The long term objective of this project is to develop a new gene vaccination therapy for the autoimrnune hearing loss and Meniere's disease in humans. The immediate goal of this study is to induce autoimmune hearing loss induced by beta,-tubulin in mice to quantify immune responses and to modulate the hearing loss by immunologic methods.
The specific aims are as follows. 1. To induce autoimmune hearing loss in mice strains -using varying concentrations of Tubulin. 2. To quantify the Immune response by analyzing Thl and Th2 cell functions. 3. To modulate tubulin-induced hearing loss by immunological means. 4. To determine if oral feeding of tubulin would prevent or improve hearing loss. This will eventually enable us to develop a new therapeutics: a naked DNA vaccine for hearing loss; Meniere's disease and autoimmune sensorineural hearing loss. These goals will be achieved as follows. 1. Mice w ill be immunized with varying concentration of tubulin to enhance the incidence of hearing loss. 2. The immune response will be quantified by a. By measuring IgG classes of immunoglobulin : circulating (by ELISA), and detect tubulin specific IgG classes of antibodies in the ear tissues by immunohistochemistry. b. By measuring cytokines IL-2, IFNg]amma, IL-4, IL-10, TGF-beta following tubulin immunization and induction of hearing loss. 3. Tubulin induced hearing loss will be modulated by following immunologic methods. a. By administering dendritic cells engineered to express tubulin (plasmid transfection) b. By administering plasmid DNA expressing IFN-gamma to enhance the disease severity. c. By administering plasmid DNA expressing TGF-beta, IL-10 or IL-4 to reduce the disease severity. 4. Oral tolerance study will be carried out by feeding (low dose, medium dose or high dose and cholera toxin B chain linked antigen) of tubulin in mice before or after immunization with tubulin to determine the efficacy of oral tolerance as either preventative measure or therapeutics.
