MORPHOLOGICAL AND IMMUNOLOGICAL STUDIES ON MICROSPORIDIA - ANTIGENE PROPERTIES OF NOSEMA-MESNILI SPORES

Immunological investigation of two sequences of the life cycle (with m ono- and octospores) of presumably one microsporidian species, Nosema mesnili, included: application of a murine anti-monospores serum in an immunofluorescent assay (IFA), electron microscopy (IEM) analysis and Western blotting of antigenes of both spore types; hybridoma producti on and testing of hybridoma clones by IFA. The spore surface, polar fi laments and sporoplasm gave strong positive immune reaction against th e antiserum in IFA test. The antiserum reacted with spore antigens ran ging in molecular weight from 20 to 100 kDA. Four positive hybridoma c lones reacted only with the surface of monospores. Polyclonal antibodi es recognized the inner structures of spores and vegetative stages of both sequences of development as it has been revealed by IEM. Immuniza tion with spores lead to antibody production against the spore wall, p olar filament and sporoplasm. Cloning the hybridoma cells promised to deliver useful antibodies for investigations of the spore morphogenesi s. The strong crossreaction of the antibodies with two developmental s tages of the life cycle provides good evidence in favour of their belo nging to one species.