Detection of candidate antigen of Acanthamoeba castellanii practicable for immunological examination of Amoebic Keratitis

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We investigated cytopathic effect of Acanthamoeba T2, T4 and T7 genotypes on human corneal epithelial cell (HCE-T). Cytopathic effects were observed not only in T4 genotype but also in T2 genotypes that classified as nonpathogenic species by 18SrRNA genotyping. These results suggested the discrepancy between 18sRNA typing and actual pathogenicity of Acanthamoeba species. In addition, we found that protease inhibitor did not inhibit cytopathic effect of Acanthameoba on HCE-T cell, and after challenging Acanthamoeba to rabbit kidney 13 cell induced interferon via interferon-sensitive response element. These results suggested the existence of novel mechanism of Acanthamoeba keratitis. To investigate antigen which might be useful for diagnosis of amoebic keratitis, we constructed cDNA library of A. castellanii and screened immunologically using polyclonal rabbit antibody against A. castellanii soluble antigen. However, in spite of our expectation, we could not obtain positive clones.