Toxoplasma gondii: immunocytochemistry of four immunodominant antigens with monoclonal antibodies.
Four monoclonal antibodies in which diagnostic usefulness has been observed, concerning congenital, acquired, and reactivated toxoplasmosis, were raised against Toxoplasma gondii tachyzoïtes in order to localize immunodominant antigens. On immunoblots, it appears that McAb IV47, McAB GII9, McAb II38, and McAb IE10 identify families of proteins with estimated molecular weights of 28-30 kDa, 30 kDa, 45-50 kDa, and 66-70 kDa, respectively. By immunogold preembedding techniques one can observe an homogeneous labeling of the outer pellicle of the tachyzoïtes with the McAb GII9 and IV47 and a light labeling with the McAb II38 and IE10. The three-dimensional observation of cell surface antigens is performed by applying a modified metal extraction replica method, i.e., A plasma polymerization method of glow discharge by Tanaka (1979). By immunogold preembedding techniques [with saponin permeabilization (0.1%)], and by immunogold postembedding techniques, a labeling of the rhoptries is observed with McAb GII9 and McAb IV47 but essentially all label is found with McAb II38 and IE10. With McAb GII9 a uniform labeling is observed on the cell surface. By immunoenzymatic techniques (peroxidase) a cell surface labeling is observed with the four McAb. Intracellular Toxoplasma, the outer pellicle, and the vesicles of the network (elaborated by Toxoplasma in parasitophorous vacuole) are also labeled with McAb IE10. These results indicate that McAb GII9 recognizes antigens of the antigen family (P 30) located on the cell surface and in the rhoptries. The antigen recognized by McAb IV47 is essentially located on and beneath the Toxoplasma cell surface membrane, and McAb II38 and IE10 identify preferentially rhoptry proteins.
Bonhomme
A
A
Unité INSERM U.314, Hôpital Maison Blanche, Reims, France.
Boulanger
F
F
Bharadwaj
L M
LM
Puygauthier-Toubas
D
D
Bonhomme
P
P
Pluot
M
M
Pinon
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan / Q000032:analysis*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D007150:Immunohistochemistry; D016253:Microscopy, Immunoelectron; D014122:Toxoplasma / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan