Cytokine analysis may support new therapeutic strategies for immune-mediated hearing loss.

Cytokine analysis may support new therapeutic strategies for immune-mediated hearing loss. | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Cytokine analysis may support new therapeutic strategies for immune-mediated hearing loss. Jose Maria Verdaguer muñoz, Ana Sánchez-Martínez, Nuria Arnáiz-Canora, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4622777/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Purpose: Diagnosing immune-mediated hearing loss is challenging due to diverse clinical presentations and lack of specific markers. This study assesses cytokine levels in these patients to identify potential diagnostic markers and new treatments. Materials and Methods: A retrospective analysis of 70 early-stage immune-mediated hearing loss patients was conducted. Inclusion criteria included clinical presentation, immunological work-up alterations, corticosteroid response, and comparison with healthy controls. Serum levels of IFN gamma, IL-1Beta, IL-6, IL-10, IL-17a, and TNFalfa were measured using a Magnetic Luminex Assay kit. Results: The cohort, with an average age of 42.1 years, showed bilateral hearing loss in 30% of cases, predominantly as sudden sensorineural hearing loss (54.2%). Among cytokines analyzed, only IL-10 levels were elevated compared to controls. Discussion: Diagnosis relies on clinical evaluation due to limited diagnostic markers. Previous studies on cytokine involvement had conflicting results. Elevated IL-10 levels suggest a role in pathogenesis and treatment response, warranting further investigation. Conclusion: IL-10 therapy is proposed for corticosteroid non-responders, offering a promising research direction. Identifying diagnostic markers and understanding immune-related causes are crucial for improving outcomes in immune-mediated hearing loss, supporting ongoing research efforts. Biological sciences/Immunology/Applied immunology Biological sciences/Immunology/Cytokines Biological sciences/Immunology/Immunological disorders Biological sciences/Immunology/Neuroimmunology Health sciences/Diseases/Immunological disorders/Autoimmune diseases Immune-mediated hearing loss Cytokines Corticosteroid therapy IL-10 Autoimmune inner ear disease (AIED) Figures Figure 1 INTRODUCTION Immune-mediated hearing loss presents a rare and challenging diagnostic scenario, characterized by symptoms such as rapidly progressive hearing loss or sudden deafness, often followed by fluctuating hearing loss as the condition advances 1 . Consequently, long-term follow-up proves invaluable in establishing a suspected diagnosis. However, this clinical course closely resembles other inner ear disorders, including sudden deafness, Meniere's disease, and otosyphilis 2 – 4 , posing a considerable challenge for otolaryngologists. Currently, no specific marker has been identified for this condition, necessitating reliance on clinical experience and suspicion 5 . Nevertheless, efforts have been made to establish diagnostic criteria, with clinical presentation and response to treatments such as steroids and immunomodulators serving as key considerations 6 , 7 . Additionally, the presence of nuclear autoantibodies and elevated subpopulations of CD4CD45RO T helper lymphocytes have been noted to strengthen clinical suspicion 8 , 9 Immune-mediated hearing loss can occur as an isolated entity, representing the majority of cases, or in the context of a systemic autoimmune disease, with deafness being the manifestation that may appear as the first symptom. Primary cases pose greater difficulty in diagnosis as they may not have other clinical manifestations or a panel of autoantibodies as seen in systemic autoimmune diseases. Etiopathogenetically, the immune response within the inner ear would be responsible for the cellular and/or vascular damage that justifies the hearing loss. 10 , 11 . This response can be spontaneous or secondary to another event, such as a viral disease of the upper aerodigestive tract, which can also occur in systemic autoimmune diseases. Various cellular populations (B cells, T cells, etc.) and cytokines such as interferon gamma, TNF alpha, IL-6, etc. may mediate this immune response. 12 – 15 . The objective of this study is to determine the existence of various cytokines in the serum of patients affected by immune-mediated hearing loss, preserved in a bio bank, in order to establish their role in the etiopathogenesis of the immune response (and posterior use as diagnostic markers) as well as to search for possible new treatment alternatives. MATERIALS AND METHODS We conducted a retrospective study involving 70 patients diagnosed with immune-mediated hearing loss, alongside 54 healthy adult controls with no prior history of hearing impairment or autoimmune/autoinflammatory disorders. INCLUSION CRITERIA Immunomediated inner ear disease (IMIED) patients included in the present study fulfilled the inclusion criteria previously reported. 16 Patients had a diagnosis of IMIED if they had an episode of presence of sensorineural hearing loss manifested as a greater progress at any frequency and audiometric evidence of progression in at least one ear manifested as a threshold shift greater than 15 dB at any frequency or 10 dB at two or more frequencies on serial audiograms in > 3 days but less than 3 months apart. If the hearing loss developed in 1/160, altered T-cell immunophenotype, response to corticosteroids (oral or intratympanic) in each episode (except in cases with profound hearing loss), possible association with systemic autoimmune diseases. 17 , 18 . Healthy controls were adults without history of hearing loss and autoimmune/autoinflammatory disorders. Patients were treated with 60mg of oral prednisone or methylprednisolone for 3–4 weeks followed by a variable taper (2 months taper for rapidly progressive SNHL and 1–2 weeks for sudden SNHL). The pure-tone average included frequencies: 250, 500, 1000, 2000, 4000 Hz to determine corticosteroid response. Criteria of a response to corticosteroid therapy included the following: 1. For patients with rapidly progressive sensorineural hearing loss (> 3 days to < 90 days), improvement in the pure-tone average of 5 dB and a 15 dB improvement in 1 frequency or 10 dB at 2 frequencies, for at least 4 weeks while receiving the therapy. 19 Weekly audiograms were obtained during a 2-month period. 2. For patients with sudden sensorineural hearing loss (< 3 days): 3 improvement in the final pure-tone average of more than 10dB. Serial audiograms were obtained for 1 year. Samples and pertinent patient data utilized in this study were sourced from our Center's Biobank, adhering to standard operating protocols and with requisite approval from both Ethics and Scientific Committees. The samples from these patients were collected at the time of diagnosis after obtaining informed consent. Frozen serum samples were thawed prior to analysis, and none of them underwent more than two freeze-thaw cycles before analysis, following the instructions provided in the kit. The cytokines levels in serum were assessed using a Magnetic Luminex Assay kit (ME-HCYT-60K-06, Merck Millipore). A panel of six cytokines (IFN- γ, IL-1 β, IL-6, IL-10, IL-17A y TNF- α) was selected for analysis and measured in a Luminex 200 analyzer instrument according to the manufacturer’s protocol. Luminex data were analyzed using Xponent software. All procedures were conducted in compliance with the applicable regulations and guidelines of the Drug Research Ethics Committee at Puerta de Hierro Majadahonda University Hospital. The study received approval under protocol number 07/2024. All procedures were compliant with the ethical principles for medical research involving human subjects as outlined in the WMA Declaration of Helsinki. RESULTS Clinical Characteristics (Table 1 ): Table 1 Demographics and Clinical presentation Patients Controls SEX Female 35 29 Male 35 25 AGE Average age 42.1 41.6 CLINIC Sudden sensorineural hearing loss 38 (54.28%) Progressive hearing loss 25 (35.72%) Fluctuating hearing loss 7 (10%) This study encompassed a cohort of 70 patients diagnosed with IMIED. Among them, 35 (50%) were male, and 35 (50%) were female, with an average age of 42.1 years. The control group consisted of 54 specimens, comprising 29 (54%) females and 25 (46%) males, with an average age of 41.6 years. Bilateral hearing loss was observed in 21 patients (30%). Sudden sensorineural hearing loss was the most frequent clinical form of presentation (38 cases, 54.2%). IL-10 cytokine levels were found to be elevated in patients diagnosed with IMIED compared to the control group, although the results are not statistically significant. Conversely, the levels of other cytokines were higher in control patients than in those with IMIED with no statistically significant differences except for IL-6 (p = 0.025). (Table 2 and Fig. 1 ) Table 2 Cytokines results Cytokine Cytokine type p50 control p50 patients p75 control p75 patients Statistical test p-valor INF-γ Pro-inflammatory 5.9 4.3 30 9.6 Mann-Whitney U > 0.05 IL-1β Pro-inflammatory 0 1 13 3.4 Mann-Whitney U > 0.05 IL-6 Pro-inflammatory 1.9 0.97 4.8 3.2 Mann-Whitney U 0.05 IL-17 Pro-inflammatory 0 0 2.7 1.7 Mann-Whitney U > 0.05 TNF-α Pro-inflammatory 26 23 43 38 Mann-Whitney U > 0.05 DISCUSSION The diagnosis of idiopathic sudden sensorineural hearing loss (ISSNHL) still heavily relies on clinical evaluation in many cases, with only limited markers available to assist and confirm the diagnosis. The immunopathological mechanisms underlying this condition remain elusive. Due to the impracticality of biopsying the inner ear, which would result in its destruction, alternative investigative methods have been explored, such as MRI studies utilizing intratympanic gadolinium 20 . However, the ultimate pathophysiological or immunopathological mechanism has yet to be fully elucidated. In recent years, several articles have proposed the correlation and potential involvement of certain interleukins as immunopathological mechanisms in this condition 21 . However, currently, there are no cytokines identified to definitively support the diagnosis of this disease. The findings across published studies remain controversial. (Table 3 ) Table 3 Previous research on the correlation and potential involvement of interleukins in immune mediated inner ear disease Study Molecules Results Amor Dorado JC et al HLA DRA1∗04 Marker of autoimmune middle ear disease, worse prognosis associated. Psillas G. et al HLA DRA1∗04 favorable response to treatment (non statistically significant) SW Gorthey et al IL-6 Elevated IL6 levels are associated with a worse response to treatment. Moleon MDC et al IL-1β, CCL3, CCL4, CXCL1, CCL22, CCL8 Cytokine levels do not allow distinguishing between late-onset or early-onset Meniere's disease. There have been studies examining the association between HLA and IMIED, although they are limited in number 22 . Certain HLA types, such as HLA DRB104, have been identified as potential markers for autoimmune inner ear diseases 23 . HLA DRB104 has demonstrated prognostic significance in patients experiencing sudden sensorineural hearing loss, being linked to a poorer treatment outcome. However, in the study conducted by Psillas et al. 24 , was examined the correlation between HLA and response to corticosteroid treatment. In this instance, a favorable response to treatment was observed, although the outcome did not reach statistical significance. Gorthey et al. conducted an analysis investigating the correlation between IL-6 levels and the efficacy of corticosteroid treatment in individuals with autoimmune-related hearing loss. Their findings demonstrated that heightened IL-6 levels corresponded with a reduced response to treatment, offering significant insights for timely therapeutic interventions in such patients 12 . In our current study, IL-6 levels were observed to be higher in controls compared to patients. However, it is noteworthy that all patients exhibited a positive response to treatment, as the diagnosis was established in the early stages, and positive treatment response was a criterion for diagnosis. In 2020, Moleon et al. employed multiplex technology to quantify various cytokine levels in Meniere's disease, with the objective of delineating the cytokine profile based on the age of disease onset. The study did not yield statistically significant differences overall. However, when stratifying by gender, notable distinctions emerged: levels of ILB were found to be higher in men than in women during early onset, whereas levels of CXCL1 were higher in women compared to men. 4 In the present study, elevated levels of IL-10 were observed in individuals with autoimmune hearing loss. Moreover, given the favorable response to corticosteroid therapy exhibited by the study participants, this biomarker may serve as a predictive indicator for treatment response. IL-10 serves as a crucial anti-inflammatory cytokine with significant effects on antigen-presenting cells. It operates by impeding the synthesis of proinflammatory cytokines such as IL1-β, TNF-α, and IL-6, along with Th2 cell-derived cytokines like IL-4 and IL-5. Additionally, IL-10 exerts a direct regulatory influence on T cell differentiation by suppressing IL-2 and IFN-γ. 25 – 28 This observation offers a potential explanation for the elevated IL-10 levels in our patients alongside the comparatively lower levels of other analyzed cytokines in comparison to control subjects. Likewise, IL-10 also demonstrates higher expression in patients with idiopathic ulcerative colitis during remission. This upregulation is attributed to its anti-inflammatory properties within the intestine, where it acts to inhibit antigen presentation and the subsequent release of proinflammatory cytokines. 27 . In this context, the administration of recombinant IL-10 and gelatin microspheres incorporating IL-10 in ulcerative colitis has been suggested. 26 . Based on this finding, we propose a novel treatment strategy with IL-10 (transtympanic or systemically administrated) for corticosteroids non-responder IMIED patients. Since the inner ear lacks a lymphatic system the endolymphatic sac could potentially function as a mucosa-associated lymphoid tissue (MALT) organ within the inner ear 10 , transtympanic injection of IL-10 may represent a new therapeutic strategy in IMIED patients. Recently, Xinyuan Tan et al. 29 delve into the significance of the interplay between inflammatory and anti-inflammatory cytokines in the pathogenesis of sudden hearing loss, as well as our proposed theory on autoimmune hearing loss. This elucidates the favorable response to corticosteroid treatment, attributed to their anti-inflammatory properties. The findings from the experimental study conducted in mice by Zhou et al. provide support for the theory behind this novel treatment approach. Their results indicate that a deficiency in IL-10 worsens hearing loss, while the exogenous administration of IL-10 enhances hearing function. These outcomes underscore the pivotal role of IL-10 as a key regulator of inflammation in vivo. 30 Due to the extended duration of IMIED, which may lead to hearing fluctuations unresponsive to corticosteroids, there's a need to explore the role of cytokines in later stages. This prompts our ongoing research into the rationale behind IL-10 treatment for non-responders. CONCLUSIONS In recent years, studies have aimed to assess cytokine levels in patients with autoimmune hearing loss, with the goal of elucidating the etiopathogenesis of this condition. However, further research is required to fully understand the underlying mechanisms. The elevated levels of IL-10 observed in our patients may account for the absence of elevated levels of other analyzed cytokines, owing to its anti-inflammatory properties. Despite the limitations inherent in our retrospective study design, the substantial number of participants enables us to support the hypothesis regarding the potential role of IL-10 administration in patients who do not respond to corticosteroid treatment. Our findings suggest a promising avenue for further investigation into the efficacy of IL-10 therapy in this subset of patients. Declarations Author Contribution ASM: data collecting, writing and editing the first draftNAC: data collectingAR: biostatistical analysisJMVM: manuscript review, corresponding authorAJSL: data analysisSRG: data analysisJRGB: research coordinator, final approvalAcknowledgments “The authors wish to thank the donors, and the Hospital Universitario Puerta de Hierro Majadahonda (HUPHM)/Instituto de Investigación Sanitaria Puerta de Hierro-Segovia de Arana (IDIPHISA) Biobank (Carlos III Health Institute Biobanks and Biomodels Platform – PT23/00015) for the human specimens used in this study”. Acknowledgement The authors wish to thank the donors, and the Hospital Universitario Puerta de Hierro Majadahonda (HUPHM)/Instituto de Investigación Sanitaria Puerta de Hierro-Segovia de Arana (IDIPHISA) Biobank (Carlos III Health Institute Biobanks and Biomodels Platform – PT23/00015) for the human specimens used in this study. Data Availability The data that support the findings of this study are not openly available due to reasons of sensitivity and are available from the corresponding author upon reasonable request. References McCabe, B. F. Autoimmune sensorineural hearing loss. Ann. Otol. Rhinol. Laryngol. 88, 585–589 (1979). García-Berrocal, J. R. et al. Otosyphilis mimics immune disorders of the inner ear. Acta Otolaryngol. (Stockh.) 126, 679–684 (2006). Herrera, M., García Berrocal, J. R., García Arumí, A., Lavilla, M. J. & Plaza, G. Actualización del consenso sobre el diagnóstico y tratamiento de la sordera súbita idiopática. Acta Otorrinolaringológica Esp. 70, 290–300 (2019). Moleon, M.-D.-C. et al. Clinical and Cytokine Profile in Patients with Early and Late Onset Meniere Disease. J. Clin. Med. 10, 4052 (2021). Lobo, D., López, F. G., García-Berrocal, J. R. & Ramírez-Camacho, R. Diagnostic tests for immunomediated hearing loss: a systematic review. J. Laryngol. Otol. 122, 564–573 (2008). García-Berrocal, J. R. et al. Alternatives to systemic steroid therapy for refractory immune-mediated inner ear disease: A physiopathologic approach. Eur. Arch. Oto-Rhino-Laryngol. Off. J. Eur. Fed. Oto-Rhino-Laryngol. Soc. EUFOS Affil. Ger. Soc. Oto-Rhino-Laryngol. - Head Neck Surg. 263, 977–982 (2006). David R Lobo, J. R. G., a-Berrocal, R. R. & rez-Camacho. New prospects in the diagnosis and treatment of immune-mediated inner ear disease. World J. Methodol. 4, 91–98 (2014). Garcia-Berrocal, J. R. et al. Deficiency of Naive T Cells in Patients With Sudden Deafness. Arch. Otolaryngol. - Head Neck Surg. 123, 712–717 (1997). Lobo, D., García-Berrocal, J. R., Trinidad, A., Verdaguer, J. M. & Ramirez-Camacho, R. Review of the biologic agents used for immune-mediated inner ear disease. Acta Otorrinolaringol. Esp. (2012) doi: 10.1016/j.otorri.2012.04.008 . García Berrocal, J. R. & Ramírez-Camacho, R. Immune response and immunopathology of the inner ear: an update. J. Laryngol. Otol. 114, 101–107 (2000). Hashimoto, S., Billings, P., Harris, J. P., Firestein, G. S. & Keithley, E. M. Innate immunity contributes to cochlear adaptive immune responses. Audiol. Neurootol. 10, 35–43 (2005). Gorthey, S. W., Pathak, S. & Vambutas, A. The Correlation of Clinical Corticosteroid Responsiveness With Expression of IL-6 in Peripheral Blood Immune Cells (PBMC) in Patients With Autoimmune Inner Ear Disease (AIED). Otol. Neurotol. 42, 1422–1428 (2021). Lobo, D., Trinidad, A., García-Berrocal, J. R., Verdaguer, J. M. & Ramírez-Camacho, R. TNFalpha blockers do not improve the hearing recovery obtained with glucocorticoid therapy in an autoimmune experimental labyrinthitis. Eur. Arch. Oto-Rhino-Laryngol. Off. J. Eur. Fed. Oto-Rhino-Laryngol. Soc. EUFOS Affil. Ger. Soc. Oto-Rhino-Laryngol. - Head Neck Surg. 263, 622–626 (2006). Lorenz, R. R. et al. Interferon-g production to inner ear antigens by T cells from patients with autoimmune sensorineural hearing loss. J. Neuroimmunol. (2002). Vambutas, A. et al. Early efficacy trial of anakinra in corticosteroid-resistant autoimmune inner ear disease. J. Clin. Invest. 124, 4115–4122 (2014). Moscicki, R. A. et al. Serum antibody to inner ear proteins in patients with progressive hearing loss. Correlation with disease activity and response to corticosteroid treatment. JAMA 272, 611–616 (1994). Berrocal, J. R. G. & Ramírez-Camacho, R. Sudden sensorineural hearing loss: supporting the immunologic theory. Ann. Otol. Rhinol. Laryngol. 111, 989–997 (2002). García-Berrocal, J. R. et al. Sudden presentation of immune-mediated inner ear disease: characterization and acceptance of a cochleovestibular dysfunction. J. Laryngol. Otol. 117, 775–779 (2003). Niparko, J. K. et al. Serial Audiometry in a Clinical Trial of AIED Treatment. Otol. Neurotol. 26, 908–917 (2005). Lobo, D., Tuñón, M., Villarreal, I., Brea, B. & García-Berrocal, J. R. Intratympanic gadolinium magnetic resonance imaging supports the role of endolymphatic hydrops in the pathogenesis of immune-mediated inner-ear disease. J. Laryngol. Otol. 132, 554–559 (2018). Hajas, A. et al. Sensorineural Hearing Loss in Patients with Mixed Connective Tissue Disease: Immunological Markers and Cytokine Levels. J. Rheumatol. 36, 1930–1936 (2009). García-Berrocal, J. R. et al. Deficiency of naive T cells in patients with sudden deafness. Arch. Otolaryngol. Head Neck Surg. 123, 712–717 (1997). Amor-Dorado, J. C., Paco, L., Martin, J., Lopez-Nevot, M. A. & Gonzalez-Gay, M. A. Human leukocyte antigen-DQB1 and -DRB1 associations in patients with idiopathic sudden sensorineural hearing loss from a defined population of Northwest Spain. Acta Otolaryngol. (Stockh.) 125, 1277–1282 (2005). Psillas, G., Binos, P., Dimas, G. G., Daniilidis, M. & Constantinidis, J. Human Leukocyte Antigen (HLA) Influence on Prognosis of Autoimmune Hearing Loss. Audiol. Res. 11, 31–37 (2021). Ebert, E. C. IL-10 enhances IL-2-induced proliferation and cytotoxicity by human intestinal lymphocytes. Clin. Exp. Immunol. 119, 426–432 (2000). Fonseca-Camarillo, G., Furuzawa-Carballeda, J., Martínez-Benítez, B., Barreto-Zúñiga, R. & Yamamoto-Furusho, K. [Intelukin-10 expression with immunorreglatory function in the mucosa of patients with ulcerative colitis]. Rev. Gastroenterol. Mex. 76, 113–119 (2011). Li, M.-C. IL-10 and its related cytokines for treatment of inflammatory bowel disease. World J. Gastroenterol. 10, 620 (2004). Staples, K. J., Bergmann, M., Barnes, P. J. & Newton, R. Stimulus-Specific Inhibition of IL-5 by cAMP-Elevating Agents and IL-10 Reveals Differential Mechanisms of Action. Biochem. Biophys. Res. Commun. 273, 811–815 (2000). Tan, X. et al. Relevant Research of Inflammatory Cytokines Spectrum in Peripheral Blood of Sudden Hearing Loss. The Laryngoscope lary.31276 (2024) doi: 10.1002/lary.31276 . Zhou, B. et al. Experimental autoimmune hearing loss is exacerbated in IL-10-deficient mice and reversed by IL-10 gene transfer. Gene Ther. 19, 228–235 (2012). Additional Declarations No competing interests reported. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-4622777","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":326663380,"identity":"ba4539b6-c74b-426a-91aa-c590834e6667","order_by":0,"name":"Jose Maria Verdaguer 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20:50:23","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":564256,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4622777/v1/e914fadc-65aa-423b-b1fb-42eb338ea3a3.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Cytokine analysis may support new therapeutic strategies for immune-mediated hearing loss.","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eImmune-mediated hearing loss presents a rare and challenging diagnostic scenario, characterized by symptoms such as rapidly progressive hearing loss or sudden deafness, often followed by fluctuating hearing loss as the condition advances \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e. Consequently, long-term follow-up proves invaluable in establishing a suspected diagnosis. However, this clinical course closely resembles other inner ear disorders, including sudden deafness, Meniere's disease, and otosyphilis \u003csup\u003e\u003cspan additionalcitationids=\"CR3\" citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e, posing a considerable challenge for otolaryngologists. Currently, no specific marker has been identified for this condition, necessitating reliance on clinical experience and suspicion \u003csup\u003e\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Nevertheless, efforts have been made to establish diagnostic criteria, with clinical presentation and response to treatments such as steroids and immunomodulators serving as key considerations \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. Additionally, the presence of nuclear autoantibodies and elevated subpopulations of CD4CD45RO T helper lymphocytes have been noted to strengthen clinical suspicion \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eImmune-mediated hearing loss can occur as an isolated entity, representing the majority of cases, or in the context of a systemic autoimmune disease, with deafness being the manifestation that may appear as the first symptom. Primary cases pose greater difficulty in diagnosis as they may not have other clinical manifestations or a panel of autoantibodies as seen in systemic autoimmune diseases.\u003c/p\u003e \u003cp\u003eEtiopathogenetically, the immune response within the inner ear would be responsible for the cellular and/or vascular damage that justifies the hearing loss. \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. This response can be spontaneous or secondary to another event, such as a viral disease of the upper aerodigestive tract, which can also occur in systemic autoimmune diseases. Various cellular populations (B cells, T cells, etc.) and cytokines such as interferon gamma, TNF alpha, IL-6, etc. may mediate this immune response.\u003csup\u003e\u003cspan additionalcitationids=\"CR13 CR14\" citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eThe objective of this study is to determine the existence of various cytokines in the serum of patients affected by immune-mediated hearing loss, preserved in a bio bank, in order to establish their role in the etiopathogenesis of the immune response (and posterior use as diagnostic markers) as well as to search for possible new treatment alternatives.\u003c/p\u003e"},{"header":"MATERIALS AND METHODS","content":"\u003cp\u003eWe conducted a retrospective study involving 70 patients diagnosed with immune-mediated hearing loss, alongside 54 healthy adult controls with no prior history of hearing impairment or autoimmune/autoinflammatory disorders.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eINCLUSION CRITERIA\u003c/h2\u003e \u003cp\u003eImmunomediated inner ear disease (IMIED) patients included in the present study fulfilled the inclusion criteria previously reported.\u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e Patients had a diagnosis of IMIED if they had an episode of presence of sensorineural hearing loss manifested as a greater progress at any frequency and audiometric evidence of progression in at least one ear manifested as a threshold shift greater than 15 dB at any frequency or 10 dB at two or more frequencies on serial audiograms in \u0026gt;\u0026thinsp;3 days but less than 3 months apart.\u003c/p\u003e \u003cp\u003eIf the hearing loss developed in \u0026lt;\u0026thinsp;3 days, the patient was included if they presented a suspected immune-mediated profile: two or more episodes in one year and/or bilateral onset, presence of antinuclear autoantibodies (ANAs)\u0026thinsp;\u0026gt;\u0026thinsp;1/160, altered T-cell immunophenotype, response to corticosteroids (oral or intratympanic) in each episode (except in cases with profound hearing loss), possible association with systemic autoimmune diseases.\u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e,\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eHealthy controls were adults without history of hearing loss and autoimmune/autoinflammatory disorders.\u003c/p\u003e \u003cp\u003ePatients were treated with 60mg of oral prednisone or methylprednisolone for 3\u0026ndash;4 weeks followed by a variable taper (2 months taper for rapidly progressive SNHL and 1\u0026ndash;2 weeks for sudden SNHL). The pure-tone average included frequencies: 250, 500, 1000, 2000, 4000 Hz to determine corticosteroid response.\u003c/p\u003e \u003cp\u003eCriteria of a response to corticosteroid therapy included the following:\u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e1. For patients with rapidly progressive sensorineural hearing loss (\u0026gt;\u0026thinsp;3 days to \u0026lt;\u0026thinsp;90 days), improvement in the pure-tone average of 5 dB and a 15 dB improvement in 1 frequency or 10 dB at 2 frequencies, for at least 4 weeks while receiving the therapy.\u003csup\u003e\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u003c/sup\u003e Weekly audiograms were obtained during a 2-month period.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003e2. For patients with sudden sensorineural hearing loss (\u0026lt;\u0026thinsp;3 days): \u003csup\u003e\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u003c/sup\u003e improvement in the final pure-tone average of more than 10dB. Serial audiograms were obtained for 1 year.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e \u003cp\u003e Samples and pertinent patient data utilized in this study were sourced from our Center's Biobank, adhering to standard operating protocols and with requisite approval from both Ethics and Scientific Committees.\u003c/p\u003e \u003cp\u003eThe samples from these patients were collected at the time of diagnosis after obtaining informed consent. Frozen serum samples were thawed prior to analysis, and none of them underwent more than two freeze-thaw cycles before analysis, following the instructions provided in the kit. The cytokines levels in serum were assessed using a Magnetic Luminex Assay kit (ME-HCYT-60K-06, Merck Millipore). A panel of six cytokines (IFN- γ, IL-1 β, IL-6, IL-10, IL-17A y TNF- α) was selected for analysis and measured in a Luminex 200 analyzer instrument according to the manufacturer\u0026rsquo;s protocol. Luminex data were analyzed using Xponent software.\u003c/p\u003e \u003cp\u003e All procedures were conducted in compliance with the applicable regulations and guidelines of the Drug Research Ethics Committee at Puerta de Hierro Majadahonda University Hospital. The study received approval under protocol number 07/2024. All procedures were compliant with the ethical principles for medical research involving human subjects as outlined in the WMA Declaration of Helsinki.\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cp\u003eClinical Characteristics (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e):\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eDemographics and Clinical presentation\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ePatients\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eControls\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSEX\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e29\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAGE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAverage age\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e42.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e41.6\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eCLINIC\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSudden sensorineural hearing loss\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e38 (54.28%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProgressive hearing loss\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e25 (35.72%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eFluctuating hearing loss\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7 (10%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThis study encompassed a cohort of 70 patients diagnosed with IMIED. Among them, 35 (50%) were male, and 35 (50%) were female, with an average age of 42.1 years. The control group consisted of 54 specimens, comprising 29 (54%) females and 25 (46%) males, with an average age of 41.6 years.\u003c/p\u003e \u003cp\u003eBilateral hearing loss was observed in 21 patients (30%). Sudden sensorineural hearing loss was the most frequent clinical form of presentation (38 cases, 54.2%).\u003c/p\u003e \u003cp\u003eIL-10 cytokine levels were found to be elevated in patients diagnosed with IMIED compared to the control group, although the results are not statistically significant. Conversely, the levels of other cytokines were higher in control patients than in those with IMIED with no statistically significant differences except for IL-6 (p\u0026thinsp;=\u0026thinsp;0.025). (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e)\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCytokines results\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCytokine\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eCytokine type\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003ep50 control\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep50 patients\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ep75 control\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003ep75 patients\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eStatistical test\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003ep-valor\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eINF-γ\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePro-inflammatory\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e9.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMann-Whitney U\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIL-1β\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePro-inflammatory\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMann-Whitney U\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIL-6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePro-inflammatory\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.97\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e3.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMann-Whitney U\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIL-10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAnti-inflammatory\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMann-Whitney U\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIL-17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePro-inflammatory\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e1.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMann-Whitney U\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTNF-α\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePro-inflammatory\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e26\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e38\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003eMann-Whitney U\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e\u0026gt;\u0026thinsp;0.05\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThe diagnosis of idiopathic sudden sensorineural hearing loss (ISSNHL) still heavily relies on clinical evaluation in many cases, with only limited markers available to assist and confirm the diagnosis. The immunopathological mechanisms underlying this condition remain elusive. Due to the impracticality of biopsying the inner ear, which would result in its destruction, alternative investigative methods have been explored, such as MRI studies utilizing intratympanic gadolinium \u003csup\u003e\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u003c/sup\u003e. However, the ultimate pathophysiological or immunopathological mechanism has yet to be fully elucidated.\u003c/p\u003e \u003cp\u003eIn recent years, several articles have proposed the correlation and potential involvement of certain interleukins as immunopathological mechanisms in this condition \u003csup\u003e\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u003c/sup\u003e. However, currently, there are no cytokines identified to definitively support the diagnosis of this disease. The findings across published studies remain controversial. (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e)\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePrevious research on the correlation and potential involvement of interleukins in immune mediated inner ear disease\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eStudy\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMolecules\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eResults\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmor Dorado JC et al\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHLA DRA1\u0026lowast;04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMarker of autoimmune middle ear disease, worse prognosis associated.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePsillas G. et al\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHLA DRA1\u0026lowast;04\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003efavorable response to treatment (non statistically significant)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSW Gorthey et al\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIL-6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eElevated IL6 levels are associated with a worse response to treatment.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMoleon MDC et al\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eIL-1β, CCL3, CCL4, CXCL1, CCL22, CCL8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eCytokine levels do not allow distinguishing between late-onset or early-onset Meniere's disease.\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThere have been studies examining the association between HLA and IMIED, although they are limited in number \u003csup\u003e\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e\u003c/sup\u003e. Certain HLA types, such as HLA DRB104, have been identified as potential markers for autoimmune inner ear diseases \u003csup\u003e\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e\u003c/sup\u003e. HLA DRB104 has demonstrated prognostic significance in patients experiencing sudden sensorineural hearing loss, being linked to a poorer treatment outcome.\u003c/p\u003e \u003cp\u003eHowever, in the study conducted by Psillas et al. \u003csup\u003e\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e\u003c/sup\u003e, was examined the correlation between HLA and response to corticosteroid treatment. In this instance, a favorable response to treatment was observed, although the outcome did not reach statistical significance.\u003c/p\u003e \u003cp\u003eGorthey et al. conducted an analysis investigating the correlation between IL-6 levels and the efficacy of corticosteroid treatment in individuals with autoimmune-related hearing loss. Their findings demonstrated that heightened IL-6 levels corresponded with a reduced response to treatment, offering significant insights for timely therapeutic interventions in such patients \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. In our current study, IL-6 levels were observed to be higher in controls compared to patients. However, it is noteworthy that all patients exhibited a positive response to treatment, as the diagnosis was established in the early stages, and positive treatment response was a criterion for diagnosis.\u003c/p\u003e \u003cp\u003eIn 2020, Moleon et al. employed multiplex technology to quantify various cytokine levels in Meniere's disease, with the objective of delineating the cytokine profile based on the age of disease onset. The study did not yield statistically significant differences overall. However, when stratifying by gender, notable distinctions emerged: levels of ILB were found to be higher in men than in women during early onset, whereas levels of CXCL1 were higher in women compared to men.\u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eIn the present study, elevated levels of IL-10 were observed in individuals with autoimmune hearing loss. Moreover, given the favorable response to corticosteroid therapy exhibited by the study participants, this biomarker may serve as a predictive indicator for treatment response. IL-10 serves as a crucial anti-inflammatory cytokine with significant effects on antigen-presenting cells. It operates by impeding the synthesis of proinflammatory cytokines such as IL1-β, TNF-α, and IL-6, along with Th2 cell-derived cytokines like IL-4 and IL-5. Additionally, IL-10 exerts a direct regulatory influence on T cell differentiation by suppressing IL-2 and IFN-γ.\u003csup\u003e\u003cspan additionalcitationids=\"CR26 CR27\" citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThis observation offers a potential explanation for the elevated IL-10 levels in our patients alongside the comparatively lower levels of other analyzed cytokines in comparison to control subjects. Likewise, IL-10 also demonstrates higher expression in patients with idiopathic ulcerative colitis during remission. This upregulation is attributed to its anti-inflammatory properties within the intestine, where it acts to inhibit antigen presentation and the subsequent release of proinflammatory cytokines.\u003csup\u003e\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e\u003c/sup\u003e. In this context, the administration of recombinant IL-10 and gelatin microspheres incorporating IL-10 in ulcerative colitis has been suggested.\u003csup\u003e\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e \u003cp\u003eBased on this finding, we propose a novel treatment strategy with IL-10 (transtympanic or systemically administrated) for corticosteroids non-responder IMIED patients.\u003c/p\u003e \u003cp\u003eSince the inner ear lacks a lymphatic system the endolymphatic sac could potentially function as a mucosa-associated lymphoid tissue (MALT) organ within the inner ear \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e\u003c/sup\u003e, transtympanic injection of IL-10 may represent a new therapeutic strategy in IMIED patients.\u003c/p\u003e \u003cp\u003eRecently, Xinyuan Tan et al.\u003csup\u003e\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u003c/sup\u003e delve into the significance of the interplay between inflammatory and anti-inflammatory cytokines in the pathogenesis of sudden hearing loss, as well as our proposed theory on autoimmune hearing loss. This elucidates the favorable response to corticosteroid treatment, attributed to their anti-inflammatory properties.\u003c/p\u003e \u003cp\u003eThe findings from the experimental study conducted in mice by Zhou et al. provide support for the theory behind this novel treatment approach. Their results indicate that a deficiency in IL-10 worsens hearing loss, while the exogenous administration of IL-10 enhances hearing function. These outcomes underscore the pivotal role of IL-10 as a key regulator of inflammation in vivo. \u003csup\u003e\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eDue to the extended duration of IMIED, which may lead to hearing fluctuations unresponsive to corticosteroids, there's a need to explore the role of cytokines in later stages. This prompts our ongoing research into the rationale behind IL-10 treatment for non-responders.\u003c/p\u003e"},{"header":"CONCLUSIONS","content":"\u003cp\u003eIn recent years, studies have aimed to assess cytokine levels in patients with autoimmune hearing loss, with the goal of elucidating the etiopathogenesis of this condition. However, further research is required to fully understand the underlying mechanisms. The elevated levels of IL-10 observed in our patients may account for the absence of elevated levels of other analyzed cytokines, owing to its anti-inflammatory properties.\u003c/p\u003e \u003cp\u003eDespite the limitations inherent in our retrospective study design, the substantial number of participants enables us to support the hypothesis regarding the potential role of IL-10 administration in patients who do not respond to corticosteroid treatment. Our findings suggest a promising avenue for further investigation into the efficacy of IL-10 therapy in this subset of patients.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eASM: data collecting, writing and editing the first draftNAC: data collectingAR: biostatistical analysisJMVM: manuscript review, corresponding authorAJSL: data analysisSRG: data analysisJRGB: research coordinator, final approvalAcknowledgments \u0026ldquo;The authors wish to thank the donors, and the Hospital Universitario Puerta de Hierro Majadahonda (HUPHM)/Instituto de Investigaci\u0026oacute;n Sanitaria Puerta de Hierro-Segovia de Arana (IDIPHISA) Biobank (Carlos III Health Institute Biobanks and Biomodels Platform \u0026ndash; PT23/00015) for the human specimens used in this study\u0026rdquo;.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eThe authors wish to thank the donors, and the Hospital Universitario Puerta de Hierro Majadahonda (HUPHM)/Instituto de Investigaci\u0026oacute;n Sanitaria Puerta de Hierro-Segovia de Arana (IDIPHISA) Biobank (Carlos III Health Institute Biobanks and Biomodels Platform \u0026ndash; PT23/00015) for the human specimens used in this study.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eThe data that support the findings of this study are not openly available due to reasons of sensitivity and are available from the corresponding author upon reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eMcCabe, B. F. Autoimmune sensorineural hearing loss. Ann. Otol. Rhinol. Laryngol. 88, 585\u0026ndash;589 (1979).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarc\u0026iacute;a-Berrocal, J. R. \u003cem\u003eet al.\u003c/em\u003e Otosyphilis mimics immune disorders of the inner ear. Acta Otolaryngol. (Stockh.) 126, 679\u0026ndash;684 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHerrera, M., Garc\u0026iacute;a Berrocal, J. R., Garc\u0026iacute;a Arum\u0026iacute;, A., Lavilla, M. J. \u0026amp; Plaza, G. Actualizaci\u0026oacute;n del consenso sobre el diagn\u0026oacute;stico y tratamiento de la sordera s\u0026uacute;bita idiop\u0026aacute;tica. Acta Otorrinolaringol\u0026oacute;gica Esp. 70, 290\u0026ndash;300 (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoleon, M.-D.-C. \u003cem\u003eet al.\u003c/em\u003e Clinical and Cytokine Profile in Patients with Early and Late Onset Meniere Disease. J. Clin. Med. 10, 4052 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLobo, D., L\u0026oacute;pez, F. G., Garc\u0026iacute;a-Berrocal, J. R. \u0026amp; Ram\u0026iacute;rez-Camacho, R. Diagnostic tests for immunomediated hearing loss: a systematic review. J. Laryngol. Otol. 122, 564\u0026ndash;573 (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarc\u0026iacute;a-Berrocal, J. R. \u003cem\u003eet al.\u003c/em\u003e Alternatives to systemic steroid therapy for refractory immune-mediated inner ear disease: A physiopathologic approach. \u003cem\u003eEur. Arch. Oto-Rhino-Laryngol. Off. J. Eur. Fed. Oto-Rhino-Laryngol. Soc. EUFOS Affil. Ger. Soc. Oto-Rhino-Laryngol. - Head Neck Surg.\u003c/em\u003e 263, 977\u0026ndash;982 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDavid R Lobo, J. R. G., a-Berrocal, R. R. \u0026amp; rez-Camacho. New prospects in the diagnosis and treatment of immune-mediated inner ear disease. World J. Methodol. 4, 91\u0026ndash;98 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarcia-Berrocal, J. R. \u003cem\u003eet al.\u003c/em\u003e Deficiency of Naive T Cells in Patients With Sudden Deafness. Arch. Otolaryngol. - Head Neck Surg. 123, 712\u0026ndash;717 (1997).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLobo, D., Garc\u0026iacute;a-Berrocal, J. R., Trinidad, A., Verdaguer, J. M. \u0026amp; Ramirez-Camacho, R. Review of the biologic agents used for immune-mediated inner ear disease. Acta Otorrinolaringol. Esp. (2012) doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.otorri.2012.04.008\u003c/span\u003e\u003cspan address=\"10.1016/j.otorri.2012.04.008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarc\u0026iacute;a Berrocal, J. R. \u0026amp; Ram\u0026iacute;rez-Camacho, R. Immune response and immunopathology of the inner ear: an update. J. Laryngol. Otol. 114, 101\u0026ndash;107 (2000).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHashimoto, S., Billings, P., Harris, J. P., Firestein, G. S. \u0026amp; Keithley, E. M. Innate immunity contributes to cochlear adaptive immune responses. Audiol. Neurootol. 10, 35\u0026ndash;43 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGorthey, S. W., Pathak, S. \u0026amp; Vambutas, A. The Correlation of Clinical Corticosteroid Responsiveness With Expression of IL-6 in Peripheral Blood Immune Cells (PBMC) in Patients With Autoimmune Inner Ear Disease (AIED). Otol. Neurotol. 42, 1422\u0026ndash;1428 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLobo, D., Trinidad, A., Garc\u0026iacute;a-Berrocal, J. R., Verdaguer, J. M. \u0026amp; Ram\u0026iacute;rez-Camacho, R. TNFalpha blockers do not improve the hearing recovery obtained with glucocorticoid therapy in an autoimmune experimental labyrinthitis. \u003cem\u003eEur. Arch. Oto-Rhino-Laryngol. Off. J. Eur. Fed. Oto-Rhino-Laryngol. Soc. EUFOS Affil. Ger. Soc. Oto-Rhino-Laryngol. - Head Neck Surg.\u003c/em\u003e 263, 622\u0026ndash;626 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLorenz, R. R. \u003cem\u003eet al.\u003c/em\u003e Interferon-g production to inner ear antigens by T cells from patients with autoimmune sensorineural hearing loss. J. Neuroimmunol. (2002).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVambutas, A. \u003cem\u003eet al.\u003c/em\u003e Early efficacy trial of anakinra in corticosteroid-resistant autoimmune inner ear disease. J. Clin. Invest. 124, 4115\u0026ndash;4122 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMoscicki, R. A. \u003cem\u003eet al.\u003c/em\u003e Serum antibody to inner ear proteins in patients with progressive hearing loss. Correlation with disease activity and response to corticosteroid treatment. JAMA 272, 611\u0026ndash;616 (1994).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBerrocal, J. R. G. \u0026amp; Ram\u0026iacute;rez-Camacho, R. Sudden sensorineural hearing loss: supporting the immunologic theory. Ann. Otol. Rhinol. Laryngol. 111, 989\u0026ndash;997 (2002).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarc\u0026iacute;a-Berrocal, J. R. \u003cem\u003eet al.\u003c/em\u003e Sudden presentation of immune-mediated inner ear disease: characterization and acceptance of a cochleovestibular dysfunction. J. Laryngol. Otol. 117, 775\u0026ndash;779 (2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNiparko, J. K. \u003cem\u003eet al.\u003c/em\u003e Serial Audiometry in a Clinical Trial of AIED Treatment. Otol. Neurotol. 26, 908\u0026ndash;917 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLobo, D., Tu\u0026ntilde;\u0026oacute;n, M., Villarreal, I., Brea, B. \u0026amp; Garc\u0026iacute;a-Berrocal, J. R. Intratympanic gadolinium magnetic resonance imaging supports the role of endolymphatic hydrops in the pathogenesis of immune-mediated inner-ear disease. J. Laryngol. Otol. 132, 554\u0026ndash;559 (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHajas, A. \u003cem\u003eet al.\u003c/em\u003e Sensorineural Hearing Loss in Patients with Mixed Connective Tissue Disease: Immunological Markers and Cytokine Levels. J. Rheumatol. 36, 1930\u0026ndash;1936 (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGarc\u0026iacute;a-Berrocal, J. R. \u003cem\u003eet al.\u003c/em\u003e Deficiency of naive T cells in patients with sudden deafness. Arch. Otolaryngol. Head Neck Surg. 123, 712\u0026ndash;717 (1997).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmor-Dorado, J. C., Paco, L., Martin, J., Lopez-Nevot, M. A. \u0026amp; Gonzalez-Gay, M. A. Human leukocyte antigen-DQB1 and -DRB1 associations in patients with idiopathic sudden sensorineural hearing loss from a defined population of Northwest Spain. Acta Otolaryngol. (Stockh.) 125, 1277\u0026ndash;1282 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePsillas, G., Binos, P., Dimas, G. G., Daniilidis, M. \u0026amp; Constantinidis, J. Human Leukocyte Antigen (HLA) Influence on Prognosis of Autoimmune Hearing Loss. Audiol. Res. 11, 31\u0026ndash;37 (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEbert, E. C. IL-10 enhances IL-2-induced proliferation and cytotoxicity by human intestinal lymphocytes. Clin. Exp. Immunol. 119, 426\u0026ndash;432 (2000).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFonseca-Camarillo, G., Furuzawa-Carballeda, J., Mart\u0026iacute;nez-Ben\u0026iacute;tez, B., Barreto-Z\u0026uacute;\u0026ntilde;iga, R. \u0026amp; Yamamoto-Furusho, K. [Intelukin-10 expression with immunorreglatory function in the mucosa of patients with ulcerative colitis]. Rev. Gastroenterol. Mex. 76, 113\u0026ndash;119 (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLi, M.-C. IL-10 and its related cytokines for treatment of inflammatory bowel disease. World J. Gastroenterol. 10, 620 (2004).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eStaples, K. J., Bergmann, M., Barnes, P. J. \u0026amp; Newton, R. Stimulus-Specific Inhibition of IL-5 by cAMP-Elevating Agents and IL-10 Reveals Differential Mechanisms of Action. Biochem. Biophys. Res. Commun. 273, 811\u0026ndash;815 (2000).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTan, X. \u003cem\u003eet al.\u003c/em\u003e Relevant Research of Inflammatory Cytokines Spectrum in Peripheral Blood of Sudden Hearing Loss. The Laryngoscope lary.31276 (2024) doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1002/lary.31276\u003c/span\u003e\u003cspan address=\"10.1002/lary.31276\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhou, B. \u003cem\u003eet al.\u003c/em\u003e Experimental autoimmune hearing loss is exacerbated in IL-10-deficient mice and reversed by IL-10 gene transfer. Gene Ther. 19, 228\u0026ndash;235 (2012).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Immune-mediated hearing loss, Cytokines, Corticosteroid therapy, IL-10, Autoimmune inner ear disease (AIED)","lastPublishedDoi":"10.21203/rs.3.rs-4622777/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4622777/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose:\u003c/strong\u003e\u003cbr\u003e\nDiagnosing immune-mediated hearing loss is challenging due to diverse clinical presentations and lack of specific markers. This study assesses cytokine levels in these patients to identify potential diagnostic markers and new treatments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMaterials and Methods:\u003c/strong\u003e\u003cbr\u003e\nA retrospective analysis of 70 early-stage immune-mediated hearing loss patients was conducted. Inclusion criteria included clinical presentation, immunological work-up alterations, corticosteroid response, and comparison with healthy controls. Serum levels of IFN gamma, IL-1Beta, IL-6, IL-10, IL-17a, and TNFalfa were measured using a Magnetic Luminex Assay kit.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e\u003cbr\u003e\nThe cohort, with an average age of 42.1 years, showed bilateral hearing loss in 30% of cases, predominantly as sudden sensorineural hearing loss (54.2%). Among cytokines analyzed, only IL-10 levels were elevated compared to controls.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDiscussion:\u003c/strong\u003e\u003cbr\u003e\nDiagnosis relies on clinical evaluation due to limited diagnostic markers. Previous studies on cytokine involvement had conflicting results. Elevated IL-10 levels suggest a role in pathogenesis and treatment response, warranting further investigation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e\u003cbr\u003e\nIL-10 therapy is proposed for corticosteroid non-responders, offering a promising research direction. Identifying diagnostic markers and understanding immune-related causes are crucial for improving outcomes in immune-mediated hearing loss, supporting ongoing research efforts.\u003c/p\u003e","manuscriptTitle":"Cytokine analysis may support new therapeutic strategies for immune-mediated hearing loss.","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-07-22 20:50:14","doi":"10.21203/rs.3.rs-4622777/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"editorInvitedReview","content":"","date":"2024-08-20T10:27:26+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-08-19T21:16:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"297277462626451829085803597473579048008","date":"2024-08-02T08:18:10+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"7356248611159048192540641527877149268","date":"2024-08-02T06:46:12+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-01T15:51:57+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-01T15:46:32+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2024-06-27T17:00:12+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-06-27T05:08:11+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2024-06-22T17:07:57+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"004f820f-f622-4a93-8c07-1283febca4e3","owner":[],"postedDate":"July 22nd, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":34563717,"name":"Biological sciences/Immunology/Applied immunology"},{"id":34563718,"name":"Biological sciences/Immunology/Cytokines"},{"id":34563719,"name":"Biological sciences/Immunology/Immunological disorders"},{"id":34563720,"name":"Biological sciences/Immunology/Neuroimmunology"},{"id":34563721,"name":"Health sciences/Diseases/Immunological disorders/Autoimmune diseases"}],"tags":[],"updatedAt":"2024-07-22T20:50:15+00:00","versionOfRecord":[],"versionCreatedAt":"2024-07-22 20:50:14","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4622777","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4622777","identity":"rs-4622777","version":["v1"]},"buildId":"_2-kVJe1T_tPrBINL-cwx","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}