnosis as well as of N=270 matched, healthy controls. As shown in Table 3, the difference of means was substantial higher for the EBNA381-452 peptide pool, compared to each individual EBNA38i-452-derived peptide-specific IgG antibody titer (Table 2). These data demonstrate that the combination of the peptides (as assessed with the EBNA381-452 peptide pool of Peptides 1 to 53), can predict a subject’s risk the development of MS with very high confidence. Accordingly a peptide pool can be used in addition or instead of individual peptides for the prognosis of MS as described above.

[0533] Table 3: EBNA381 -452-derived Peptide Pool (Peptide Ol - Peptide_53) -specific antibody titers (comparison between MS patients and healthy controls)

[0534] We then aimed to assess, how the confidence to predict the occurrence of MS in a subject behaves in relation to the number of peptides in the peptide pool (Peptide Ol - Peptide_53). We performed these experiments to evaluate, how many peptides may be used to discriminate between MS patients and controls with high confidence.

[0535] Therefore, we first established one peptide pool with all peptides above the detection limit (Peptide Ol-, Peptide_02-, Peptide_03-, Peptide_04-, Peptide_05-, Peptide_06-, Peptide_07-, Peptide_08-, Peptide_09-, Peptide lO-, Peptide l l-, Peptide_12-, and Peptide_13-, Peptide_14-, Peptide_15-, Peptide_16-, Peptide_17-, Peptide_18-, Peptide_19-, Peptide_27-, Peptide_28-, Peptide_29-, Peptide_30-, Peptide_31-, Peptide_41-, Peptide_42-, Peptide_43-, Peptide_44-, Peptide_45-, Peptide_46-, Peptide_47-, Peptide_48-, Peptide_49-, Peptide_50-, and Peptide_51) and all peptides below the detection limit (Peptide_20, Peptide_21, Peptide_22, Peptide_23, Peptide_24, Peptide_25, Peptide_26, Peptide_32, Peptide_33, Peptide_34, Peptide_35, Peptide_36, Peptide_37, Peptide_38, Peptide_39, Peptide_40, Peptide_52, Peptide_53) and assessed EBNA381-452 peptide pool-specific IgG antibody titers for both pools using our study cohort of N=270 MS patients at the time point of MS diagnosis and of N=270 matched, healthy controls. As shown in Table 4, the peptide pool with all peptides above the detection limit (N=35 peptides) showed comparable difference of means than the entire peptide pool, consisting of all (N=53) EBNA38i-452-derived peptides (see: Table 3). Accordingly, both pools are capable of predicting the occurrence of MS in a subject with high confidence. Table 4: EBNA38i-452-derived Peptide Subpool-specific antibody titers (comparison between MS patients and healthy controls)

[0536] We then aimed to assess how reducing the number of peptides in the peptide pool affects the confidence to predict the occurrence of MS in a subject. For this, we gradually reduced the number of peptides in the peptide pool. Therefore, we selected all peptides above the detection limit (Peptide Ol, Peptide_02, Peptide_03, Peptide_04, Peptide_05, Peptide_06, Peptide_07, Peptide_08, Peptide_09, Peptide lO, Peptide l l, Peptide_12, Peptide_13, Peptide_14,

[0537] Peptide_15, Peptide_16, Peptide_17, Peptide_18, Peptide_19, Peptide_27, Peptide_28,

[0538] Peptide_29, Peptide_30, Peptide_31, Peptide_41, Peptide_42, Peptide_43, Peptide_44,

[0539] Peptide_45, Peptide_46, Peptide_47, Peptide_48, Peptide_49, Peptide_50, and Peptide_51) and established individual peptide pools with a decreasing amount of peptides. For each peptide pool, we removed one or two peptides with the lowest differences of mean (see Table 2), respectively. We then assessed EBNA381-452 peptide pool-specific IgG antibody titers for all pools using our study cohort of N=270 MS patients at the time point of MS diagnosis and of N=270 matched, healthy controls. As shown in Table 5, a peptide pool with already >2 peptides was associated with a highly significant prediction of MS. Therefore, a peptide pool comprising >2 peptides capable of predicting MS in a subject with high confidence.

[0540] Table 5: EBNA381 -452-derived Peptide Pool-specific antibody titers (comparison between MS patients and healthy controls)

[0541] As shown in Table 5, a peptide pool with already >2 peptides (Peptide_09 and Peptide_10)-specific IgG antibody titers was associated with a highly significant prediction of MS. These results show that individual peptides as well as peptide pools of >2 peptides can highly significantly predict MS in a subject resulting in a high confidence prognosis.

[0542] Comparison of our peptide pools to Peptide 09 From all peptides, reported in WO 98 / 030586 A2 and WO 01 / 58481 A2, Peptide_9-specific IgG antibodies had the highest predictive value for the diagnosis of MS. Accordingly, Peptide_9 may be used as a benchmark to compare the significance of the peptide pools to predict MS in a subject with the prior art. We therefore compared the predictive capacity of all our peptide pools against Peptide_9-specific IgG antibodies. As shown in Table 6, already the presence of two peptide- specific antibodies (Peptide_09 and Peptide lO) was associated with a higher significance resulting in a higher confidence for predicting MS in a subject than e.g. using Peptide_9-specific antibodies alone (p<0.04).

[0543] Table 6: Comparison to WO 1998030586 and WO0158481A2 A value of <0.04 corresponds to all p-values between p<0.0001 and p<0.04. Ill

[0544] We then tested, whether a specific combination of two peptide-specific IgG antibody levels is associated with an improved prediction for MS in a subject than Peptide_9specific IgG antibodies alone. Therefore, we first selected Peptide_03, Peptide_14, Peptide_29, and Peptide_48 that had no overlapping antigenic epitopes and represent the highest difference in means between MS patients and healthy controls from each individual peptide-specific antibody peak (Fig.1). We then compared these selected EBNA38i-452-derived peptide pool-specific antibody responses against Peptide_09 specific antibody titers. As shown in Table 7, the presence of two EBNA38i-452-derived peptide pool-specific antibody responses was associated with a better predication than Peptide_9 alone.

[0545] Table 7: Comparison to WO1998030586 and WO0158481A2

[0546] A value of <0.04 corresponds to all p-values between p<0.0001 and p<0.04.

[0547] Prediction of MS

[0548] We then assessed whether EBNA381-452 peptide pool-specific IgG antibody titers could serve as a prognostic marker for the development of MS. Therefore, we collected plasma samples from MS patients between 1-2 years as well as between 3-4 years prior to MS diagnosis. From healthy controls, we collected plasma samples at a matched time point. We then tested all plasma samples with EBNA38i-452-derived Peptide pool, containing all 53 EBNA38i-452-derived peptides. As shown in Fig. 2, MS patients had, compared to healthy controls, significantly higher EBNA381-452 peptide pool-specific IgG antibody titers, at 1-2 years or 3-4 years prior to MS diagnosis. Accordingly, the peptides provided herein can be used to predict if a subject is at risk of contracting MS 1-2 years or 3-4 years prior to MS diagnosis. This facilitates an early diagnosis of MS, e.g. via frequent medical examination to detect MS symptoms early. This in turn allows for early onset of treatment which can prevent the progression of MS and further disability of the patient.

[0549] List of references general description and Example 1:

[0550] 1. Walton, C., et al. Rising prevalence of multiple sclerosis worldwide: Insights from the Atlas of MS, third edition. Mult Scler 26, 1816-1821 (2020).

[0551] 2. Dunmire, S.K., Verghese, P.S. & Balfour, H.H., Jr. Primary Epstein-Barr virus infection. Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology 102, 84-92 (2018).

[0552] 3. Vietzen, H., et al. Ineffective control of Epstein-Barr- virus-induced autoimmunity increases the risk for multiple sclerosis. Cell 186, 5705-5718. e5713 (2023).

[0553] 4. Bjornevik, K., et al. Longitudinal analysis reveals high prevalence of Epstein-Barr virus associated with multiple sclerosis. Science (New York, N.Y.) 375, 296-301 (2022).

[0554] 5. Levin, L.I., et al. Temporal Relationship Between Elevation of Epstein-Barr Virus Antibody Titers and Initial Onset of Neurological Symptoms in Multiple Sclerosis. JAMA 293, 2496-2500 (2005).

[0555] 6. Soldan, S.S. & Lieberman, P.M. Epstein-Barr virus and multiple sclerosis. Nature reviews. Microbiology 21, 51-64 (2023).

[0556] 7. Thompson, A. J., et al. Diagnosis of multiple sclerosis: 2017 revisions of the McDonald criteria. Lancet Neurol 17, 162-173 (2018).

[0557] 8. Frey, A., Di Canzio, J. & Zurakowski, D. A statistically defined endpoint titer determination method for immunoassays. Journal of immunological methods 221, 35-41 (1998).

[0558] 9. Waubant E. Improving outcomes in multiple sclerosis through early diagnosis and effective management. Prim Care Companion CNS Disord. 2012;14(5):PCC.11016co2cc. 10. Tobin WO. Early Diagnosis and Treatment Are Associated With Improved Outcomes in Patients With Multiple Sclerosis. Neurology. 2021 Oct 26;97(17):799-800.

[0559] 11. Wiendl H, Gold R, Berger T, Derfuss T, Linker R, Maurer M, et al. [Multiple sclerosis treatment consensus group (MSTCG): position paper on disease-modifying treatment of multiple sclerosis 2021 (white paper)]. Nervenarzt. 2021 Aug;92(8):773-801.

[0560] 12. Goodin DS, Bates D. Treatment of early multiple sclerosis: the value of treatment initiation after a first clinical episode. Mult Scler. 2009 Oct;15(10): 1175-82.

[0561] 13. Goodin DS, Reder AT, Bermel RA, Cutter GR, Fox RJ, John GR, et al. Relapses in multiple sclerosis: Relationship to disability. Mult Scler Relat Disord. 2016 Mar;6: 10-20.

[0562] 14. Kavaliunas A, Manouchehrinia A, Stawiarz L, Ramanujam R, Agholme J, Hedstrbm AK, et al. Importance of early treatment initiation in the clinical course of multiple sclerosis. Mult Scler. 2017 Aug;23(9): 1233-40.

[0563] 15. Spelman T, Magyari M, Piehl F, Svenningsson A, Rasmussen PV, KantM, et al. Treatment Escalation vs Immediate Initiation of Highly Effective Treatment for Patients With Relapsing-Remitting Multiple Sclerosis: Data From 2 Different National Strategies. JAMA Neurol. 2021 Oct l;78(10): 1197-204.

[0564] 16. Guger M, Enzinger C, Leutmezer F, Di Pauli F, Kraus J, Kalcher S, et al. Early intensive versus escalation treatment in patients with relapsing-remitting multiple sclerosis in Austria. J Neurol. 2024 Mar 2.

[0565] 17. Uher T, Adzima A, Srpova B, Noskova L, Marechai B, Maceski AM, et al. Diagnostic delay of multiple sclerosis: prevalence, determinants and consequences. Mult Scler. 2023 Oct;29(l 1-12): 1437-51.

[0566] 18. Stankiewicz JM, Weiner HL. An argument for broad use of high efficacy treatments in early multiple sclerosis. Neurol Neuroimmunol Neuroinflamm. 2020 Jan;7(l):e636.

[0567] 19. Karampampa K, Gyllensten H, Murley C, Alexanderson K, Kavaliunas A, Olsson T, et al. Early vs. late treatment initiation in multiple sclerosis and its impact on cost of illness: A register-based prospective cohort study in Sweden. Mult Scler J Exp Transl Clin. 2022;8(2):20552173221092411....

Claims

CLAIMS1. A peptide comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or a variant of said peptide having at least about 60% sequence identity to the peptide.

2. The peptide of claim 1 comprising 16 to 72, preferably 16 to 20, consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69.

3. The peptide of claim 1 comprising 5 to 38 consecutive amino acids of the sequence set forth in SEQ ID NO: 55, or comprising 5 to 24 consecutive amino acids of the sequence set forth in SEQ ID NO: 56, or comprising 5 to 30 consecutive amino acids of the sequence set forth in SEQ ID NO: 57, or a variant of said peptide having at least about 60% sequence identity to the peptide.

4. The peptide of any one of claims 1 to 3, wherein the consecutive amino acids are, comprise and / or form a functional epitope.

5. The peptide of any one of claims 1 to 4, wherein the peptide is capable of eliciting an autoimmune response, and / or wherein the peptide is capable of binding antibodies or being bound by antibodies that cross-react with central nervous system (CNS)-derived proteins.

6. The peptide of any one of claims 1 to 5, wherein the peptide is selected from the group consisting of SEQ ID NOs: 1-53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

7. The peptide of any one of claims 1 to 6, wherein the peptide is selected from the group consisting of SEQ ID NOs: 1-19, 27-31 and 41-51, or a variant of said peptide having at least about 60% sequence identity to the peptide.

8. The peptide of any one of claims 1 to 7, wherein the peptide is the peptide of SEQ ID NO: 9, or a variant of said peptide having at least about 60% sequence identity to the peptide.

9. The peptide of any one of claims 1 to 8, wherein the peptide is comprised in a solid phase, a semi-solid phase or liquid phase.

10. The peptide of claim 9, wherein the peptide is immobilized in or on a semi-solid or solid surface.

11. The peptide of claim 10, wherein the peptide is coated onto the surface of a membrane, microtiter plate, bead, gel, or column or is immobilized in a gel.

12. Two or more peptides of any one of claims 1 to 11, wherein the peptides are covalently or partially covalently conjugated to each other.

13. A composition comprising at least two peptides each comprising 5 to 72 consecutive amino acids of the sequence set forth in SEQ ID NO: 54, and / or in any one of SEQ ID NO: 63-69, or variants of said peptides having at least about 60% sequence identity to the peptides.

14. The composition of claim 13 comprising at least two peptides each comprising 5 to 38 consecutive amino acids of the sequence set forth in SEQ ID NO: 55, and / or comprising 5 to 24 consecutive amino acids of the sequence set forth in SEQ ID NO: 56, and / or comprising 5 to 30 consecutive amino acids of the sequence set forth in SEQ ID NO: 57, or variants of said peptides having at least about 60% sequence identity to the peptides.

15. The composition of claim 13 or 14, wherein the consecutive amino acids are, comprise and / or form a functional epitope.

16. The composition of any one of claims 13 to 15, wherein the at least two peptides are capable of eliciting an autoimmune response, and / or wherein the at least two peptides are capable of binding antibodies or being bound by antibodies that cross-react with central nervous system (CNS)-derived proteins.

17. The composition of any one of claims 13 to 16 comprising at least two peptides selected from the group of SEQ ID NOs: 1-53, or variants of said at least two peptides having at least about 60% sequence identity to the peptides.

18. The composition of any one of claims 13 to 17, comprising at least two peptides selected from the group of SEQ ID NOs: 1-19, 27-31 and 41-51, or variants of said at least two peptides having at least about 60% sequence identity to the peptides.

19. The composition of any one of claims 13 to 18, comprising(i) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or(ii) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or(iii) at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, or(iv) at least one peptide selected from the group of SEQ ID NOs: 1-19 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 27-31 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide, and at least one peptide selected from the group of SEQ ID NOs: 41-51 or a variant of said at least one peptide having at least about 60% sequence identity to the peptide.

20. The composition of any one of claims 13 to 19, comprising the peptides of SEQ ID NOs: 1- 19 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 27-31 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 or variants of said peptideshaving at least about 60% sequence identity to the peptides.

21. The composition of claim 20, consisting of the peptides of SEQ ID NOs: 1-19 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 27-31 or variants of said peptides having at least about 60% sequence identity to the peptides, and / or the peptides of SEQ ID NOs: 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier, preferably consisting of the peptides of SEQ ID NOs: 1-19, 27-31 and 41-51 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier.

22. The composition of any one of claims 13 to 20, comprising the peptides of SEQ ID NOs: 1- 53 or variants of said peptides having at least about 60% sequence identity to the peptides.

23. The composition of claim 22 consisting of the peptides of SEQ ID NOs: 1-53 or variants of said peptides having at least about 60% sequence identity to the peptides and a suitable carrier.

24. The composition of any one of claims 13 to 18 comprising the peptides of:(a) SEQ ID NOs: 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or(b) SEQ ID NOs: 8, 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or(c) SEQ ID NOs: 8-12 or variants of said peptides having at least about 60% sequence identity to the peptides, or(d) SEQ ID NOs: 7-13 or variants of said peptides having at least about 60% sequence identity to the peptides, or(e) SEQ ID NOs: 7-13, 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(f) SEQ ID NOs: 7-14, 29, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(g) SEQ ID NOs: 7-15, 29, 43, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(h) SEQ ID NOs: 3, 7-15, 29, 43, 45, 47 and 48 or variants of said peptides having at leastabout 60% sequence identity to the peptides, or(i) SEQ ID NOs: 3-4, 7-15, 29, 43, and 45-48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(j) SEQ ID NOs: 3-16, 29, 43, and 45-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or(k) SEQ ID NOs: 3-17, 29, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or(l) SEQ ID NOs: 3-17, 28-30, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or(m) SEQ ID NOs: 3-17, 19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(n) SEQ ID NOs: 2-19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(o) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(p) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(q) SEQ ID NOs: 2-19, 27-31, and 42-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination of the above.

25. The composition of claim 24, consisting of the peptides of:(a) SEQ ID NOs: 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or(b) SEQ ID NOs: 8, 9 and 10 or variants of said peptides having at least about 60% sequence identity to the peptides, or(c) SEQ ID NOs: 8-12 or variants of said peptides having at least about 60% sequence identity to the peptides, or(d) SEQ ID NOs: 7-13 or variants of said peptides having at least about 60% sequence identity to the peptides, or(e) SEQ ID NOs: 7-13, 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(f) SEQ ID NOs: 7-14, 29, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(g) SEQ ID NOs: 7-15, 29, 43, 45 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(h) SEQ ID NOs: 3, 7-15, 29, 43, 45, 47 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(i) SEQ ID NOs: 3-4, 7-15, 29, 43, and 45-48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(j) SEQ ID NOs: 3-16, 29, 43, and 45-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or(k) SEQ ID NOs: 3-17, 29, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or(l) SEQ ID NOs: 3-17, 28-30, and 43-49 or variants of said peptides having at least about 60% sequence identity to the peptides, or(m) SEQ ID NOs: 3-17, 19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(n) SEQ ID NOs: 2-19, 28-30, 43-49 and 51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(o) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(p) SEQ ID NOs: 2-19, 27-30, and 43-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or(q) SEQ ID NOs: 2-19, 27-31, and 42-51 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination thereof and a suitable carrier.

26. The composition of any one of claims 13 to 18 comprising the peptides of:(i) SEQ ID NOs: 3 and 14 or variants of said peptides having at least about 60% sequence identity to the peptides, or(ii) SEQ ID NOs: 3 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or(iii) SEQ ID NOs: 3 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(iv) SEQ ID NOs: 14 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or(v) SEQ ID NOs: 14 and 48 or variants of said peptides having at least about 60% sequenceidentity to the peptides, or(vi) SEQ ID NOs: 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination of the above.

27. The composition of claim 26 consisting of the peptides of:(i) SEQ ID NOs: 3 and 14 or variants of said peptides having at least about 60% sequence identity to the peptides, or(ii) SEQ ID NOs: 3 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or(iii) SEQ ID NOs: 3 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(iv) SEQ ID NOs: 14 and 29 or variants of said peptides having at least about 60% sequence identity to the peptides, or(v) SEQ ID NOs: 14 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or(vi) SEQ ID NOs: 29 and 48 or variants of said peptides having at least about 60% sequence identity to the peptides, or any combination thereof and a suitable carrier.

28. The composition of any one of claims 13 to 27, wherein the peptides are comprised in a solid phase, a semi-solid phase or liquid phase.

29. The composition of claim 28, wherein the peptides are immobilized in or on a semi-solid or solid surface.

30. The composition of claim 29, wherein the peptides are coated onto the surface of a membrane, microtiter plate, bead, gel or column or are immobilized in a gel.

31. The composition of any one of claims 13 to 30, wherein the at least two peptides are covalently or partially covalently conjugated to each other.

32. An antigen comprising or consisting of the peptide of any one of claims 1 to 12, or comprising or consisting of the composition of any one of claims 13 to 31.

33. A method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably multiple sclerosis (MS), the method comprising detecting and / or quantifying binding of antibodies to the peptide of any one of claims 1 to 12, or the composition of any one of claims 13 to 31, or the antigen of claim 32 in a biological sample of said subject.

34. A method for early prognosis of a subj ect being at risk of contracting an autoimmune disease, preferably multiple sclerosis (MS), the method comprising:(a) contacting a biological sample of said subject with the peptide of any one of claims 1 to 12, or the composition of any one of claims 13 to 31, or the antigen of claim 32,(b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

35. An in vitro or ex vivo method for early prognosis of a subject being at risk of contracting an autoimmune disease, preferably multiple sclerosis (MS), the method comprising:(a) contacting a biological sample of said subject with the peptide of any one of claims 1 to 12, or the composition of any one of claims 13 to 31, or the antigen of claim 32,(b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

36. The method of any one of claims 33 to 35, wherein the method is performed between about 10 years and about 1 month prior to a diagnosis or manifestation of the autoimmune disease in the subject, preferably, wherein the method is performed at least 4, 3, 2, or at least 1 year prior to a diagnosis or manifestation of the autoimmune disease in the subject.

37. The method of any one of claims 33 to 36, wherein the method is performed between about 10 years and about 1 month after Epstein-Barr virus (EBV) seroconversion in the subject, preferably, wherein the method is performed between about 1 month and about 48 months after Epstein-Barr virus (EBV) seroconversion in the subject.

38. The method of any one of claims 33 to 37, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about every 1-12 months, such as every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 month(s).

39. The method of any one of claims 33 to 38, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or about 4 times after EBV seroconversion in the subject.

40. The method of any one of claims 33 to 39, wherein the autoimmune disease is selected from multiple sclerosis (MS), myasthenia gravis (MG), systemic lupus erythematosus (SLE), or autoimmune encephalitis.

41. The method of any one of claims 33 to 40, wherein the subject is at risk of contracting the autoimmune disease if binding of antibodies to said peptide, composition or antigen is detected and / or quantified.

42. The method of any one of claims 33 to 41, wherein the method further comprises step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control.

43. The method of claim 42, wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more healthy subject (negative control), and / or wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more subjects that have contracted the autoimmune disease (positive control).

44. The method of any one of claims 33 to 43, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject significantly deviates from the negative control, and / or wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject does not significantly deviate from the positive control.

45. The method of claim 44, wherein the significant deviation from the negative control is an increase in the detected and / or quantified binding in said biological sample of said subject compared to the negative control.

46. The method of any one of claims 33 to 45, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of = 450 nm and = 620 nm.

47. The method of any one of claims 33 to 45, wherein the subject is not at risk or at low risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

48. The method of any one of claims 33 to 45, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 8 years after EBV seroconversion if(i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or at least about 4 times after EBV seroconversion in the subject, and(ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

49. The method of any one of claims 33 to 45, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion if(i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 2 or about 3 times after EBV seroconversion in the subject, and(ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

50. The method of any one of claims 33 to 45, wherein the subject is not at risk or at low risk of contracting the autoimmune disease in about 8 to about 13 years after EBV seroconversion if(i) no binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified after EBV seroconversion in the subject, and / or(ii) the detected and / or quantified binding in said biological sample of said subjectcorresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of = 450 nm and = 620 nm.

51. The method of any one of claims 33 to 50, wherein the method further comprises advising pre-clinical monitoring to avoid symptomatic manifestation of an autoimmune disease, and / or to allow early diagnosis of the autoimmune disease, e.g. MS, and / or wherein the method further comprises advising early treatment, if the subject is at risk of contracting the autoimmune disease and / or wherein the method further comprises advising a delay of treatment of the autoimmune disease, if the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

52. The method of claim 51, wherein the pre-clinical monitoring comprises frequent medical examination, such as neurological and radiological monitoring of the subject, e.g. via magnetic resonance imaging (MRI) of the subject’s brain and spinal cord, lumbar puncture, optical coherence tomography (OCT) and / or electrophysiological examination.

53. The method of any one of claims 33, 34 or 36 to 52, wherein the method further comprises treatment of the autoimmune disease if the subject is at risk of contracting the autoimmune disease or, wherein the method further comprises delaying of treatment of the autoimmune disease, if the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

54. The method of any one of claims 33 to 53, wherein the binding of antibodies is detected and / or quantified using Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC), Flow Cytometry and / or Fluorescence-Activated Cell Sorting (FACS) and the like.

55. The method of any one of claims 33 to 54, wherein the biological sample is a blood sample or a cerebrospinal fluid (CSF) sample.

56. The method of claim 55, wherein the blood sample is a whole blood sample, a blood serum sample, or a blood plasma sample.

57. The method of any one of claims 33 to 56, wherein the biological sample or blood sample or cerebrospinal fluid (CSF) sample is a human biological sample or a human blood sample or a human cerebrospinal fluid (CSF) sample.

58. The peptide of any one of claims 1 to 12, or the composition of any one of claims 13 to 31, or the antigen of claim 32, for use in a method of diagnosis of a risk of contracting an autoimmune disease, preferably multiple sclerosis (MS).

59. The peptide, composition or antigen for use of claim 58, wherein the method comprises:(a) contacting a biological sample of a subject with the peptide any one of claims 1 to 12, or the composition of any one of claims 13 to 31, or the antigen of claim 32,(b) detecting and / or quantifying binding of antibodies to said peptide, composition or antigen.

60. The peptide, composition or antigen for use of claim 58 or 59, wherein the method is performed between about 10 years and about 1 month prior to a diagnosis or manifestation of the autoimmune disease in the subject, preferably, wherein the method is performed at least 4, 3, 2, or at least 1 year prior to the manifestation of the autoimmune disease in the subject.

61. The peptide, composition or antigen for use of any one of claims 58 to 60, wherein the method is performed between about 10 years and about 1 month after Epstein-Barr virus (EBV) seroconversion in the subject, preferably, wherein the method is performed between about 1 month and about 48 months after Epstein-Barr virus (EBV) seroconversion in the subject.

62. The peptide, composition or antigen for use of any one of claims 58 to 61, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about every 1-12 months, such as every 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 month(s).

63. The peptide, composition or antigen for use of any one of claims 58 to 62, wherein the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or about 4times after EBV seroconversion in the subject.

64. The peptide, composition or antigen for use of any one of claims 58 to 63, wherein the autoimmune disease is selected from multiple sclerosis (MS), myasthenia gravis (MG), or systemic lupus erythematosus (SLE) or autoimmune encephalitis.

65. The peptide, composition or antigen for use of any one of claims 58 to 64, wherein the subject is at risk of contracting the autoimmune disease if binding of antibodies to said peptide, composition or antigen is detected and / or quantified.

66. The peptide, composition or antigen for use of any one of claims 58 to 65, wherein the method further comprises step (c) comparing the detected and / or quantified binding to a control such as a positive or negative control.

67. The peptide, composition or antigen for use of claim 66, wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more healthy subject (negative control), and / or wherein the control is the detected and / or quantified binding of antibodies to said peptide, composition or antigen in a biological sample of one or more subjects that have contracted the autoimmune disease (positive control).

68. The peptide, composition or antigen for use of any one of claims 58 to 67, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject significantly deviates from the negative control, and / or wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject does not significantly deviate from the positive control.

69. The peptide, composition or antigen for use of claim 68, wherein the significant deviation from the negative control is an increase in the detected and / or quantified binding in said biological sample of said subject compared to the negative control.

70. The peptide, composition or antigen for use of any one of claims 58 to 69, wherein the subject is at risk of contracting the autoimmune disease if the detected and / or quantifiedbinding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of = 450 nm and = 620 nm.

71. The peptide, composition or antigen for use of any one of claims 58 to 69, wherein the subject is not at risk or at low risk of contracting the autoimmune disease if the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA- 138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

72. The peptide, composition or antigen for use of any one of claims 58 to 69, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 8 years after EBV seroconversion if(i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 1 to about 20, preferably about 1, 2, 3 or at least about 4 times after EBV seroconversion in the subject, and(ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

73. The peptide, composition or antigen for use of any one of claims 58 to 69, wherein the subject is at risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion if(i) the binding of antibodies to said peptide, composition or antigen in a biological sample of said subject is detected and / or quantified about 2 or about 3 times after EBV seroconversion in the subject, and(ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD>1.7, optionally, measured at a wavelength of X = 450 nm and X = 620 nm.

74. The peptide, composition or antigen for use of any one of claims 58 to 69, wherein the subject is not at risk or at low risk of contracting the autoimmune disease in about 8 to about 13 years after EBV seroconversion if(i) no binding of antibodies to said peptide, composition or antigen in a biological sample ofsaid subject is detected and / or quantified after EBV seroconversion in the subject, and / or (ii) the detected and / or quantified binding in said biological sample of said subject corresponds to an EBNA-138i-452-specific IgG antibody titer of OD<1.69, optionally, measured at a wavelength of = 450 nm and = 620 nm.

75. The peptide, composition or antigen for use of any one of claims 58 to 74, wherein the method further comprises advising pre-clinical monitoring to avoid symptomatic manifestation of an autoimmune disease, and / or to allow early diagnosis of the autoimmune disease, e.g. MS, and / or wherein the method further comprises advising early treatment, if the subject is at risk of contracting the autoimmune disease, and / or wherein the method further comprises advising a delay of treatment of the autoimmune disease, if the subject is diagnosed with a risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

76. The peptide, composition or antigen for use of claim 75, wherein the pre-clinical monitoring comprises frequent medical examination, such as neurological and radiological monitoring of the subject, e.g. via magnetic resonance imaging (MRI) of the subject’s spinal cord and / or brain, lumbar puncture, optical coherence tomography (OCT) and / or electrophysiological examination.

77. The peptide, composition or antigen for use of any one of claims 58 to 76, wherein the method further comprises treatment of the autoimmune disease if the subject is diagnosed with a risk of contracting the autoimmune disease or, wherein the method further comprises delaying treatment of the autoimmune disease, if the subject is diagnosed with a risk of contracting the autoimmune disease in about 3 to about 13 years after EBV seroconversion.

78. The peptide, composition or antigen for use of any one of claims 58 to 77, wherein the binding of antibodies is detected and / or quantified using any one of Enzyme-Linked Immunosorbent Assay (ELISA), sandwich ELISA, Enzyme-Linked Immunospot (ELISPOT), Western Blot (WB), Immunoprecipitation (IP), Immunocytochemistry (ICC) or Flow Cytometry and Fluorescence-Activated Cell Sorting (FACS) and the like.

79. The peptide, composition or antigen for use of any one of claims 59 to 78, wherein the biological sample is a blood sample or a cerebrospinal fluid (CSF) sample.

80. The peptide, composition or antigen for use of claim 79, wherein the blood sample is a whole blood sample, a blood serum sample or a blood plasma sample.

81. The peptide, composition or antigen for use of any one of claims 59 to 80, wherein the biological sample or blood sample or cerebrospinal fluid (CSF) sample is a human biological sample or a human blood sample or a human cerebrospinal fluid (CSF) sample.

82. A peptide selected from the group consisting of SEQ ID NOs: 20-26, 32-40 and 52-53, or a variant of said peptide having at least about 60% sequence identity to the peptide.

83. A composition comprising or consisting of the peptides of SEQ ID NOs: 20-26 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 32-40 or variants of said peptides having at least about 60% sequence identity to the peptides and / or the peptides of SEQ ID NOs: 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides, preferably comprising or consisting of the peptides of SEQ ID NOs: 20-26, 32-40 and 52-53 or variants of said peptides having at least about 60% sequence identity to the peptides.

84. Use of the peptide of any one of claims 1 to 12, or of the composition of any one of claims 13 to 31, or of the peptide of claim 82, or of the composition of claim 83, as an antigen.

85. A kit comprising the peptide of any one of claims 1 to 12, or the composition of any one of claims 13 to 31, or the antigen of claim 32, and / or the peptide of claim 82, and / or the composition of claim 83, and means for detecting and / or quantifying antibodies binding to said peptide, composition, or antigen.

86. The kit of claim 85, further comprising an anti-human Immunoglobulin G (IgG) antibody.

87. The kit of claim 85 or 86, wherein the means for detecting and / or quantifying antibodies binding to said peptide, composition or antigen are means to perform an Enzyme-Linked Immunosorbent Assay (ELISA).