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* Institut für Medizinische Mikrobiologie und Hygiene, Universitätsklinikum Mannheim, Universität Heidelberg, Mannheim, Germany;
Abteilung für Neuropathologie, Klinikum der Universität zu Köln, Köln, Germany;
Department of Biological Sciences, Imperial College of Science, Technology and Medicine, London, United Kingdom; and
Institut für Mikrobiologie, Immunologie und Hygiene, Technische Universität München, Munich, Germany
Toxoplasma gondii forms different life stages, fast-replicating tachyzoites and slow-growing bradyzoites, in mammalian hosts. CD8 T cells are of crucial importance in toxoplasmosis, but it is unknown which parasite stage is recognized by CD8 T cells. To analyze stage-specific CD8 T cell responses, we generated various recombinant Toxoplasma gondii expressing the heterologous Ag 
-galactosidase (-gal) and studied whether 1) secreted or cytoplasmic Ags and 2) tachyzoites or bradyzoites, which persist intracerebrally, induce CD8 T cells. We monitored the frequencies and kinetics of -gal-specific CD8 T cells in infected mice by MHC class I tetramer staining. Upon oral infection of B6C (H-2bxd) mice, only -gal-secreting tachyzoites induced -gal-specific CD8 T cells. However, upon secondary infection of mice that had received a primary infection with tachyzoites secreting -gal, -gal-secreting tachyzoites and bradyzoites transiently increased the frequency of intracerebral -gal-specific CD8 T cells. Frequencies of splenic and cerebral -gal-specific CD8 T cells peaked at day 23 after infection, thereafter persisting at high levels in the brain but declining in the spleen. Splenic and cerebral -gal-specific CD8 T cells produced IFN-
and were cytolytic upon specific restimulation. Thus, compartmentalization and stage specificity of an Ag determine the induction of CD8 T cells in toxoplasmosis.
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