Quantitative immunolocalization of four immunodominant antigens of Toxoplasma gondii.
Ultrastructural localization of four immunodominant antigens of Toxoplasma gondii was investigated quantitatively on thin sections and replicas by an immunogold technique using four monoclonal antibodies (Mab). On immunoblot Mab IV47, GII9, II38 and IE10 identified proteins of 28, 30, 45 and 66-70 kDa, respectively. Use of digital image analyzer and a semi-automatic procedure developed by us, the patterns of label distribution were compared in three cell structures: cell surface, submembrane area and rhoptries. On the whole cell surface, protein P28 and P30 were 2.5 and 4 times more abundant than P66-70 respectively. The protein P28 was essentially concentrated in the submembrane area with a labeling of 195.4 +/- 46.7 gold particles/microns 2 that follows a decreasing gradient from this area to the cell centre. In the rhoptries, all four antigens were detected, P45 and P66-70 being major with a labeling of 97.1 +/- 31.1 gold particles/microns 2 and 155.1 +/- 39.3 gold particles/microns 2 respectively. The results support the hypothesis that rhoptries are the essential site for antigen storage.
Boulanger
F
F
INSERM U 314 E IVb, Reims, France.
Bonhomme
A
A
Lebonvallet
S
S
Bonnet
N
N
Pluot
M
M
Pinon
J M
JM
D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan / Q000276:immunology*; D015153:Blotting, Western; D007091:Image Processing, Computer-Assisted; D016056:Immunodominant Epitopes / Q000032:analysis*; D007150:Immunohistochemistry / Q000379:methods; D008854:Microscopy, Electron; D014122:Toxoplasma / Q000276:immunology / Q000648:ultrastructure*
D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D016056:Immunodominant Epitopes