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Hassan Alizadeh, Sudha Neelam, Jerry Y. Niederkorn; Effect of Immunization with the Mannose-Induced Acanthamoeba Protein and Acanthamoeba Plasminogen Activator in Mitigating Acanthamoeba Keratitis. Invest. Ophthalmol. Vis. Sci. 2007;48(12):5597-5604. doi: 10.1167/iovs.07-0407.
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purpose. The mannose-induced cytopathic protein (MIP-133) and Acanthamoeba plasminogen activator (aPA) play key roles in the pathogenesis of Acanthamoeba keratitis by inducing a cytopathic effect on the corneal epithelial and stromal cells and by production of proteolytic enzymes that facilitate the invasion of trophozoites through the basement membrane. The goal of the present study was to gain insight into the pathogenicity of Acanthamoeba infection as well as to determine whether oral immunization with aPA and MIP-133 produce an additive protection against Acanthamoeba keratitis.
methods. MIP-133 and aPA were isolated by chromatography. The purity of the concentrated MIP-133 and aPA was confirmed by SDS-PAGE and fibrinolytic activity, respectively. aPA activity of Acanthamoeba cultures was quantitated by radial diffusion in fibrin-agarose gel. The capacity of aPA and MIP-133 to induce cytolysis of corneal epithelial cells was tested in vitro. Chinese hamsters were orally immunized with four weekly doses of aPA or MIP-133 conjugated with cholera toxin. The animals were immunized before infection to determine the prophylactic effect of oral immunization. The therapeutic effect of oral immunization with aPA and MIP-133 was determined after corneal infection had been established. The animals were then infected via Acanthamoeba castellanii–laden contact lenses.
results. aPA was characterized in pathogenic and nonpathogenic strains of Acanthamoeba spp. Oral immunization with MIP-133 before and after infection with Acanthamoeba significantly reduced the severity of corneal infection which includes infiltration and ulceration (P < 0.05) and shortened the duration of the disease. Immunization with aPA alone did not significantly affect the course of disease (P > 0.05).
conclusions. These data suggest that once trophozoites invade the cornea, MIP-133 production is necessary to initiate corneal disease and plays an important role in the subsequent steps of the pathogenic cascade of Acanthamoeba keratitis.
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