Production and characterization of monoclonal antibodies to Acanthamoeba castellanii and their application for detection of pathogenic Acanthamoeba spp

Abstract

Fourteen monoclonal antibodies (mAbs) were produced against a strain of Acanthamoeba castellanii isolated from a human cornea. The reactivity of the mAbs to reference strains of Acanthamoeba was examined by an indirect fluorescence antibody test (IFA) and Western immunoblot analysis. Nine mAbs reacted specifically with a known pathogenic reference strain of A. castellanii, but not with a non-pathogenic strain or other Acanthamoeba spp. The antigen recognized by these mAbs had a molecular mass of 17 kDa. The remaining five mAbs reacted with A. castellanii and A. polyphaga, members of group II (Pussard and Pons) but not with A. astronyxis (group I) or A. culbertsoni (group III). Western immunoblot analysis revealed that the latter mAbs stained many protein bands ranging from 30 to 150 kDa. None of the 14 mAbs reacted with Naegleria gruberi, N. fowleri, or Entamoeba histolytica. These observations suggest that an antigen common in group II as well as a pathogenic A. castellanii-specific antigen are present. Slot blot reactivity was comparable to the IFA. Under certain circumstances, therefore, slot blot analysis with a panel of mAbs should be helpful in the detection of keratitis-producing strains of Acanthamoeba.