ES2648694A1 - Proinflammatory cytokines as a diagnostic marker in the episodic vestibular syndrome. (Machine-translation by Google Translate, not legally binding) - Google Patents

Abstract

Proinflammatory cytokines as a diagnostic marker in the episodic vestibular syndrome. Use of proinflammatory cytokines il1 β, il6, tnf α to obtain useful data for the diagnosis and classification of a disease that presents with episodic vestibular syndrome, method of obtaining useful data for the diagnosis and classification of said disease, kit or device and uses. (Machine-translation by Google Translate, not legally binding)

Description

Proinflammatory cytokines as a diagnostic marker in episodic vestibular syndrome.

The present invention is within the field of biomedicine and biotechnology, and refers to the use of proinflammatory cytokines as a diagnostic marker in episodic vestibular syndrome, and especially refers to a method for the classification of patients presenting with episodic vestibular syndrome with migraine and / or hearing loss, and specifically the differential diagnosis of vestibular migraine and Meniere's disease.

STATE OF THE PREVIOUS TECHNIQUE

The classification of episodic vestibular syndrome (EVS) is based on clinical symptoms and there are no biological markers for diagnosis. Currently, consensus diagnoses have been developed for the most frequent causes of spontaneous vertigo that are: vestibular migraine (MV) and Meniere's disease (MS). However, the diagnostic criteria of these entities are not exclusive and the same patient can meet diagnostic criteria for both diseases. In addition, there are patients with an incomplete phenotype that do not meet the clinical criteria and that could be partial forms of the disease. MS is a chronic disorder that affects the inner ear, characterized by recurrent episodes of vertigo, progressive instability, hearing pressure, tinnitus and hearing loss at low and medium frequencies. There are several epidemiological evidences that indicate that MS could have a genetic basis, as well as the role of the immune response and allergy. In this sense, patients with MS have a high prevalence of autoimmune diseases as well as allergic symptoms. However, there is no biological marker to identify which patients with MS have an impaired immune response. Thus, nonspecific autoantibodies such as antinuclear antibodies or circulating immunocomplexes are only elevated in a very low percentage of patients and have no practical diagnostic profitability.

Autoimmune inner ear disease (EAOI) is a chronic disorder of the inner ear that is caused by an immune attack on the inner ear, mediated by antibodies or immunologically active cells. It is characterized by bilateral sensorineural hearing loss, rapidly progressive and often fluctuating, which occurs over a period of weeks to years. Vestibular symptoms, such as instability

Generalized, ataxia, positional vertigo and episodic vertigo, may be present in almost 50% of patients. Occasionally, only one ear is initially affected, but bilateral hearing loss occurs in most patients, with symmetric or asymmetric audiometric thresholds. Almost 25% to 50% of patients also have tinnitus and a sensation of otic fullness, which can be fluctuating.

A wide variety of treatments, both medical and surgical, have been proposed for hearing loss, and specifically for MS. All of them with different results regarding the remission of symptoms and side effects.

Among the medical treatments are the hyposodic diet, antihistamines, diuretics, subcutaneous histamine, antivertiginous drugs, benzodiazepines and transtympanic therapy, especially in that group of patients who do not respond to the usual therapies. However, these treatments have very limited efficacy, due to the clinical heterogeneity of patients with MS.

In some centers, intratympanic injection of ototoxic drugs, such as gentamicin, into the middle ear cavity has been practiced for some time. The objective of this procedure would be to produce a medical labytectomy, which would cause the decrease or disappearance of symptoms. The risk of this procedure is the deterioration of the patient's hearing capacity, especially described in those patients undergoing treatment regimens in short time periods. The most commonly used agents have been, initially, streptomycin and, currently, gentamicin.

In very severe cases, surgery has been resorted to, but this resource worsens the patient's quality of life because it eliminates not only the symptoms, but also the sense of balance.

Therefore, the treatments currently available for hearing loss can be excessively aggressive, such as ablative surgery (vestibular neurectomy or labyrinthyctomy) or the application of intratympanic gentamicin, and can put the patient's hearing at risk. The identification of an autoimmune variant through a high proinflammatory cytokine profile would allow immunosuppressive treatment in this subgroup of patients. In addition, a correct differential diagnosis between MV and MS would allow the administration of appropriate therapies, avoiding the aggressive treatments described above in misdiagnosed patients.

BRIEF DESCRIPTION OF THE INVENTION

Examples of the invention show that IL1β, IL6 and TNFα are useful markers for differentiate a disease with episodic vestibular syndrome (attacks of 5 vertigo) with migrainous symptoms (headache and / or sensory aura) and / or sensorineural hearing loss, preferably MS and MV, and even more preferably, autoimmune MS.

Therefore, a first aspect of the invention relates to the use of the proinflammatory cytokines IL1β, IL6, TNFα or any combination thereof to obtain 10 data useful for the diagnosis and classification of a disease that develops with episodic vestibular syndrome. In a preferred embodiment, the proinflammatory cytokines IL1β, IL6 and TNFα are used simultaneously. In another preferred embodiment, the determination of proinflammatory cytokines is performed in vitro in human mononuclear cells. In another even more preferred embodiment, the classification of the disease with syndrome

15 vestibular allows obtaining useful data for differential diagnosis between MS and MV, and more specifically, for the diagnosis of autoimmune MS.

MS may present with migrainous symptoms, including headache and MV may have sensorineural hearing loss, being indistinguishable in the early stages of the disease.

A second aspect of the invention relates to a method of obtaining useful data for the diagnosis and classification of a disease that develops with episodic vestibular syndrome, hereafter referred to as the first method of the invention, comprising:

A) detect the levels of IL1β, IL6 and TNFα proinflammatory cytokines in an isolated biological sample from an individual. b) compare the levels of proinflammatory cytokines detected in step (a), with baseline levels in a healthy individual.

In a preferred embodiment of this aspect the invention, the sample isolated in step (a) is peripheral blood, and even more preferably they are mononuclear cells obtained from peripheral blood.

In another even more preferred embodiment, the classification of the disease with vestibular syndrome allows obtaining useful data for the differential diagnosis between MS and MV, and more specifically, for the diagnosis of autoimmune MS.

A third aspect of the invention relates to a method for the diagnosis and classification of a disease that develops with episodic vestibular syndrome, hereafter referred to as the second method of the invention, comprising steps (a) and (b) of the first method. of the invention, and also comprises:

c) include the individual in the group of individuals with MS of autoimmune origin.

In a preferred embodiment of this aspect the invention, the sample isolated in step (a) is peripheral blood, and even more preferably they are mononuclear cells obtained from peripheral blood.

Steps (a), (b), and of the methods described above can be totally or partially automated, for example, by means of a robotic sensor device for the quantification of step (a) or the computerized calculation of any of the indices of steps (b), (c) and / or (d), or the computerized classification in the steps

A fourth aspect of the invention relates to a kit or device for the diagnosis and classification of a disease that develops with episodic vestibular syndrome, hereafter referred to as the kit or device of the invention, which comprises the elements necessary to detect the levels of Proinflammatory cytokines IL1β, IL6 and TNFα in an isolated biological sample of an individual. In a preferred embodiment of this aspect the invention, the isolated sample is peripheral blood, and even more preferably they are mononuclear cells obtained from peripheral blood. In another preferred embodiment of this aspect, the kit or device of the invention comprises primers, probes and / or antibodies capable of detecting and quantifying the proinflammatory cytokines IL1β, IL6 and TNFα in an isolated biological sample. Even more preferably, the detection of proinflammatory cytokine levels is performed by immunological techniques, with antibodies, and even more preferably by ELISA. In another preferred embodiment of this aspect of the invention, the antibodies are modified. In another preferred embodiment of this aspect of the invention, the antibody is human, humanized or synthetic. In another preferred embodiment, the antibody is monoclonal and / or is labeled with a fluorochrome. Preferably, the fluorochrome is selected from the list comprising Fluorescein (FITC), Tetramethylrodamine and derivatives, Phycoerythrin (PE), PerCP, Cy5, Texas, allophycocyanin, or any combination thereof.

In another preferred embodiment of this aspect of the invention, the kit or device of the invention is a kit of parts, comprising a component A, formed by a device

for the collection of the sample from step a), and a component B, formed by the elements necessary to carry out the quantitative analysis in the sample from step a) or any of the methods of the invention.

A fifth aspect of the invention relates to the use of the kit of the invention, for the diagnosis and classification of a disease that occurs with episodic vestibular syndrome. In a preferred embodiment of this aspect of the invention, the disease that occurs with vestibular syndrome is MS or MV, being used for the differential diagnosis of MS and MV, and more specifically, for the diagnosis of autoimmune MS.

BRIEF DESCRIPTION OF THE FIGURES

Fig. 1. Interleukin 1β levels in patients with Meniere's disease and controls obtained after the in vitro mononuclear cell test Fig. 2. Interleukin 6 levels in patients with Meniere's disease and controls obtained after the in vitro mononuclear cell test Fig. 3. TNFα levels in patients with Meniere's disease and controls obtained after the in vitro mononuclear cell assay. Fig. 4. Analysis of hierarchical clusters that allows patients to be classified into 2 groups according to their cytokine levels.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides a method of determining the concentration of proinflammatory cytokines for the differential diagnosis of vestibular migraine and Meniere's disease. The clinical picture is characterized by episodic vestibular symptoms associated with migrainous symptoms, including migraine headache, sensory aura, sensorineural hearing loss, tinnitus and vertigo crisis. The authors of the present invention are the first to demonstrate a relationship between proinflammatory cytokine levels in an in vitro assay using peripheral blood mononuclear cells, which allows the vestibular migraine to be differentiated from Meniere's disease.

Therefore, a first aspect of the invention relates to the use of the proinflammatory cytokines IL1β, IL6, TNFα or any combination thereof to obtain useful data for the diagnosis and classification of a disease that develops with episodic vestibular syndrome. In a preferred embodiment, the proinflammatory cytokines IL1β, IL6 and

TNFα are used simultaneously. In another preferred embodiment, the determination of proinflammatory cytokines is performed in vitro in human mononuclear cells. In another even more preferred embodiment, the classification of the disease with vestibular syndrome allows obtaining useful data for the differential diagnosis between MS and

5 MV, and more specifically, for the diagnosis of autoimmune MS.

MS may present with migrainous symptoms, including headache and MV may have sensorineural hearing loss, being indistinguishable in the early stages of the disease.

A second aspect of the invention relates to a method of obtaining useful data for the diagnosis and classification of a disease that develops with episodic vestibular syndrome, hereafter referred to as the first method of the invention, comprising:

a) detect the levels of the proinflammatory cytokines IL1β, IL6, TNFα, or any combination thereof, in an isolated biological sample of an individual. b) compare the levels of proinflammatory cytokines detected in step (a), with baseline levels in a healthy individual.

In another preferred embodiment the sample isolated in step (a) is peripheral blood, and even more preferably they are mononuclear cells obtained from peripheral blood.

In another even more preferred embodiment, the classification of the disease with vestibular syndrome allows obtaining useful data for the differential diagnosis between MS and MV, and more specifically, for the diagnosis of autoimmune MS.

That is, step (b) consists of detecting the concentrations of IL1β, IL6 or TNFα cytokines or any of their combinations. Preferably, all cytokines of the diagnostic panel of the invention are detected. Simultaneously it means that all are detected to carry out the method of the invention, although they can be detected in any order, without

30 time limit.

An "isolated biological sample" includes, but is not limited to, cells, tissues and / or biological fluids of an organism, obtained by any method known to a person skilled in the art. Preferably, the biological sample isolated from step (a) is a sample of

35 peripheral blood.

The isolated biological sample is peripheral blood, and / or comprises peripheral blood cells (PBCs), more preferably mononuclear cells. These cells are isolated, for example but not limited to, by density gradient separation using Ficoll, which is a density gradient medium based on the principle of differential migration of the blood cells through the media during the centrifugation stage of the process. Once isolated, these cells are incubated under controlled conditions, which make subsequent determinations reliable.

Basal concentrations of the cytokines IL1β, IL6 and TNFα are obtained after maintaining the mononuclear cells for 16 hours in RPMI1640 medium supplemented at conditions of 37 ° C and 7% CO2.

IL-1β - Interleukin-1β is a member of the cytokine family 1. This is produced through macrophages activated as pro-protein, which by proteolytic processes is processed to its active form thanks to caspase 1 (CASP1). This protein is a very important mediator in the inflammatory response, and is involved in a wide variety of cellular activities, including cell proliferation, differentiation and apoptosis. The increase in the production of this cytokine is observed in different diseases such as autoimmune disease of the inner ear, Muckle Wells syndrome or systemic inflammatory disease of neonatal onset.

IL-6 - Interleukin-6 is a glycoprotein secreted by macrophages, T cells and endothelial cells to stimulate the immune response. It also helps in the maturation of B cells and is an antagonist of regulatory T cells. It is a protein with both inflammatory and anti-inflammatory capacity. This cytokine is first produced at sites of acute or chronic inflammation, where it is secreted to the plasma and induces an inflammatory response. It is an important mediator of fever and acute response. IL-6 stimulates inflammatory and autoimmune processes in various diseases, such as diabetes, lupus or rheumatoid arthritis.

TNFα - Tumor necrosis factor alpha is a proinflammatory cytokine that belongs to the tumor necrosis factor superfamily. It is mostly secreted by macrophages and is involved in a broad spectrum of biological processes that include lipid metabolism, coagulation, proliferation, differentiation and cell apoptosis. This cytokine is involved in a wide variety of diseases such as cancer, insulin resistance and

autoimmune diseases (rheumatoid arthritis, ankylosing spondylitis, Crohn's disease, psoriasis).

In the context of the present invention, IL-1β, IL-6 and TNFα are also defined, but not limited to, by a nucleotide or polynucleotide sequence, which constitutes the coding sequence of the sequences collected respectively in the SEQ IDs collected in the Table 3, and which would comprise various variants from: a) nucleic acid molecules encoding said cytokines comprising the nucleotide sequence of SEQ ID listed in Table 1, b) nucleic acid molecules whose hybrid complementary chain with the sequence polynucleotide of a), c) nucleic acid molecules whose sequence differs from a) and / or b) due to the degeneracy of the genetic code, d) nucleic acid molecules encoding said cytokines comprising the nucleotide sequence with an identity of at least 80%, 90%, 95%, 98% or 99% with the SEQ IDs listed in Table 1, respectively, and in which the cytokines encoded by said acid The nuclei possesses the activity and structural characteristics of the cytokines IL-1β, IL-6 and TNFα. Among these nucleic acid molecules are those collected in the SEQ IDs indicated in table 1.

Sequences of the invention: SEQ ID NO: 1

GTGTCTGAAGCAGCCATGGCAGAAGTACCTGAGCTCGCCAGTGAAATGATGGCTTATTA CAGTGGCAATGAGGATGACTTGTTCTTTGAAGCTGATGGCCCTAAACAGATGAAGTGCT CCTTCCAGGACCTGGACCTCTGCCCTCTGGATGGCGGCATCCAGCTACGAATCTCCGA CCACCACTACAGCAAGGGCTTCAGGCAGGCCGCGTCAGTTGTTGTGGCCATGGACAAG CTGAGGAAGATGCTGGTTCCCTGCCCACAGACCTTCCAGGAGAATGACCTGAGCACCT TCTTTCCCTTCATCTTTGAAGAAGGTAGTTAGCCAAGAGCAGGCAGTAGATCTCCACTTG TGTCCTCTTGGAAGTCATCAAGCCCCAGCCAACTCAATTCCCCCAGAGCCAAAGCCCTT TAAAGGTAGAAGGCCCAGCGGGGAGACAAAACAAAGAAGGCTGGAAACCAAAGCAATC ATCTCTTTAGTGGAAACTATTCTTAAAGAAGATCTTGATGGCTACTGACATTTGCAACTC CCTCACTCTTTCTCAGGGGCCTTTCACTTACATTGTCACCAGAGAACCTATCTTCTTCGA CACATGGGATAACGAGGCTTATGTGCACGATGCACCTGTACGATCACTGAACTGCACGC TCCGGGACTCACAGCAAAAAAGCTTGGTGATGTCTGGTCCATATGAACTGAAAGCTCTC CACCTCCAGGGACAGGATATGGAGCAACAAGTGGTGTTCTCCATGTCCTTTGTACAAGG

AGAAGAAAGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGAAAAGAATCTGTACC TGTCCTGCGTGTTGAAAGATGATAAGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAA AATTACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCAACAAGATAGAAATCAATAAC AAGCTGGAATTTGAGTCTGCCCAGTTCCCCAACTGGTACATCAGCACCTCTCAAGCAGA AAACATGCCCGTCTTCCTGGGAGGGACCAAAGGCGGCCAGGATATAACTGACTTCACC ATGCAATTTGTGTCTTCCTAAAGAGAGCTGTACCCAGAGAGTCCTGTGCTGAATGTGGA CTCAATCCCTAGGGCTGGCAGAAAGGGAACAGAAAGGTTTTTGAGTACGGCTATAGCCT GGACTTTCCTGTTGTCTACACCAATGCCCAACTGCCTGCCTTAGGGTAGTGCTAAGAGG ATCTCCTGTCCATCAGCCAGGACAGTCAGCTCTCTCCTTTCAGGGCCAATCCCCAGCCC TTTTGTTGAGCCAGGCCTCTCTCACCTCTCCTACTCACTTAAAGCCCGCCTGACAGAAA CCACGGCCACATTTGGTTCTAAGAAACCCTCTGTCATTCGCTCCCACATTCTGATGAGC AACCGCTTCCCTATTTATTTATTTATTTGTTTGTTTGTTTTATTCATTGGTCTAATTTATTCA AAGGGGGCAAGAAGTAGCAGTGTCTGTAAAAGAGCCTAGTTTTTAATAGCTATGGAATC AATTCAATTTGGACTGGTGTGCTCTCTTTAAATCAAGTCCTTTAATTAAGACTGAAAATAT ATAAGCTCAGATTATTTAAATGGGAATATTTATAAATGAGCAAATATCATACTGTTCAATG GTTCTGAAATAAACTTCACTGAAGAAAAAAAAA

SEQ ID NO: 2 MATDICNSLTLSQGPFTYIVTREPIFFDTWDNEAYVHDAPVRSLNCTLRDSQQKSLVMSGPY ELKALHLQGQDMEQQVVFSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTLQLESV DPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPNWYISTSQAENMPVFLGGTKGGQDITDFT MQFVSS

SEQ ID NO: 3 ACCAAACCTCTTCGAGGCACAAGGCACAACAGGCTGCTCTGGGATTCTCTTCAGCCAAT CTTCATTGCTCAAGTGTCTGAAGCAGCCATGGCAGAAGTACCTGAGCTCGCCAGTGAAA TGATGGCTTATTACAGTGGCAATGAGGATGACTTGTTCTTTGAAGCTGATGGCCCTAAA CAGATGAAGTGCTCCTTCCAGGACCTGGACCTCTGCCCTCTGGATGGCGGCATCCAGC TACGAATCTCCGACCACCACTACAGCAAGGGCTTCAGGCAGGCCGCGTCAGTTGTTGT GGCCATGGACAAGCTGAGGAAGATGCTGGTTCCCTGCCCACAGACCTTCCAGGAGAAT GACCTGAGCACCTTCTTTCCCTTCATCTTTGAAGAAGAACCTATCTTCTTCGACACATGG GATAACGAGGCTTATGTGCACGATGCACCTGTACGATCACTGAACTGCACGCTCCGGG ACTCACAGCAAAAAAGCTTGGTGATGTCTGGTCCATATGAACTGAAAGCTCTCCACCTC CAGGGACAGGATATGGAGCAACAAGTGGTGTTCTCCATGTCCTTTGTACAAGGAGAAGA AAGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGAAAAGAATCTGTACCTGTCCT GCGTGTTGAAAGATGATAAGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAAAATTAC

CCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCAACAAGATAGAAATCAATAACAAGCT GGAATTTGAGTCTGCCCAGTTCCCCAACTGGTACATCAGCACCTCTCAAGCAGAAAACA TGCCCGTCTTCCTGGGAGGGACCAAAGGCGGCCAGGATATAACTGACTTCACCATGCA ATTTGTGTCTTCCTAAAGAGAGCTGTACCCAGAGAGTCCTGTGCTGAATGTGGACTCAA TCCCTAGGGCTGGCAGAAAGGGAACAGAAAGGTTTTTGAGTACGGCTATAGCCTGGAC TTTCCTGTTGTCTACACCAATGCCCAACTGCCTGCCTTAGGGTAGTGCTAAGAGGATCT CCTGTCCATCAGCCAGGACAGTCAGCTCTCTCCTTTCAGGGCCAATCCCCAGCCCTTTT GTTGAGCCAGGCCTCTCTCACCTCTCCTACTCACTTAAAGCCCGCCTGACAGAAACCAC GGCCACATTTGGTTCTAAGAAACCCTCTGTCATTCGCTCCCACATTCTGATGAGCAACC GCTTCCCTATTTATTTATTTATTTGTTTGTTTGTTTTATTCATTGGTCTAATTTATTCAAAG GGGGCAAGAAGTAGCAGTGTCTGTAAAAGAGCCTAGTTTTTAATAGCTATGGAATCAATT CAATTTGGACTGGTGTGCTCTCTTTAAATCAAGTCCTTTAATTAAGACTGAAAATATATAA GCTCAGATTATTTAAATGGGAATATTTATAAATGAGCAAATATCATACTGTTCAATGGTTC TGAAATAAACTTCACTGAAG

SEQ ID NO: 4 MAEVPELASEMMAYYSGNEDDLFFEADGPKQMKCSFQDLDLCPLDGGIQLRISDHHYSKG FRQAASVVVAMDKLRKMLVPCPQTFQENDLSTFFPFIFEEEPIFFDTWDNEAYVHDAPVRSL NCTLRDSQQKSLVMSGPYELKALHLQGQDMEQQVVFSMSFVQGEESNDKIPVALGLKEKN LYLSCVLKDDKPTLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPNWYISTSQAE NMPVFLGGTKGGQDITDFTMQFVSS

SEQ ID NO: 5 GTATTGTGTCACTCAGTTCAAGTACTTGAAATTTATTGAATTGTATTTTCTAAAAAATAGAT AGTTGAGTAAAAGCAAGCTCACATTACATAGACGGATCACAGTGCACGGCTGCGGAGCT GGGAGCAGTGGCTTCGTTTCATGCAGGAAAGAGAACTTGGTTCAGGAGTGTCTACGTT GCTTAAGACAGGAGAGCACTAAAAATGAAACCATCCAGCCATCCTCCCCCATTTTCATTT TCACACCAAAGAATCCCACCGCGGCAGAGGACCACCGTCTCTGTTTAGACAATCGGTG AAGAATGGATGACCTCACTTTCCCCAACAGGCGGGTCCTGAAATGTTATGCACGAAACA AAACTTGAGTAAATGCCCAACAGAGGTCACTGTTTTATCGATCTTGAAGAGATCTCTTCT TAGCAAAGCAAAGAAACCGATTGTGAAGGTAACACCATGTTTGGTAAATAAGTGTTTTGG TGTTGTGCAAGGGTCTGGTTTCAGCCTGAAGCCATCTCAGAGCTGTCTGGGTCTCTGGA GACTGGAGGGACAACCTAGTCTAGAGCCCATTTGCATGAGACCAAGGATCCTCCTGCA AGAGACACCATCCTGAGGGAAGAGGGCTTCTGAACCAGCTTGACCCAATAAGAAATTCT TGGGTGCCGACGCGGAAGCAGATTCAGAGCCTAGAGCCGTGCCTGCGTCCGTAGTTTC CTTCTAGCTTCTTTTGATTTCAAATCAAGACTTACAGGGAGAGGGAGCGATAAACACAAA

CTCTGCAAGATGCCACAAGGTCCTCCTTTGACATCCCCAACAAAGAGGACTGGAGATGT CTGAGGCTCATTCTGCCCTCGAGCCCACCGGGAACGAAAGAGAAGCTCTATCTCCCCT CCAGGAGCCCAGCTATGAACTCCTTCTCCACAAGCGCCTTCGGTCCAGTTGCCTTCTCC CTGGGGCTGCTCCTGGTGTTGCCTGCTGCCTTCCCTGCCCCAGTACCCCCAGGAGAAG ATTCCAAAGATGTAGCCGCCCCACACAGACAGCCACTCACCTCTTCAGAACGAATTGAC AAACAAATTCGGTACATCCTCGACGGCATCTCAGCCCTGAGAAAGGAGACATGTAACAA GAGTAACATGTGTGAAAGCAGCAAAGAGGCACTGGCAGAAAACAACCTGAACCTTCCAA AGATGGCTGAAAAAGATGGATGCTTCCAATCTGGATTCAATGAGGAGACTTGCCTGGTG AAAATCATCACTGGTCTTTTGGAGTTTGAGGTATACCTAGAGTACCTCCAGAACAGATTT GAGAGTAGTGAGGAACAAGCCAGAGCTGTGCAGATGAGTACAAAAGTCCTGATCCAGT TCCTGCAGAAAAAGGCAAAGAATCTAGATGCAATAACCACCCCTGACCCAACCACAAAT GCCAGCCTGCTGACGAAGCTGCAGGCACAGAACCAGTGGCTGCAGGACATGACAACTC ATCTCATTCTGCGCAGCTTTAAGGAGTTCCTGCAGTCCAGCCTGAGGGCTCTTCGGCAA ATGTAGCATGGGCACCTCAGATTGTTGTTGTTAATGGGCATTCCTTCTTCTGGTCAGAAA CCTGTCCACTGGGCACAGAACTTATGTTGTTCTCTATGGAGAACTAAAAGTATGAGCGTT AGGACACTATTTTAATTATTTTTAATTTATTAATATTTAAATATGTGAAGCTGAGTTAATTT ATGTAAGTCATATTTATATTTTTAAGAAGTACCACTTG AAACATTTTATGTATTAGTTTTGA AATAATAATGGAAAGTGGCTATGCAGTTTGAATATCCTTTGTTTCAGAGCCAGATCATTT CTTGGAAAGTGTAGGCTTACCTCAAATAAATGGCTAACTTATACATATTTTTAAAGAAATATATATATAATTATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATAATTATATATAATTATTATATATAATTATATATAATTATATATATA

SEQ ID NO: 6 MNSFSTSAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDG ISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYL EYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQD MTTHLILRSFKEFLQSSLRALRQM

SEQ ID NO: 7 CCCGCTCTGGCCCCACCCTCACCCTCCAACAAAGATTTATCAAATGTGGGATTTTCCCA TGAGTCTCAATATTAGAGTCTCAACCCCCAATAAATATAGGACTGGAGATGTCTGAGGCT CATTCTGCCCTCGAGCCCACCGGGAACGAAAGAGAAGCTCTATCTCCCCTCCAGGAGC CCAGCTATGAACTCCTTCTCCACAAGTAAGTGCAGGAAATCCTTAGCCCTGGAACTGCC AGCGGCGGTCGAGCCCTGTGTGAGGGAGGGGTGTGTGGCCCAGGGAGGGCTGGCGG GCGGCCAGCAGCAGAGGCAGGCTCCCAGCTGTGCTGTCAGCTCACCCCTGCGCTCGC TCCCCTCCGGCACAGGCGCCTTCGGTCCAGTTGCCTTCTCCCTGGGGCTGCTCCTGGT

GTTGCCTGCTGCCTTCCCTGCCCCAGTACCCCCAGGAGAAGATTCCAAAGATGTAGCC GCCCCACACAGACAGCCACTCACCTCTTCAGAACGAATTGACAAACAAATTCGGTACAT CCTCGACGGCATCTCAGCCCTGAGAAAGGAGACATGTAACAAGAGTAACATGTGTGAAA GCAGCAAAGAGGCACTGGCAGAAAACAACCTGAACCTTCCAAAGATGGCTGAAAAAGA TGGATGCTTCCAATCTGGATTCAATGAGGAGACTTGCCTGGTGAAAATCATCACTGGTC TTTTGGAGTTTGAGGTATACCTAGAGTACCTCCAGAACAGATTTGAGAGTAGTGAGGAA CAAGCCAGAGCTGTGCAGATGAGTACAAAAGTCCTGATCCAGTTCCTGCAGAAAAAGGT GGGTGTGTCCTCATTCCCTCAACTTGGTGTGGGGGAAGACAGGCTCAAAGACAGTGTC CTGGACAACTCAGGGATGCAATGCCACTTCCAAAAGAGAAGGCTACACGTAAACAAAAG AGTCTGAGAAATAGTTTCTGATTGTTATTGTTAAATCTTTTTTTGTTTGTTTGGTTGGTTG GCTCTCTTCTGCAAAGGACATCAATAACTGTATTTTAAACTATATATTAACTGAGGTGGAT TTTAACATCAATTTTTAATAGTGCAAGAGATTTAAAACCAAAGGCGGGGGGGCGGGCAG AAAAAAGTGCATCCAACTCCAGCCAGTGATCCACAGAAACAAAGACCAAGGAGCACAAA ATGATTTTAAGATTTTAGTCATTGCCAAGTGACATTCTTCTCACTGTGGTTGTTTCAATTC TTTTTCCTACCTTTTACCAGAGAGTTAGTTCAGAGAAATGGTCAGAGACTCAAGGGTGGA AAGAGGTACCAAAGGCTTTGGCCACCAGTAGCTGGCTATTCAGACAGCAGGGAGTAGA CTTGCTGGCTAGCATGTGGAGGAGCCAAAGCTCAATA AGAAGGGGCCTAGAATGAAAC CCTTGGTGCTGATCCTGCCTCTGCCATTTCTACTTAAGCCAGGGTTTCTCATATGTTAAC ATGCATGGGAATTCCCTGGGCATCTTCTTGTGGTGTGGAGTCTGACTTAGCAAGCCTCG GGTGGGTTTGAGGGTCAAATTTCTACCAGGCTTATATCCCTGGTGATGCTGCAGAATTC CAGGACCACACTTGGAGGTTTAAGGCCTTCCACAAGTTACTTATCCCATATGGTGGGTC TATGGAAAGGTGTTTCCCAGTCCTCTTTACACCACCGGATCAGTGGTCTTTCAACAGATC CTAAAGGGATGGTGAGAGGGAAACTGGAGAAAAGTATCAGATTTAGAGGCCACTGAAG AACCCATATTAAAATGCCTTTAAGTATGGGCTCTTCATTCATATACTAAATATGAACTATG TGCCAGGCATTATTTCATATGACAGAATACAAACAAATAAGATAGTGATGCTGGTCAGGC TTGGTGGCTCATGCCTGTATTCCCTAAACTTTGGGAGCCTAAGGTGAGAACTCCTTGAA CTCCTAAGGCCAGGAGTTCAAGACCAGCCTGGATAACATAGCAAGACCCCATCTCTACA AAAAACCAAAACCAAACAAACAAAAATGATAGTGGTGCTTCCCTCAGGATGCTTGTGGT CTAATGGGAGACAGAACAGCAAAGGGATGATTAGAAGTTGGTTGCTGTGAGCCAGGCA CAGTGCTGATATAATCCCAGCGCTATGGGAGGCTGAGGTGGGTGGATCATTTGAGGCC AGGAGTTTAAGACCAGCCTGGTCAACATGGTAAAACCCCATCTCTACTTAAAAATACAAA AAAGTTAGCCAGGCATGGTGGCATACACCTGTAACCCAGCTACTCAGGAGGCTGAGGC ACATGAATCACTTGAACCCAGGAGGCAGAGGTTGCTGTGCACCACTGCACTCCAGCCT GGGTGACAGAACGAGAC CTTGACTCAAAAAAAAAAAAAAGAAGTTTGTTGCTATGGAAG GGTCCTACTCAGAGCAGGCACCCCAGTTAATCTCATTCACCCCACATTTCACATTTGAAC

ATCATCCCATAGCCCAGAGCATCCCTCCACTGCAAAGGATTTATTCAACATTTAAACAAT CCTTTTTACTTTCATTTTC

SEQ ID NO: 8 MNSFSTSKCRKSLALELPAAVEPCVREGCVAQGGLAGGQQQRQAPSCAVSSPLRSLPSGT GAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDGISALRK ETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQN RFESSEEQARAVQMSTKVLIQFLQKKVGVSSFPQLGVGEDRLKDSVLDNSGMQCHFQKRR LHVNKRV

SEQ ID NO: 9 AATATTAGAGTCTCAACCCCCAATAAATATAGGACTGGAGATGTCTGAGGCTCATTCTGC CCTCGAGCCCACCGGGAACGAAAGAGAAGCTCTATCTCCCCTCCAGGAGCCCAGCTAT GAACTCCTTCTCCACAAGCGCCTTCGGTCCAGTTGCCTTCTCCCTGGGGCTGCTCCTG GTGTTGCCTGCTGCCTTCCCTGCCCCAGTACCCCCAGGAGAAGATTCCAAAGATGTAG CCGCCCCACACAGACAGCCACTCACCTCTTCAGAACGAATTGACAAACAAATTCGGTAC ATCCTCGACGGCATCTCAGCCCTGAGAAAGGAGACATGTAACAAGAGTAACATGTGTGA AAGCAGCAAAGAGGCACTGGCAGAAAACAACCTGAACCTTCCAAAGATGGCTGAAAAA GATGGATGCTTCCAATCTGGATTCAATGAGGAGACTTGCCTGGTGAAAATCATCACTGG TCTTTTGGAGTTTGAGGTATACCTAGAGTACCTCCAGAACAGATTTGAGAGTAGTGAGG AACAAGCCAGAGCTGTGCAGATGAGTACAAAAGTCCTGATCCAGTTCCTGCAGAAAAAG GCAAAGAATCTAGATGCAATAACCACCCCTGACCCAACCACAAATGCCAGCCTGCTGAC GAAGCTGCAGGCACAGAACCAGTGGCTGCAGGACATGACAACTCATCTCATTCTGCGC AGCTTTAAGGAGTTCCTGCAGTCCAGCCTGAGGGCTCTTCGGCAAATGTAGCATGGGC ACCTCAGATTGTTGTTGTTAATGGGCATTCCTTCTTCTGGTCAGAAACCTGTCCACTGGG CACAGAACTTATGTTGTTCTCTATGGAGAACTAAAAGTATGAGCGTTAGGACACTATTTT AATTATTTTTAATTTATTAATATTTAAATATGTGAAGCTGAGTTAATTTATGTAAGTCATATT TATATTTTTAAGAAGTACCACTTGAAA CATTTTATGTATTAGTTTTGAAATAATAATGGAAA GTGGCTATGCAGTTTGAATATCCTTTGTTTCAGAGCCAGATCATTTCTTGGAAAGTGTAG GCTTACCTCAAATAAATGGCTAACTTATACATATTTTTAAAGAAATATTTATATTGTATTTA TATAATGTATAAATGGTTTTTATACCAATAAATGGCATTTTAAAAAATTCAGCAAAAAAAAA AAAAAAAAAA

SEQ ID NO: 10 MNSFSTSAFGPVAFSLGLLLVLPAAFPAPVPPGEDSKDVAAPHRQPLTSSERIDKQIRYILDG ISALRKETCNKSNMCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYL

EYLQNRFESSEEQARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQD MTTHLILRSFKEFLQSSLRALRQM

SEQ ID NO: 11 GTCTCAATATTAGAGTCTCAACCCCCAATAAATATAGGACTGGAGATGTCTGAGGCTCAT TCTGCCCTCGAGCCCACCGGGAACGAAAGAGAAGCTCTATCTCCCCTCCAGGAGCCCA GCTATGAACTCCTTCTCCACAAACATGTAACAAGAGTAACATGTGTGAAAGCAGCAAAG AGGCACTGGCAGAAAACAACCTGAACCTTCCAAAGATGGCTGAAAAAGATGGATGCTTC CAATCTGGATTCAATGAGGAGACTTGCCTGGTGAAAATCATCACTGGTCTTTTGGAGTTT GAGGTATACCTAGAGTACCTCCAGAACAGATTTGAGAGTAGTGAGGAACAAGCCAGAG CTGTGCAGATGAGTACAAAAGTCCTGATCCAGTTCCTGCAGAAAAAGGCAAAGAATCTA GATGCAATAACCACCCCTGACCCAACCACAAATGCCAGCCTGCTGACGAAGCTGCAGG CACAGAACCAGTGGCTGCAGGACATGACAACTCATCTCATTCTGCGCAGCTTTAAGGAG TTCCTGCAGTCCAGCCTGAGGGCTCTTCGGCAAATGTAGCATGGGCACCTCAGATTGTT GTTGTTAATGGGCATTCCTTCTTCTGGTCAGAAACCTGTCCACTGGGCACAGAACTTAT GTTGTTCTCTATGGAGAACTAAAAGTATGAGCGTTAGGACACTATTTTAATTATTTTTAAT TTATTAATATTTAAATATGTGAAGCTGAGTTAATTTATGTAAGTCATATTTATATTTTTAAG AAGTACCACTTGAAACATTTTATGTATTAGTTTTGAAATAATAATGGAAAGTGGCTATGCA GTTTGAATATCCTTTGTTTCAGAGCCAGATCATTTCTTGGAAAGTGTAGGCTTACCTCAA ATAAATGGCTAACTTATACATATTTTTAAAGAAATATTTATATTGTATTTATATAATGTATAA ATGGTTTTTATACCAA TAAATGGCATTTTAAAAAATTCAGCAAAAAAAAAA

SEQ ID NO: 12 MCESSKEALAENNLNLPKMAEKDGCFQSGFNEETCLVKIITGLLEFEVYLEYLQNRFESSEE QARAVQMSTKVLIQFLQKKAKNLDAITTPDPTTNASLLTKLQAQNQWLQDMTTHLILRSFKE FLQQLSL

SEQ ID NO: 13 CAGACGCTCCCTCAGCAAGGACAGCAGAGGACCAGCTAAGAGGGAGAGAAGCAACTA CAGACCCCCCCTGAAAACAACCCTCAGACGCCACATCCCCTGACAAGCTGCCAGGCAG GTTCTCTTCCTCTCACATACTGACCCACGGCTCCACCCTCTCTCCCCTGGAAAGGACAC CATGAGCACTGAAAGCATGATCCGGGACGTGGAGCTGGCCGAGGAGGCGCTCCCCAA GAAGACAGGGGGGCCCCAGGGCTCCAGGCGGTGCTTGTTCCTCAGCCTCTTCTCCTTC CTGATCGTGGCAGGCGCCACCACGCTCTTCTGCCTGCTGCACTTTGGAGTGATCGGCC CCCAGAGGGAAGAGTTCCCCAGGGACCTCTCTCTAATCAGCCCTCTGGCCCAGGCAGT CAGATCATCTTCTCGAACCCCGAGTGACAAGCCTGTAGCCCATGTTGTAGCAAACCCTC

AAGCTGAGGGGCAGCTCCAGTGGCTGAACCGCCGGGCCAATGCCCTCCTGGCCAATG GCGTGGAGCTGAGAGATAACCAGCTGGTGGTGCCATCAGAGGGCCTGTACCTCATCTA CTCCCAGGTCCTCTTCAAGGGCCAAGGCTGCCCCTCCACCCATGTGCTCCTCACCCAC ACCATCAGCCGCATCGCCGTCTCCTACCAGACCAAGGTCAACCTCCTCTCTGCCATCAA

5 GAGCCCCTGCCAGAGGGAGACCCCAGAGGGGGCTGAGGCCAAGCCCTGGTATGAGCC CATCTATCTGGGAGGGGTCTTCCAGCTGGAGAAGGGTGACCGACTCAGCGCTGAGATC AATCGGCCCGACTATCTCGACTTTGCCGAGTCTGGGCAGGTCTACTTTGGGATCATTGC CCTGTGAGGAGGACGAACATCCAACCTTCCCAAACGCCTCCCCTGCCCCAATCCCTTTA TTACCCCCTCCTTCAGACACCCTCAACCTCTTCTGGCTCAAAAAGAGAATTGGGGGCTT

10 AGGGTCGGAACCCAAGCTTAGAACTTTAAGCAACAAGACCACCACTTCGAAACCTGGGA TTCAGGAATGTGTGGCCTGCACAGTGAAGTGCTGGCAACCACTAAGAATTCAAACTGGG GCCTCCAGAACTCACTGGGGCCTACAGCTTTGATCCCTGACATCTGGAATCTGGAGACC AGGGAGCCTTTGGTTCTGGCCAGAATGCTGCAGGACTTGAGAAGACCTCACCTAGAAAT TGACACAAGTGGACCTTAGGCCTTCCTCTCTCCAGATGTTTCCAGACTTCCTTGAGACA

15 CGGAGCCCAGCCCTCCCCATGGAGCCAGCTCCCTCTATTTATGTTTGCACTTGTGATTA TTTATTATTTATTTATTATTTATTTATTTACAGATGAATGTATTTATTTGGGAGACCGGGGT ATCCTGGGGGACCCAATGTAGGAGCTGCCTTGGCTCAGACATGTTTTCCGTGAAAACG GAGCTGAACAATAGGCTGTTCCCATGTAGCCCCCTGGCCTCTGTGCCTTCTTTTGATTA TGTTTTTTAAAATATTTATCTGATTAAGTTGTCTAAACAATGCTGATTTGGTGACCAACTG

20 TCACTCATTGCTGAGCCTCTGCTCCCCAGGGGAGTTGTGTCTGTAATCGCCCTACTATT CAGTGGCGAGAAATAAAGTTTGCTTAGAAAAGAAAAAAAAAAAAA

SEQ ID NO: 14

MSTESMIRDVELAEEALPKKTGGPQGSRRCLFLSLFSFLIVAGATTLFCLLHFGVIGPQREEF

25 PRDLSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANALLANGVELRDNQL VVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKVNLLSAIKSPCQRETPEGAEA KPGYEPIYLGLDGYGYGYQYGYGYQYGYGYGYGYGDGLWGDGLWGDWG

Table 3. Sequences of the invention

Gene Name

Description Gen ID Nucleotide sequences Amino acid sequences

IL1B

1 beta interleukin 3553 SEQ ID NO: 1 or SEQ ID NO: 3 SEQ ID NO: 2 or SEQ ID NO: 4

IL6

Interleukin 6 3569 SEQ ID NO: SEQ ID NO: 6,

5, SEQ ID NO: 7, SEQ ID NO: 9 or SEQ ID NO: 11

SEQ ID NO: 8, SEQ ID NO: 10 or SEQ ID NO: 12

TNF alpha

Tumor necrosis factor 7124 SEQ ID NO: 13 SEQ ID NO: 14

In another preferred embodiment of this aspect of the invention, the disease that occurs with the course of vestibular syndrome is Meniere's disease (MS) or autoimmune disease of the inner ear.

In this report it is understood as a disease that presents with episodic vestibular syndrome that in which the patient presents recurrent signs and symptoms that indicate an alteration of the vestibular function, such as vertigo, ataxia, imbalance, nystagmus or oscilopsia. These symptoms may be associated with various hearing symptoms or headache.

10 origin.

In this report, Ménière's disease is understood as the syndrome characterized by uni or bilateral sensorineural hearing loss that affects low and medium frequencies (<2000Hz) and associated with episodes of vertigo and auditory symptoms, such as tinnitus, or

15 otical fullness. In this report, autoimmune disease of the inner ear or autoimmune MS is understood as the syndrome characterized by uni or bilateral sensorineural hearing loss greater than 30dB that affects one or more frequencies and that progresses in a period between 3-90 days. The progression of hearing loss should be greater than 15dB for a frequency or 10dB for 2 frequencies and

20 demonstrated by audiometry. Autoimmune disease of the inner ear is associated with vestibular symptoms in 50% of cases. The onset of sudden hearing loss (<3 days) with an autoimmune risk profile should make autoimmune disease of the inner ear suspect.

As shown in the examples of the present invention, proinflammatory cytokine levels are elevated in a subset of patients with MS (Table 1).

Table 1. Proinflammatory cytokine levels used for the diagnosis of autoimmune Meniere's disease.

Table 2. Cytokine levels in the in vitro assay with mononuclear cells of patients with vestibular migraine and controls

10 Thus, a third aspect of the invention relates to a method for the diagnosis and classification of a disease that develops with episodic vestibular syndrome, hereafter referred to as the second method of the invention, which comprises steps (a) and (b ) of the first method of the invention, and further comprises:

15 c) include the individual in the group of individuals with MS of autoimmune origin.

In a preferred embodiment of this aspect the invention, the sample isolated in step (a) is peripheral blood, and even more preferably they are mononuclear cells obtained from peripheral blood.

The term "individual" or "subject", as used in the description, refers to an animal, preferably a mammal, and more preferably a human being. The term "individual" in this report is synonymous with "patient", and is not intended to be limiting in

no aspect, being able to be this of any age, sex and physical condition. The individual can be anyone, an individual predisposed to a disease (for example, lung cancer) or an individual suffering from said disease. . The cytokine determination of the invention can be done by any method known in the state of the art, for example, but not limited to, by techniques based on the antigen-antibody interaction such as immunoassays (radioimmunoassay, enzyme immunoassay, fluoroimmunoassay, immunochemiluminescent assay), western blot, immunoprecipitation, immunohistochemistry or immunofluorescence, or by means of quantification techniques of genetic material (RNA) such as PCR, real-time PCR, or expression assays.

The term "diagnosis", as used in the present invention, at the greater or lesser risk of suffering from Meniere's disease or at the greater or lesser risk of suffering from the autoimmune disease of the inner ear.

In turn, according to the method of the present invention, other subclassifications could be established within this principal, thus facilitating the choice and establishment of appropriate therapeutic regimens. This discrimination, as understood by one skilled in the art, is not intended to be correct in 100% of the samples analyzed. However, it requires that a statistically significant amount of the analyzed samples be classified correctly. The amount that is statistically significant can be established by a person skilled in the art by using different statistical tools, for example, but not limited, by determining confidence intervals, determining the significance value P, Student test or discriminant functions. Fisher, non-parametric measurements of Mann Whitney, Spearman correlation, logistic regression, linear regression, area under the ROC curve (AUC). Preferably, the confidence intervals are at least 90%, at least 95%, at least 97%, at least 98% or at least 99%. Preferably, the value of p is less than 0.1, 0.05, 0.01, 0.005 or 0.0001. Preferably, the present invention makes it possible to correctly detect the predisposition to the disease or disease differentially by at least 60%, more preferably at least 70%, much more preferably at least 80%, or even much more preferably in at least 90% of the subjects of a certain group or population analyzed.

Steps (a), (b), and of the methods described above may be wholly or partially

automated, for example, by means of a robotic sensor device for the quantification of step (a) or computerized calculation of any of the indices of steps (b), (c) and / or (d), or computerized classification in the steps

5 MEDICAL USES

A fourth aspect of the invention relates to the use of an immunosuppressant for the treatment of an individual from the group of individuals with MS of autoimmune origin identified according to the second method of the invention. In a preferred embodiment of this

The immunosuppressants aspect is selected from the list consisting of: cytostatic agents, glucocorticoids, calcineurin enzyme inhibitors, mTOR inhibitors, antibodies, or any combination thereof.

Even more preferably, cytostatic agents are selected from Azathioprine,

15 nucleoside synthesis inhibitors such as, but not limited to, mycophenolate mofetil and mycophenolic acid, cyclophosphamide, methotrexate, or any combination thereof.

In another preferred embodiment, the glucocorticoids are selected from prednisone, fludrocortisone, triamcinolone, betamethasone, or any combination thereof.

In another preferred embodiment, the calcineurin enzyme inhibitors are selected from cyclosporine, tacrolimus, or their combination.

In another preferred embodiment, mTOR inhibitors are selected from Sirolimus, Everolimus, or their combination.

In another preferred embodiment the antibodies are selected from: anti-CD20, Rituximab; anti-IL-2R, Daclizumab, anti-TNF, Remicade, anti-SR-TNF, Enbrel, Anti-chemokines, or

30 any of their combinations.

The different treatments of the diseases that occur with episodic vestibular syndrome are described below.

35 Preventive treatment of vestibular migraine attacks

1. Anticonvulsants (Valproate, topiramate)

2.

 Antidepressants (amitriptyline, nortriptyline, venlafaxine)

3.

 Beta-blockers (propranolol, metoprolol, atenolol)

Four.

 Calcium antagonists (flunarizine)

5.

 Carbonic anhydrase inhibitor: acetazolamide The treatment is personalized based on other associated pathologies.

Preventive treatment of Meniere's disease crises

one.

 Salt-free diet and water overload (> 2000ml water / day)

2.

 Betahistine

3.

 Diuretics

Four.

 Intratimpanic gentamicin

5.

 Intratimpanic corticosteroids

6.

 Surgery (vestibular neurectomy, labytectomy according to auditory function)

Treatment of autoimmune Meniere's disease and / or autoimmune inner ear disease

one.

 Glucocorticoids (prednisone, methyl prednisolone) These are all potential treatments, which could be applied in CD:

2.

Immunosuppressants such as cytostatics (cyclophosphamide), folic acid analogs (methotrexate), purine analogs (azathioprine), immunophilin regulatory drugs (cyclosporine, tacrolimus)

3.

 Monoclonal antibodies against interleukins or their receptors such as

3.1. AntiTNFalfa: infliximab, etanercept, adalimumab.

3.2. Anti-IL-1beta receptor: Anakinra (this drug is already patented for the treatment of autoimmune disease of the inner ear, but not for the autoimmune Meniere).

3.3. Anti IL-6: Tolizizumab, siltuximab, sarilumab, olokizumab.

3.4. Anti TWEAK / FN14: BIIB023 (BIOGEN)

KIT OR DEVICE OF THE INVENTION

A fifth aspect of the invention relates to a kit or device for the diagnosis and classification of a disease that occurs with episodic vestibular syndrome, hereafter referred to as the kit or device of the invention, comprising the elements necessary to detect the levels of Proinflammatory cytokines IL1β, IL6 and TNFα in an isolated biological sample of an individual. In a preferred embodiment of this aspect the invention, the isolated sample is peripheral blood, and even more preferably they are cells

mononuclear obtained from peripheral blood. In another preferred embodiment of this aspect, the kit or device of the invention comprises primers, probes and / or antibodies capable of detecting and quantifying the proinflammatory cytokines IL1β, IL6 and TNFα in an isolated biological sample. Even more preferably, the detection of proinflammatory cytokine levels is performed by immunological techniques, with antibodies, and even more preferably by ELISA. In another preferred embodiment of this aspect of the invention, the antibodies are modified. In another preferred embodiment of this aspect of the invention, the antibody is human, humanized or synthetic. In another preferred embodiment, the antibody is monoclonal and / or is labeled with a fluorochrome. Preferably, the fluorochrome is selected from the list comprising Fluorescein (FITC), Tetramethylrodamine and derivatives, Phycoerythrin (PE), PerCP, Cy5, Texas, allophycocyanin, or any combination thereof.

Therefore, preferably, the antibodies of the kit of the invention are conjugated to molecules that emit detectable signals such as a radioactive isotope, an enzyme, a fluorescent particle or a chemiluminescent substance or assays that are measured by light scattering or direct visualization such as they are precipitation, agglutination or radial diffusion.

Said kit or device may contain all those reagents necessary to determine the cytokines of the invention by means of any of the methods existing in the state of the art and / or described herein. The kit can also include, without any limitation, buffers, agents to prevent contamination, inhibitors of protein degradation, etc. On the other hand, the kit can include all the supports and containers necessary for commissioning and optimization. Preferably, the kit further comprises instructions for carrying out any of the methods of the invention.

It is also possible that the antibody (s) are immobilized in spots on a (preferably solid) surface. In one of its embodiments, the kit comprises a microarray, or microarray of the invention. An antibody microarray is a matrix on a solid substrate (usually a glass holder or a thin silicon film cell). Detection is performed colorimetrically by the interaction of a chromogenic substrate and an enzyme that has been coupled to a detector antibody. Each stain contains the antibody (s). Although the number of spots is not limited in any way, there is a preferred embodiment in which the matrix is customized for

methods of the invention. In one embodiment, said custom matrix comprises fifty spots or less, such as thirty spots or less, including twenty spots

or less. Therefore, another aspect of the invention relates to a matrix comprising the antibody (s) of the invention.

Synthesis in situ on a solid support (for example, glass) could be done using ink-jet technology, which requires longer probes. The supports could be, but not limited to, filters or membranes of NC or nylon (charged), silicon, or glass slides for microscopes covered with aminosilanes, polylysine, aldehydes or epoxy.

In another preferred embodiment of this aspect of the invention, the kit or device of the invention is a kit of parts, comprising a component A, formed by a device for collecting the sample from step a), and a component B, formed by the elements necessary to carry out the quantitative analysis in the sample from step a) or any of the methods of the invention.

A sixth aspect of the invention relates to the use of the kit of the invention, for the diagnosis and classification of a disease that occurs with episodic vestibular syndrome. In a preferred embodiment of this aspect of the invention, the disease that occurs with vestibular syndrome is MS or MV, being used for the differential diagnosis of MS and MV, and more specifically, for the diagnosis of autoimmune MS.

The invention also extends to computer programs adapted so that any processing means can implement the methods of the invention. Such programs may have the form of source code, object code, an intermediate source of code and object code, for example, as in partially compiled form, or in any other form suitable for use in the implementation of the processes according to the invention . Computer programs also cover cloud applications based on that procedure.

Therefore, a seventh aspect of the invention relates to a computer program comprising program instructions to make a computer carry out the method according to any of the methods of the invention.

In particular, the invention encompasses computer programs arranged on or within a carrier. The carrier can be any entity or device capable of supporting the program. When the program is incorporated into a signal that can be directly transported by a cable or other device or medium, the carrier may be constituted by said cable or other device or means. As a variant, the carrier could be an integrated circuit in which the program is included, the integrated circuit being adapted to execute, or to be used in the execution of, the corresponding processes.

For example, the programs could be incorporated into a storage medium, such as a ROM, a CD ROM or a semiconductor ROM, a USB memory, or a magnetic recording medium, for example, a floppy disk or a disk Lasted. Alternatively, the programs could be supported on a transmissible carrier signal. For example, it could be an electrical or optical signal that could be transported through an electrical or optical cable, by radio or by any other means.

An eighth aspect of the invention relates to a computer-readable storage medium comprising program instructions capable of having a computer perform the steps of any of the methods of the invention.

A ninth aspect of the invention relates to a transmissible signal comprising program instructions capable of having a computer perform the steps of any of the methods of the invention.

The terms "polynucleotide" and "nucleic acid" are used interchangeably herein, referring to polymeric forms of nucleotides of any length, both ribonucleotides (RNA or RNA) and deoxyribonucleotides (DNA or DNA).

The terms "amino acid sequence", "peptide", "oligopeptide", "polypeptide" and "protein" are used interchangeably herein, and refer to a polymeric form of amino acids of any length, which may be coding or non-coding, Chemically or biochemically modified.

Throughout the description and the claims the word "comprises" and its variants are not intended to exclude other technical characteristics, additives, components or steps. For those skilled in the art, other objects, advantages and characteristics of the invention are

they will come partly from the description and partly from the practice of the invention. The following examples and drawings are provided by way of illustration, and are not intended to be limiting of the present invention.

5 EXAMPLES OF THE INVENTION

The invention will now be illustrated by tests carried out by the inventors.

10 Materials and Methods

Subjects and samples

The present study included a total of 64 patients with Meniere's disease and 29

15 patients with vestibular migraine. Patients were included in 6 centers: Granada University Hospital, Poniente de El Ejido Hospital, Almería, San Agustín de Linares Hospital, Getafe University Hospital, Santiago de Compostela Clinical Hospital, Salamanca University Hospital, Lugo Hospital, between November from 2014 and March 2016. Peripheral blood samples were obtained for cell separation

20 mononuclear and cytokine determination after in vitro testing. These samples were obtained in patients who had not had vertigo crisis at present, nor had they presented it at least 30 days before the date of obtaining the sample. Patients were diagnosed using the diagnostic scale of the American Academy of Otolaryngology-Head and Neck Surgery (AAO-HNS) (Committee on Hearing and

25 Equilibrium guidelines for the diagnosis and evaluation of therapy in Meniere’s disease. American Academy of Otolaryngology – Head and Neck Foundation, Inc. Otolaryngol Head Neck Surg. 1995. 113, 181-185).

A total of 54 samples of Spanish volunteers from the cities of Almería, Granada,

30 Salamanca, Jaén, Lugo and Santiago de Compostela were included as healthy controls. Quality controls were used on the samples of the individuals to exclude those that did not exceed the quality filters (elimination of duplicate samples, extreme values).

A basic neurotological exam including pure tone audiometry, nystagmus in

35 primary position, gaze evoked nystagmus, cephalic agitation nystagmus and standard caloric test was performed in all patients. The auditory stage for each patient

With definitive MS, it is defined as the average of the four-tone average of 0.5, 1, 2 and 3 kHz according to the AAO-HNS criteria: stage 1, ≤ 25 dB; stage 2, 26-40dB, stage 3, 41-70dB, and stage 4,> 70 dB. The diagnosis of migraine was made using the criteria of the International Headache Society and the diagnosis of vestibular migraine through

5 criteria of the Barany Society (2012).

The study was carried out in accordance with the principles of the Helsinki Declaration for research with human beings. The hospital research committee approved the research protocol. Peripheral blood mononuclear cells were isolated by

10 gradient of Ficoll. All patients and healthy individuals gave their informed consent for the study.

In vitro test of mononuclear cells and measurement of cytokines in supernatant using Multiplex plates.

15 Peripheral blood mononuclear cells were isolated by Ficoll density gradients and cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, pyruvate and essential amino acids, plated at 1x106 cells / ml in 12-well plates and incubated 16h at 37 ° C at 7% CO2. At the end of the incubation the cells are

20 centrifuged, the supernatant was stored and the RNA was extracted from the cells.

Multiplex analysis

The levels of the cytokines IL-1β, IL-6, and TNFα were quantified simultaneously by

25 an immunoassay with magnetic beads (EMD Millipore, Billerica, MA, USA) using a Luminex 2000 reader (Luminex Corp., Austin, TX, USA) and analyzed with Luminex x PONENT 3.1 software (Luminex Corp., Austin, TX, USES).

ELISAS

The levels of IL-1β and Il-6 were determined in supernatant by a sandwich ELISA (R&D Systems, Minneapolis, MN, USA) as described in the manufacturer's protocol. All samples were made in duplicate to ensure reproducibility. .

35 The results obtained by Multiplex and ELISA were compared for validation.

Statistic analysis.

A descriptive statistical analysis was done using the SPSS v.22 program (SPSS Inc, Chicago, IL, USA). The variables were compared using the T-student test. 5 A cluster analysis was carried out using the “gplots” package with the Heatmap.2 function of the R studio program.

Results

10 Two subgroups of patients were observed according to baseline levels of proinflammatory cytokines. Forty-two patients (68%) had low cytokine levels (IL-1β = 1.55 ± 1.53; IL-6 = 4.62 ± 4.99 and TNFα = 7.53 ± 6.17) while 20 patients (32%) exhibited elevated baseline levels of cytokines (IL-1β = 28.86 ± 12.84; IL-6 = 153.26 ± 115.26 and TNFα = 47.22 ± 16.01).

15 Individuals with vestibular migraine ((IL-1β = 1.4 ± 2.2; IL-6 = 4.8 ± 5.6 and TNFα = 5.9 ± 3.9) had cytokine levels very similar to that of healthy controls ((IL-1β = 2.3 ± 1.92 ; IL-6 = 4.8 ± 6.1 and TNFα = 5.32 ± 6.35).

The diagnostic profitability of IL-1β was assessed to discriminate between MV and MS, obtaining

20 a sensitivity of 35%, a specificity of 89%, a positive predictive value of 88% and a negative predictive value of 65%.

Claims (8)

1. A method of obtaining useful data for the diagnosis and classification of a disease that occurs with episodic vestibular syndrome, which includes: a) detect the levels of IL1β, IL6 and TNFα proinflammatory cytokines in an isolated biological sample from an individual, and b) compare the levels of proinflammatory cytokines detected in the step (a), with baseline levels in a healthy individual. 2. A method for the diagnosis and classification of a disease that develops with episodic vestibular syndrome comprises steps (a) and (b) according to the preceding claim, and further comprises: c) include the individual in the group of individuals with MS of autoimmune origin. 3. The method according to any of claims 1-2, wherein the sample isolated in step (a) is peripheral blood. The method according to any of claims 1-3, wherein the sample isolated in step (a) are mononuclear cells obtained from peripheral blood. 5. The method according to any of claims 1-4, wherein the cytokine determination of the invention is performed by techniques based on the antigen-antibody interaction. 6. The method according to any of claims 1-5, wherein the cytokine determination of the invention is performed by immunoassay, western blotting, immunoprecipitation, immunohistochemistry or immunofluorescence. The use of an immunosuppressant in the preparation of a medicament for the treatment of an individual from the group of individuals with MS of autoimmune origin identified by the diagnostic method according to any of claims 1-6. 8. A kit or device for the diagnosis and classification of a disease that is with episodic vestibular syndrome comprises antibodies capable of detecting and quantify the proinflammatory cytokines IL1β, IL6 and TNFα in an isolated biological sample, preferably peripheral blood, and even more preferably, peripheral blood mononuclear cells. 9. The use of a kit or device according to the previous reinvindication, for diagnosis and classification of a disease with episodic vestibular syndrome. 10. The use of a kit or device according to reinvindication 9, where the disease that occurs with episodic vestibular syndrome is MS. The use of a kit or device according to any of claims 9-10, wherein the disease that occurs with episodic vestibular syndrome is autoimmune MS. Figure 1 Figure 2 Figure 3 Figure 4 / LVWDGRGH6HFXHQFLDV <110> ANDALUS HEALTH SERVICE <120> Proinflammatory cytokines as a diagnostic marker in the episodic vestibular syndrome. <130> FIBAO-16004 <160> 14 <170> PatentIn version 3.5 <210> 1 <211> 1692 <212> DNA <213> Homo sapiens <400> 1 gtgtctgaag cagccatggc agaagtacct gagctcgcca gtgaaatgat ggcttattac 60 agtggcaatg aggatgactt gttctttgaa gctgatggcc ctaaacagat gaagtgctcc 120 ttccaggacc tggacctctg ccctctggat ggcggcatcc agctacgaat ctccgaccac 180 cactacagca agggcttcag gcaggccgcg tcagttgttg tggccatgga caagctgagg 240 aagatgctgg ttccctgccc acagaccttc caggagaatg acctgagcac cttctttccc 300 ttcatctttg aagaaggtag ttagccaaga gcaggcagta gatctccact tgtgtcctct 360 tggaagtcat caagccccag ccaactcaat tcccccagag ccaaagccct ttaaaggtag 420 aaggcccagc ggggagacaa aacaaagaag gctggaaacc aaagcaatca tctctttagt 480 ggaaactatt cttaaagaag atcttgatgg ctactgacat ttgcaactcc ctcactcttt 540 ctcaggggcc tttcacttac attgtcacca gagaacctat cttcttcgac acatgggata 600 acgaggctta tgtgcacgat gcacctgtac gatcactgaa ctgcacgctc cgggactcac 660 agcaaaaaag cttggtgatg tctggtccat atgaactgaa agctctccac ctccagggac 720 aggatatgga gcaacaagtg gtgttctcca tgtcctttgt acaaggagaa gaaagtaatg 780 acaaaatacc tgtggccttg ggcctcaagg aaaagaatct gtacctgtcc tgcgtgttga 840 aagatgataa gcccactcta cagctggaga gtgtagatcc caaaaattac ccaaagaaga 900 agatggaaaa gcgatttgtc ttcaacaaga tagaaatcaa taacaagctg gaatttgagt 960 ctgcccagtt ccccaactgg tacatcagca cctctcaagc agaaaacatg cccgtcttcc 1020 tgggagggac caaaggcggc caggatataa ctgacttcac catgcaattt gtgtcttcct 1080 aaagagagct gtacccagag agtcctgtgc tgaatgtgga ctcaatccct agggctggca 1140 gaaagggaac agaaaggttt ttgagtacgg ctatagcctg gactttcctg ttgtctacac 1200 caatgcccaa ctgcctgcct tagggtagtg ctaagaggat ctcctgtcca tcagccagga 1260 cagtcagctc tctcctttca gggccaatcc ccagcccttt tgttgagcca ggcctctctc 1320 acctctccta ctcacttaaa gcccgcctga cagaaaccac ggccacattt ggttctaaga 1380 aaccctctgt cattcgctcc cacattctga tgagcaaccg cttccctatt tatttattta 1440 tttgtttgtt tgttttattc attggtctaa tttattcaaa gggggcaaga agtagcagtg 1500

tctgtaaaag agcctagttt ttaatagcta tggaatcaat tcaatttgga ctggtgtgct

1560

ctctttaaat caagtccttt aattaagact gaaaatatat aagctcagat tatttaaatg

1620

ggaatattta taaatgagca aatatcatac tgttcaatgg ttctgaaata aacttcactg

1680

aagaaaaaaa aa

1692

<210> <211> <212> <213>

2 191 PRT Homo sapiens

<400>

2

Met Wing Thr Asp Ile Cys Asn Ser Leu Thr Leu Ser Gln Gly Pro Phe 1 5 10 15

Thr Tyr Ile Val Thr Arg Glu Pro Ile Phe Phe Asp Thr Trp Asp Asn 20 25 30

Glu Ala Tyr Val His Asp Ala Pro Val Arg Ser Leu Asn Cys Thr Leu 35 40 45

Arg Asp Ser Gln Gln Lys Ser Leu Val Met Ser Gly Pro Tyr Glu Leu 50 55 60

Lys Ala Leu His Leu Gln Gly Gln Asp Met Glu Gln Gln Val Val Phe 65 70 75 80

Be Met Be Phe Val Gln Gly Glu Glu Ser Asn Asp Lys Ile Pro Val 85 90 95

Wing Leu Gly Leu Lys Glu Lys Asn Leu Tyr Leu Ser Cys Val Leu Lys 100 105 110

Asp Asp Lys Pro Thr Leu Gln Leu Glu Ser Val Asp Pro Lys Asn Tyr 115 120 125

Pro Lys Lys Lys Met Glu Lys Arg Phe Val Phe Asn Lys Ile Glu Ile 130 135 140

Asn Asn Lys Leu Glu Phe Glu Ser Ala Gln Phe Pro Asn Trp Tyr Ile 145 150 155 160

Ser Thr Ser Gln Ala Glu Asn Met Pro Val Phe Leu Gly Gly Thr Lys 165 170 175

Gly Gly Gln Asp Ile Thr Asp Phe Thr Met Gln Phe Val Ser Ser 180 185 190

<210> <211> <212> <213>

3 1498 DNA Homo sapiens

<400> 3 accaaacctc ttcgaggcac aaggcacaac aggctgctct gggattctct tcagccaatc 60 ttcattgctc aagtgtctga agcagccatg gcagaagtac ctgagctcgc cagtgaaatg 120 atggcttatt acagtggcaa tgaggatgac ttgttctttg aagctgatgg ccctaaacag 180 atgaagtgct ccttccagga cctggacctc tgccctctgg atggcggcat ccagctacga 240 atctccgacc accactacag caagggcttc aggcaggccg cgtcagttgt tgtggccatg 300 gacaagctga ggaagatgct ggttccctgc ccacagacct tccaggagaa tgacctgagc 360 accttctttc ccttcatctt tgaagaagaa cctatcttct tcgacacatg ggataacgag 420 gcttatgtgc acgatgcacc tgtacgatca ctgaactgca cgctccggga ctcacagcaa 480 aaaagcttgg tgatgtctgg tccatatgaa ctgaaagctc tccacctcca gggacaggat 540 atggagcaac aagtggtgtt ctccatgtcc tttgtacaag gagaagaaag taatgacaaa 600 atacctgtgg ccttgggcct caaggaaaag aatctgtacc tgtcctgcgt gttgaaagat 660 gataagccca ctctacagct ggagagtgta gatcccaaaa attacccaaa gaagaagatg 720 gaaaagcgat ttgtcttcaa caagatagaa atcaataaca agctggaatt tgagtctgcc 780 cagttcccca actggtacat cagcacctct caagcagaaa acatgcccgt cttcctggga 840 gggaccaaag gcggccagga tataactgac ttcaccatgc aatttgtgtc ttcctaaaga 900 gagctgtacc cagagagtcc tgtgctgaat gtggactcaa tccctagggc tggcagaaag 960 ggaacagaaa ggtttttgag tacggctata gcctggactt tcctgttgtc tacaccaatg 1020 cccaactgcc tgccttaggg tagtgctaag aggatctcct gtccatcagc caggacagtc 1080 agctctctcc tttcagggcc aatccccagc ccttttgttg agccaggcct ctctcacctc 1140 tcctactcac ttaaagcccg cctgacagaa accacggcca catttggttc taagaaaccc 1200 tctgtcattc gctcccacat tctgatgagc aaccgcttcc ctatttattt atttatttgt 1260 ttgtttgttt tattcattgg tctaatttat tcaaaggggg caagaagtag cagtgtctgt 1320 aaaagagcct agtttttaat agctatggaa tcaattcaat ttggactggt gtgctctctt 1380 taaatcaagt cctttaatta agactgaaaa tatataagct cagattattt aaatgggaat 1440 atttataaat gagcaaatat catactgttc aatggttctg aaataaactt cactgaag 1498 <210> 4 <211> 269 <212> PRT <213> Homo sapiens <400> 4 Met Wing Glu Val Pro Glu Leu Wing Ser Glu Met Met Wing Tyr Tyr Ser 1 5 10 15 Gly Asn Glu Asp Asp Leu Phe Phe Glu Ala Asp Gly Pro Lys Gln Met 20 25 30 Lys Cys Ser Phe Gln Asp Leu Asp Leu Cys Pro Leu Asp Gly Gly Ile 35 40 45

Gln Leu Arg Ile Ser Asp His His Tyr Ser Lys Gly Phe Arg Gln Ala 50 55 60

Wing Ser Val Val Val Wing Met Asp Lys Leu Arg Lys Met Leu Val Pro 65 70 75 80

Cys Pro Gln Thr Phe Gln Glu Asn Asp Leu Ser Thr Phe Phe Pro Phe 85 90 95

Ile Phe Glu Glu Glu Pro Ile Phe Phe Asp Thr Trp Asp Asn Glu Ala 100 105 110

Tyr Val His Asp Wing Pro Val Arg Ser Leu Asn Cys Thr Leu Arg Asp 115 120 125

Ser Gln Gln Lys Ser Leu Val Met Ser Gly Pro Tyr Glu Leu Lys Wing 130 135 140

Leu His Leu Gln Gly Gln Asp Met Glu Gln Gln Val Val Phe Ser Met 145 150 155 160

Ser Phe Val Gln Gly Glu Glu Ser Asn Asp Lys Ile Pro Val Ala Leu 165 170 175

Gly Leu Lys Glu Lys Asn Leu Tyr Leu Ser Cys Val Leu Lys Asp Asp 180 185 190

Lys Pro Thr Leu Gln Leu Glu Ser Val Asp Pro Lys Asn Tyr Pro Lys 195 200 205

Lys Lys Met Glu Lys Arg Phe Val Phe Asn Lys Ile Glu Ile Asn Asn 210 215 220

Lys Leu Glu Phe Glu Ser Ala Gln Phe Pro Asn Trp Tyr Ile Ser Thr 225 230 235 240

Ser Gln Ala Glu Asn Met Pro Val Phe Leu Gly Gly Thr Lys Gly Gly 245 250 255

Gln Asp Ile Thr Asp Phe Thr Met Gln Phe Val Ser Ser 260 265

<210> <211> <212> <213>

5 1969 DNA Homo sapiens

<400> 5 gtattgtgtc actcagttca agtacttgaa atttattgaa ttgtattttc taaaaaatag

60

atagttgagt aaaagcaagc tcacattaca tagacggatc acagtgcacg gctgcggagc

120

tgggagcagt ggcttcgttt catgcaggaa agagaacttg gttcaggagt gtctacgttg

180

cttaagacag gagagcacta aaaatgaaac catccagcca tcctccccca ttttcatttt 240 cacaccaaag aatcccaccg cggcagagga ccaccgtctc tgtttagaca atcggtgaag 300 aatggatgac ctcactttcc ccaacaggcg ggtcctgaaa tgttatgcac gaaacaaaac 360 ttgagtaaat gcccaacaga ggtcactgtt ttatcgatct tgaagagatc tcttcttagc 420 aaagcaaaga aaccgattgt gaaggtaaca ccatgtttgg taaataagtg ttttggtgtt 480 gtgcaagggt ctggtttcag cctgaagcca tctcagagct gtctgggtct ctggagactg 540 gagggacaac ctagtctaga gcccatttgc atgagaccaa ggatcctcct gcaagagaca 600 ccatcctgag ggaagagggc ttctgaacca gcttgaccca ataagaaatt cttgggtgcc 660 gacgcggaag cagattcaga gcctagagcc gtgcctgcgt ccgtagtttc cttctagctt 720 cttttgattt caaatcaaga cttacaggga gagggagcga taaacacaaa ctctgcaaga 780 tgccacaagg tcctcctttg acatccccaa caaagaggac tggagatgtc tgaggctcat 840 tctgccctcg agcccaccgg gaacgaaaga gaagctctat ctcccctcca ggagcccagc 900 tatgaactcc ttctccacaa gcgccttcgg tccagttgcc ttctccctgg ggctgctcct 960 ggtgttgcct gctgccttcc ctgccccagt acccccagga gaagattcca aagatgtagc 1020 cgccccacac agacagccac tcacctcttc agaacgaatt gacaaacaaa ttcggtacat 1080 cctcgacggc atctcagccc tgagaaagga gacatgtaac aagagtaaca tgtgtgaaag 1140 cagcaaagag gcactggcag aaaacaacct gaaccttcca aagatggctg aaaaagatgg 1200 atgcttccaa tctggattca atgaggagac ttgcctggtg aaaatcatca ctggtctttt 1260 ggagtttgag gtatacctag agtacctcca gaacagattt gagagtagtg aggaacaagc 1320 cagagctgtg cagatgagta caaaagtcct gatccagttc ctgcagaaaa aggcaaagaa 1380 tctagatgca ataaccaccc ctgacccaac cacaaatgcc agcctgctga cgaagctgca 1440 ggcacagaac cagtggctgc aggacatgac aactcatctc attctgcgca gctttaagga 1500 gttcctgcag tccagcctga gggctcttcg gcaaatgtag catgggcacc tcagattgtt 1560 gttgttaatg ggcattcctt cttctggtca gaaacctgtc cactgggcac agaacttatg 1620 ttgttctcta tggagaacta aaagtatgag cgttaggaca ctattttaat tatttttaat 1680 ttattaatat ttaaatatgt gaagctgagt taatttatgt aagtcatatt tatattttta 1740 agaagtacca cttgaaacat tttatgtatt agttttgaaa taataatgga aagtggctat 1800 gcagtttgaa tatcctttgt ttcagagcca gatcatttct tggaaagtgt aggcttacct 1860 caaataaatg gctaacttat acatattttt aaagaaatat ttatattgta tttatataat 1920 gtataaatgg tttttatacc aataaatggc attttaaaaa attcagcaa 1969 <210> 6 <211> 212 <212> PRT <213> Homo sapiens <400> 6 Met Asn Ser Phe Ser Thr Ser Ala Phe Gly Pro Val Ala Phe Ser Leu 1 5 10 15

Gly Leu Leu Leu Val Leu Pro Wing Wing Phe Pro Wing Pro Val Pro Pro 20 25 30

Gly Glu Asp Ser Lys Asp Val Ala Ala Pro His Arg Gln Pro Leu Thr 35 40 45

Be Ser Glu Arg Ile Asp Lys Gln Ile Arg Tyr Ile Leu Asp Gly Ile 50 55 60

Ser Ala Leu Arg Lys Glu Thr Cys Asn Lys Ser Asn Met Cys Glu Ser 65 70 75 80

Ser Lys Glu Ala Leu Ala Glu Asn Asn Leu Asn Leu Pro Lys Met Ala 85 90 95

Glu Lys Asp Gly Cys Phe Gln Ser Gly Phe Asn Glu Glu Thr Cys Leu 100 105 110

Val Lys Ile Ile Thr Gly Leu Leu Glu Phe Glu Val Tyr Leu Glu Tyr 115 120 125

Leu Gln Asn Arg Phe Glu Ser Ser Glu Glu Gln Ala Arg Ala Val Gln 130 135 140

Met Ser Thr Lys Val Leu Ile Gln Phe Leu Gln Lys Lys Ala Lys Asn 145 150 155 160

Leu Asp Wing Ile Thr Thr Pro Asp Pro Thr Thr Asn Wing Ser Leu Leu 165 170 175

Thr Lys Leu Gln Wing Gln Asn Gln Trp Leu Gln Asp Met Thr Thr His 180 185 190

Leu Ile Leu Arg Ser Phe Lys Glu Phe Leu Gln Ser Ser Leu Arg Wing 195 200 205

Leu Arg Gln Met 210

<210> <211> <212> <213>

7 2555 DNA Homo sapiens

<400> 7 cccgctctgg ccccaccctc accctccaac aaagatttat caaatgtggg attttcccat

60

gagtctcaat attagagtct caacccccaa taaatatagg actggagatg tctgaggctc

120

attctgccct cgagcccacc gggaacgaaa gagaagctct atctcccctc caggagccca

180

gctatgaact ccttctccac aagtaagtgc aggaaatcct tagccctgga actgccagcg

240

gcggtcgagc cctgtgtgag ggaggggtgt gtggcccagg gagggctggc gggcggccag 300 cagcagaggc aggctcccag ctgtgctgtc agctcacccc tgcgctcgct cccctccggc 360 acaggcgcct tcggtccagt tgccttctcc ctggggctgc tcctggtgtt gcctgctgcc 420 ttccctgccc cagtaccccc aggagaagat tccaaagatg tagccgcccc acacagacag 480 ccactcacct cttcagaacg aattgacaaa caaattcggt acatcctcga cggcatctca 540 gccctgagaa aggagacatg taacaagagt aacatgtgtg aaagcagcaa agaggcactg 600 gcagaaaaca acctgaacct tccaaagatg gctgaaaaag atggatgctt ccaatctgga 660 ttcaatgagg agacttgcct ggtgaaaatc atcactggtc ttttggagtt tgaggtatac 720 ctagagtacc tccagaacag atttgagagt agtgaggaac aagccagagc tgtgcagatg 780 agtacaaaag tcctgatcca gttcctgcag aaaaaggtgg gtgtgtcctc attccctcaa 840 cttggtgtgg gggaagacag gctcaaagac agtgtcctgg acaactcagg gatgcaatgc 900 cacttccaaa agagaaggct acacgtaaac aaaagagtct gagaaatagt ttctgattgt 960 tattgttaaa tctttttttg tttgtttggt tggttggctc tcttctgcaa aggacatcaa 1020 taactgtatt ttaaactata tattaactga ggtggatttt aacatcaatt tttaatagtg 1080 caagagattt aaaaccaaag gcgggggggc gggcagaaaa aagtgcatcc aactccagcc 1140 agtgatccac agaaacaaag accaaggagc acaaaatgat tttaagattt tagtcattgc 1200 caagtgacat tcttctcact gtggttgttt caattctttt tcctaccttt taccagagag 1260 ttagttcaga gaaatggtca gagactcaag ggtggaaaga ggtaccaaag gctttggcca 1320 ccagtagctg gctattcaga cagcagggag tagacttgct ggctagcatg tggaggagcc 1380 aaagctcaat aagaaggggc ctagaatgaa acccttggtg ctgatcctgc ctctgccatt 1440 tctacttaag ccagggtttc tcatatgtta acatgcatgg gaattccctg ggcatcttct 1500 tgtggtgtgg agtctgactt agcaagcctc gggtgggttt gagggtcaaa tttctaccag 1560 gcttatatcc ctggtgatgc tgcagaattc caggaccaca cttggaggtt taaggccttc 1620 cacaagttac ttatcccata tggtgggtct atggaaaggt gtttcccagt cctctttaca 1680 ccaccggatc agtggtcttt caacagatcc taaagggatg gtgagaggga aactggagaa 1740 aagtatcaga tttagaggcc actgaagaac ccatattaaa atgcctttaa gtatgggctc 1800 ttcattcata tactaaatat gaactatgtg ccaggcatta tttcatatga cagaatacaa 1860 acaaataaga tagtgatgct ggtcaggctt ggtggctcat gcctgtattc cctaaacttt 1920 gggagcctaa ggtgagaact ccttgaactc ctaaggccag gagttcaaga ccagcctgga 1980 taacatagca agaccccatc tctacaaaaa accaaaacca aacaaacaaa aatgatagtg 2040 gtgcttccct caggatgctt gtggtctaat gggagacaga acagcaaagg gatgattaga 2100 agttggttgc tgtgagccag gcacagtgct gatataatcc cagcgctatg ggaggctgag 2160 gtgggtggat catttgaggc caggagttta agaccagcct ggtcaacatg gtaaaacccc 2220 atctctactt aaaaatacaa aaaagttagc caggcatggt ggcatacacc tgtaacccag 2280

ctactcagga ggctgaggca catgaatcac ttgaacccag gaggcagagg ttgctgtgca

2340

ccactgcact ccagcctggg tgacagaacg agaccttgac tcaaaaaaaa aaaaaagaag

2400

tttgttgcta tggaagggtc ctactcagag caggcacccc agttaatctc attcacccca

2460

catttcacat ttgaacatca tcccatagcc cagagcatcc ctccactgca aaggatttat

2520

tcaacattta aacaatcctt tttactttca ttttc

2555

<210> <211> <212> <213>

8 252 PRT Homo sapiens

<400>

8

Met Asn Ser Phe Ser Thr Ser Lys Cys Arg Lys Ser Leu Ala Leu Glu 1 5 10 15

Leu Pro Wing Wing Val Glu Pro Cys Val Arg Glu Gly Cys Val Wing Gln 20 25 30

Gly Gly Leu Wing Gly Gly Gln Gln Gln Arg Gln Wing Pro Ser Cys Wing 35 40 45

Val Ser Ser Pro Leu Arg Ser Leu Pro Ser Gly Thr Gly Ala Phe Gly 50 55 60

Pro Val Wing Phe Ser Leu Gly Leu Leu Leu Val Leu Pro Wing Wing Phe 65 70 75 80

Pro Ala Pro Val Pro Pro Gly Glu Asp Ser Lys Asp Val Ala Ala Pro 85 90 95

His Arg Gln Pro Leu Thr Ser Ser Glu Arg Ile Asp Lys Gln Ile Arg 100 105 110

Tyr Ile Leu Asp Gly Ile Ser Ala Leu Arg Lys Glu Thr Cys Asn Lys 115 120 125

Be Asn Met Cys Glu Be Be Lys Glu Ala Leu Ala Glu Asn Asn Leu 130 135 140

Asn Leu Pro Lys Met Wing Glu Lys Asp Gly Cys Phe Gln Ser Gly Phe 145 150 155 160

Asn Glu Glu Thr Cys Leu Val Lys Ile Ile Thr Gly Leu Leu Glu Phe 165 170 175

Glu Val Tyr Leu Glu Tyr Leu Gln Asn Arg Phe Glu Ser Ser Glu Glu 180 185 190

Gln Wing Arg Wing Val Gln Met Ser Thr Lys Val Leu Ile Gln Phe Leu 195 200 205

Gln Lys Lys Val Gly Val Ser Ser Phe Pro Gln Leu Gly Val Gly Glu 210 215 220 Asp Arg Leu Lys Asp Ser Val Leu Asp Asn Ser Gly Met Gln Cys His 225 230 235 240 Phe Gln Lys Arg Arg Leu His Val Asn Lys Arg Val 245 250 <210> 9 <211> 1201 <212> DNA <213> Homo sapiens <400> 9 aatattagag tctcaacccc caataaatat aggactggag atgtctgagg ctcattctgc 60 cctcgagccc accgggaacg aaagagaagc tctatctccc ctccaggagc ccagctatga 120 actccttctc cacaagcgcc ttcggtccag ttgccttctc cctggggctg ctcctggtgt 180 tgcctgctgc cttccctgcc ccagtacccc caggagaaga ttccaaagat gtagccgccc 240 cacacagaca gccactcacc tcttcagaac gaattgacaa acaaattcgg tacatcctcg 300 acggcatctc agccctgaga aaggagacat gtaacaagag taacatgtgt gaaagcagca 360 aagaggcact ggcagaaaac aacctgaacc ttccaaagat ggctgaaaaa gatggatgct 420 tccaatctgg attcaatgag gagacttgcc tggtgaaaat catcactggt cttttggagt 480 ttgaggtata cctagagtac ctccagaaca gatttgagag tagtgaggaa caagccagag 540 ctgtgcagat gagtacaaaa gtcctgatcc agttcctgca gaaaaaggca aagaatctag 600 atgcaataac cacccctgac ccaaccacaa atgccagcct gctgacgaag ctgcaggcac 660 agaaccagtg gctgcaggac atgacaactc atctcattct gcgcagcttt aaggagttcc 720 tgcagtccag cctgagggct cttcggcaaa tgtagcatgg gcacctcaga ttgttgttgt 780 taatgggcat tccttcttct ggtcagaaac ctgtccactg ggcacagaac ttatgttgtt 840 ctctatggag aactaaaagt atgagcgtta ggacactatt ttaattattt ttaatttatt 900 aatatttaaa tatgtgaagc tgagttaatt tatgtaagtc atatttatat ttttaagaag 960 taccacttga aacattttat gtattagttt tgaaataata atggaaagtg gctatgcagt 1020 ttgaatatcc tttgtttcag agccagatca tttcttggaa agtgtaggct tacctcaaat 1080 aaatggctaa cttatacata tttttaaaga aatatttata ttgtatttat ataatgtata 1140 aatggttttt ataccaataa atggcatttt aaaaaattca gcaaaaaaaa aaaaaaaaaa 1200 to 1201 <210> 10 <211> 212 <212> PRT <213> Homo sapiens <400> 10 Met Asn Ser Phe Ser Thr Ser Ala Phe Gly Pro Val Ala Phe Ser Leu 1 5 10 15

Gly Leu Leu Leu Val Leu Pro Wing Wing Phe Pro Wing Pro Val Pro Pro 20 25 30

Gly Glu Asp Ser Lys Asp Val Ala Ala Pro His Arg Gln Pro Leu Thr 35 40 45

Be Ser Glu Arg Ile Asp Lys Gln Ile Arg Tyr Ile Leu Asp Gly Ile 50 55 60

Ser Ala Leu Arg Lys Glu Thr Cys Asn Lys Ser Asn Met Cys Glu Ser 65 70 75 80

Ser Lys Glu Ala Leu Ala Glu Asn Asn Leu Asn Leu Pro Lys Met Ala 85 90 95

Glu Lys Asp Gly Cys Phe Gln Ser Gly Phe Asn Glu Glu Thr Cys Leu 100 105 110

Val Lys Ile Ile Thr Gly Leu Leu Glu Phe Glu Val Tyr Leu Glu Tyr 115 120 125

Leu Gln Asn Arg Phe Glu Ser Ser Glu Glu Gln Ala Arg Ala Val Gln 130 135 140

Met Ser Thr Lys Val Leu Ile Gln Phe Leu Gln Lys Lys Ala Lys Asn 145 150 155 160

Leu Asp Wing Ile Thr Thr Pro Asp Pro Thr Thr Asn Wing Ser Leu Leu 165 170 175

Thr Lys Leu Gln Wing Gln Asn Gln Trp Leu Gln Asp Met Thr Thr His 180 185 190

Leu Ile Leu Arg Ser Phe Lys Glu Phe Leu Gln Ser Ser Leu Arg Wing 195 200 205

Leu Arg Gln Met 210

<210> <211> <212> <213>

11 1006 DNA Homo sapiens

<400> 11 gtctcaatat tagagtctca acccccaata aatataggac tggagatgtc tgaggctcat

60

tctgccctcg agcccaccgg gaacgaaaga gaagctctat ctcccctcca ggagcccagc

120

tatgaactcc ttctccacaa acatgtaaca agagtaacat gtgtgaaagc agcaaagagg

180

cactggcaga aaacaacctg aaccttccaa agatggctga aaaagatgga tgcttccaat

240

ctggattcaa tgaggagact tgcctggtga aaatcatcac tggtcttttg gagtttgagg 300 tatacctaga gtacctccag aacagatttg agagtagtga ggaacaagcc agagctgtgc 360 agatgagtac aaaagtcctg atccagttcc tgcagaaaaa ggcaaagaat ctagatgcaa 420 taaccacccc tgacccaacc acaaatgcca gcctgctgac gaagctgcag gcacagaacc 480 agtggctgca ggacatgaca actcatctca ttctgcgcag ctttaaggag ttcctgcagt 540 ccagcctgag ggctcttcgg caaatgtagc atgggcacct cagattgttg ttgttaatgg 600 gcattccttc ttctggtcag aaacctgtcc actgggcaca gaacttatgt tgttctctat 660 ggagaactaa aagtatgagc gttaggacac tattttaatt atttttaatt tattaatatt 720 taaatatgtg aagctgagtt aatttatgta agtcatattt atatttttaa gaagtaccac 780 ttgaaacatt ttatgtatta gttttgaaat aataatggaa agtggctatg cagtttgaat 840 atcctttgtt tcagagccag atcatttctt ggaaagtgta ggcttacctc aaataaatgg 900 ctaacttata catattttta aagaaatatt tatattgtat ttatataatg tataaatggt 960 ttttatacca ataaatggca ttttaaaaaa ttcagcaaaa aaaaaa 1006 <210> 12 <211> 136 <212> PRT <213> Homo sapiens <400> 12 Met Cys Glu Ser Ser Lys Glu Ala Leu Ala Glu Asn Asn Leu Asn Leu 1 5 10 15 Pro Lys Met Wing Glu Lys Asp Gly Cys Phe Gln Ser Gly Phe Asn Glu 20 25 30 Glu Thr Cys Leu Val Lys Ile Ile Thr Gly Leu Leu Glu Phe Glu Val 35 40 45 Tyr Leu Glu Tyr Leu Gln Asn Arg Phe Glu Ser Ser Glu Glu Gln Ala 50 55 60 Arg Wing Val Gln Met Ser Thr Lys Val Leu Ile Gln Phe Leu Gln Lys 65 70 75 80 Lys Wing Lys Asn Leu Asp Wing Ile Thr Thr Pro Asp Pro Thr Thr Asn 85 90 95 Ser Ser Leu Leu Thr Lys Leu Gln Gln Wing Asn Gln Trp Leu Gln Asp 100 105 110 Met Thr Thr His Leu Ile Leu Arg Ser Phe Lys Glu Phe Leu Gln Ser 115 120 125 Ser Leu Arg Ala Leu Arg Gln Met 130 135 <210> 13 <211> 1686 <212> DNA <213> Homo sapiens <400> 13 cagacgctcc ctcagcaagg acagcagagg accagctaag agggagagaa gcaactacag 60 accccccctg aaaacaaccc tcagacgcca catcccctga caagctgcca ggcaggttct 120 cttcctctca catactgacc cacggctcca ccctctctcc cctggaaagg acaccatgag 180 cactgaaagc atgatccggg acgtggagct ggccgaggag gcgctcccca agaagacagg 240 ggggccccag ggctccaggc ggtgcttgtt cctcagcctc ttctccttcc tgatcgtggc 300 aggcgccacc acgctcttct gcctgctgca ctttggagtg atcggccccc agagggaaga 360 gttccccagg gacctctctc taatcagccc tctggcccag gcagtcagat catcttctcg 420 aaccccgagt gacaagcctg tagcccatgt tgtagcaaac cctcaagctg aggggcagct 480 ccagtggctg aaccgccggg ccaatgccct cctggccaat ggcgtggagc tgagagataa 540 ccagctggtg gtgccatcag agggcctgta cctcatctac tcccaggtcc tcttcaaggg 600 ccaaggctgc ccctccaccc atgtgctcct cacccacacc atcagccgca tcgccgtctc 660 ctaccagacc aaggtcaacc tcctctctgc catcaagagc ccctgccaga gggagacccc 720 agagggggct gaggccaagc cctggtatga gcccatctat ctgggagggg tcttccagct 780 ggagaagggt gaccgactca gcgctgagat caatcggccc gactatctcg actttgccga 840 gtctgggcag gtctactttg ggatcattgc cctgtgagga ggacgaacat ccaaccttcc 900 caaacgcctc ccctgcccca atccctttat taccccctcc ttcagacacc ctcaacctct 960 tctggctcaa aaagagaatt gggggcttag ggtcggaacc caagcttaga actttaagca 1020 acaagaccac cacttcgaaa cctgggattc aggaatgtgt ggcctgcaca gtgaagtgct 1080 ggcaaccact aagaattcaa actggggcct ccagaactca ctggggccta cagctttgat 1140 ccctgacatc tggaatctgg agaccaggga gcctttggtt ctggccagaa tgctgcagga 1200 cttgagaaga cctcacctag aaattgacac aagtggacct taggccttcc tctctccaga 1260 tgtttccaga cttccttgag acacggagcc cagccctccc catggagcca gctccctcta 1320 tttatgtttg cacttgtgat tatttattat ttatttatta tttatttatt tacagatgaa 1380 tgtatttatt tgggagaccg gggtatcctg ggggacccaa tgtaggagct gccttggctc 1440 agacatgttt tccgtgaaaa cggagctgaa caataggctg ttcccatgta gccccctggc 1500 ctctgtgcct tcttttgatt atgtttttta aaatatttat ctgattaagt tgtctaaaca 1560 atgctgattt ggtgaccaac tgtcactcat tgctgagcct ctgctcccca ggggagttgt 1620 gtctgtaatc gccctactat tcagtggcga gaaataaagt ttgcttagaa aagaaaaaaa 1680 aaaaaa 1686 <210> 14 <211> 233 <212> PRT <213> Homo sapiens <400> 14 Met Ser Thr Glu Ser Met Ile Arg Asp Val Glu Leu Ala Glu Glu Ala 1 5 10 15 Leu Pro Lys Lys Thr Gly Gly Pro Gln Gly Ser Arg Arg Cys Leu Phe 20 25 30 Leu Ser Leu Phe Ser Phe Leu Ile Val Ala Gly Ala Thr Thr Leu Phe 35 40 45 Cys Leu Leu His Phe Gly Val Ile Gly Pro Gln Arg Glu Glu Phe Pro 50 55 60 Arg Asp Leu Ser Leu Ile Ser Pro Leu Gln Wing Val Val Arg Ser Ser 65 70 75 80 Ser Arg Thr Pro Ser Asp Lys Pro Val Ala His Val Val Ala Asn Pro 85 90 95 Gln Wing Glu Gly Gln Leu Gln Trp Leu Asn Arg Arg Wing Asn Wing Leu 100 105 110 Leu Wing Asn Gly Val Glu Leu Arg Asp Asn Gln Leu Val Val Pro Ser 115 120 125 Glu Gly Leu Tyr Leu Ile Tyr Ser Gln Val Leu Phe Lys Gly Gln Gly 130 135 140 Cys Pro Ser Thr His Val Leu Leu Thr His Thr Ile Ser Arg Ile Ala 145 150 155 160 Val Ser Tyr Gln Thr Lys Val Asn Leu Leu Ser Ala Ile Lys Ser Pro 165 170 175 Cys Gln Arg Glu Thr Pro Glu Gly Ala Glu Ala Lys Pro Trp Tyr Glu 180 185 190 Pro Ile Tyr Leu Gly Gly Val Phe Gln Leu Glu Lys Gly Asp Arg Leu 195 200 205 Ser Ala Glu Ile Asn Arg Pro Asp Tyr Leu Asp Phe Ala Glu Ser Gly 210 215 220 Gln Val Tyr Phe Gly Ile Ile Ala Leu 225 230