Background <p>Ménière’s Disease (MD) is a complex vestibular disorder, and its pathological mechanism has not been fully elucidated so far.</p> Objective <p>Compare the positive rates of anti-myelin-associated antibodies between patients with MD and the normal control group, and analyze the clinical characteristics of MD patients with positive anti-myelin-associated antibodies.</p> Methods <p>Between April 2020 and July 2020, a total of 66 patients diagnosed with definite MD and 42 healthy controls were recruited at Shandong Provincial ENT Hospital. A comprehensive auditory-vestibular function assessment was conducted for the MD patients, which included Pure Tone Audiometry, Speech Discrimination Score (SDS), Auditory Brainstem Response, vestibular-evoked myogenic potential (VEMP), caloric testing, video head impulse test, and inner ear Gadolinium MRI, among other examinations. Additionally, indirect immunofluorescence testing was employed to detect anti-myelin-associated antibodies in all participants, followed by a correlation analysis between the detected results and the clinical manifestations. Furthermore, the vestibular end organ of a MD patient who had undergone labyrinthectomy was obtained through surgery for transmission electron microscopy (TEM) examination, in order to observe the ultrastructure of the vestibular nerve and determine the presence of any demyelinating lesions.</p> Results <p>When compared with the control group, the positivity rate of antibodies related to central nervous system (CNS) demyelinating diseases in patients with MD did not exhibit a statistically significant difference (<i>p</i> = 0.217). In contrast, the positivity rate of antibodies associated with peripheral nerve demyelinating diseases showed a highly significant statistical difference (<i>p</i> &lt; 0.001), the positivity rates of anti-sulfatide antibodies and anti-GM1 antibodies in the MD group were significantly higher than those in the control group (<i>p</i> &lt; 0.001). Patients with positive anti-GM1 antibodies were found to be older (<i>p</i> = 0.014), yet they demonstrated higher SDS (<i>p</i> = 0.023), a lower degree of semicircular canal paresis (<i>p</i> = 0.017), and a reduced abnormal rate of cervical VEMP (<i>p</i> = 0.049). Conversely, no significant correlation was observed between the expression of anti-sulfatide antibodies and the clinical symptoms or auditory-vestibular function levels of the patients. TEM of the vestibular nerve tissue obtained from MD patients revealed distinct demyelinating lesions.</p> Conclusions <p>There is a phenomenon of demyelination of the vestibular nerve in patients with MD, and the positive rate of anti-myelin-associated antibodies in the peripheral blood of these patients is significantly higher than that in the control group. Among them, anti-GM1 antibodies may be correlated with auditory and vestibular functions, and they are expected to become a biological marker for MD.</p>