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J. Exp. Med.,
Volume 186, Number 9, November 3, 1997 1557-1565
By
From the * Institute for Specific Prophylaxis and Tropical Medicine, A-1095 Vienna, Austria; A panel of monoclonal antibodies was raised from mice immunized with a membrane preparation from Entamoeba histolytica, the pathogenic species causing invasive amebiasis in humans.
Antibody EH5 gave a polydisperse band in immunoblots from membrane preparations from
different E. histolytica strains, and a much weaker signal from two strains of the nonpathogenic
species Entamoeba dispar. Although the exact chemical structure of the EH5 antigen is not yet
known, the ability of the antigen to be metabolically radiolabeled with [32P]phosphate or
[3H]glucose, its sensitivity to digestion by mild acid and phosphatidylinositol-specific phospholipase C, and its specific extraction from E. histolytica trophozoites by a method used to prepare
lipophosphoglycans from Leishmania showed that it could be classified as an amebal lipophosphoglycan. Confocal immunofluorescence and immunogold labeling of trophozoites localized
the antigen on the outer face of the plasma membrane and on the inner face of internal vesicle
membranes. Antibody EH5 strongly agglutinated amebas in a similar way to concanavalin A
(Con A), and Con A bound to immunoaffinity-purified EH5 antigen. Therefore, surface lipophosphoglycans may play an important role in the preferential agglutination of pathogenic
amebas by Con A. The protective ability of antibody EH5 was tested in a passive immunization
experiment in a severe combined immunodeficient (SCID) mouse model. Intrahepatic challenge of animals after administration of an isotype-matched control antibody or without treatment led to the development of a liver abscess in all cases, whereas 11 out of 12 animals immunized
with the EH5 antibody developed no liver abscess. Our results demonstrate the importance
and, for the first time, the protective capacity of glycan antigens on the surface of the amebas.
Institute of General and Experimental Pathology, A-1090 Vienna, Austria; § Department of Medicine
and Molecular Microbiology, Washington University School of Medicine, St. Louis, Missouri 63110; Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur, 75724 Paris, France; and ¶ Station
Centrale de Microscopie Electronique, Institut Pasteur, 75724 Paris, France
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