Antigen presentation by Toxoplasma-infected cells: antigen entry through cell membrane fusion

The antigen presentation of cells infected by Toxoplasma gondii (T.g.) for MHC class-I-restricted cytotoxic T cells (CTL) was blocked by treating the T.g.-infected cells with the fungal antibiotic brefeldin A. Electron microscopic analysis of T.g.-infected murine and human-antigen-presenting cells (APC) prepared by the combination of quick-freezing with the deep-etching or freeze-substitution methods revealed that the outer membrane of T.g. fused with some parts of the vacuolar membranes of APC to build up channel-like structures. Work station and cytofluorometry studies detected 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein (BCECF) in cells infected by BCECF-labeled T.g., which strongly suggested the direct entry into cells of BCECF derived from T.g. These findings provide evidence that T.g. antigens enter the cytoplasm of APC through cell membrane fusion of T.g. and T.g.-infected APC, and that T.g.-infected APC present T.g. antigen for MHC class-I-restricted CTL through the endogenous pathway of antigen presentation.