To characterize extracellular vesicles proteins cargo from the aqueous humor and plasma of patients with ocular toxoplasmosis. Methods: Aqueous humor (AH) and plasma were collected from 6 patients with active ocular toxoplasmosis (OT) and 6 cataract patients (CAT). Extracellular vesicles (EVs) were isolated, and western blotting and mass spectrometry were performed for protein analysis. Results: All plasma samples from OT and CAT patients were positive for tetraspanins CD63 and TSG101 markers. However, only the AH from OT patients were positive for CD63. A total of 67 new proteins not previously reported in EVs were found in the AH and plasma from OT and CAT groups. Among these, 10 and 7 proteins were found only in the samples from CAT and OT groups, respectively. In general, these proteins were involved in immune system activation, retina homeostasis, factors related to infections and retina-associated disease. Conclusion: The distinct protein signatures between OT and CAT patients may be helpful in the differential diagnosis of OT. However, more studies are needed to better understand the role of these proteins in the pathogenesis of OT.