Entamoeba histolytica: localization of a 30-kDa cysteine proteinase using a monoclonal antibody.
We produced a monoclonal antibody against a major cysteine proteinase of 30kDa from trophozoites of Entamoeba histolytica strain HM1:IMSS. The specificity of the monoclonal antibody was confirmed by specific inhibition of azocasein digestion and by electrophoretic analysis, in the presence of sodium dodecyl sulfate or on a substrate gel, of the antigen precipitated by the antibody. Immunofluorescent staining of trophozoites with the monoclonal antibody revealed heterogeneity in the intensity of whole cell fluorescence and subcellular localization of the stain. The latter was also observed in trophozoites, which were stained by conventional immunohistochemical methods, from experimental liver abscesses in hamsters. Ultrastructural analysis showed antigen distributed mainly in clear amorphous zones in the cytoplasm, which were not limited by a visible membrane. Proteinases are translocated from these compartments to phagocytic vacuoles after trophozoites ingest erythrocytes, suggesting that these regions might be a lysosomal equivalent of this primitive eukaryotic cell.
Becker
I
I
Departamento de Medicina Experimental, Facultad de Medicina U.N.A.M., Mexico City D.F., Mexico.
Pérez-Montfort
R
R
Pérez-Torres
A
A
Rondán-Zárate
A
A
Montfort
I
I
Pérez-Tamayo
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000032:analysis* / Q000276:immunology; D004748:Entamoeba histolytica / Q000201:enzymology* / Q000276:immunology; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas; D007150:Immunohistochemistry; D051379:Mice; D016253:Microscopy, Immunoelectron; D011233:Precipitin Tests
D000911:Antibodies, Monoclonal; D003546:Cysteine Endopeptidases