Abstract Background Neuromyelitis Optica (NMO) is an autoimmune disease of the Central Nervous System (CNS) mediated by Th17 cells and antibody response to the water channel protein Aquaporin 4 (AQP4), leading to chronic damage of the optic nerves and spinal cord. NMO animal models are substandard inducing disease inconsistently and weakly. Objectives Characterize a new NMO mouse model, which best represents the human disease in terms of incidence and chronicity of the disease, severity of symptoms and underlying molecular mechanisms. Methods C57BL/6 (WT) and IFN-γ receptor knockout (R-KO) mice were immunized subcutaneously with the AQP4201–220 peptide in Complete Freund Adjuvant and treated with pertussis toxin at day 0 and +2. WT were treated weakly with neutralizing anti-IFN-γ antibodies. mRNA expression and flow cytometry analysis of infiltrating CNS cells were performed. CNS histopathology was determined by immunofluorescence. In vivo B-cell depletion was made using anti-CD20 antibodies. Results WT mice treated with anti-IFN-γ and RKO mice showed ascendant paralysis starting from the tail to complete hind limb paralysis. Mice showed increased disease incidence and symptoms were more severe and chronic. CNS IL-17 expressing T cells and Th17 associated genes were upregulated in the absence of IFN-γ. CSN histological analysis showed an increased presence of T and B cells in lesion sites, associated with astrocytes and myelin loss. B-cell depletion in AQP4 primed RKO mice, diminished the incidence and severity of the disease confirming B cell pathogenicity. Conclusions AQP4 mouse model is strictly regulated by IFN-γ and its signaling, which control the disease progression and severity, by restraining pathogenic Th17 and B cells. Supported by grant from NIH (R21 Grant 12878760)