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Immunodominant, protective response to the parasite Toxoplasma gondii requires antigen processing in the endoplasmic reticulum

Nature Immunology volume 9, pages 937–944 (2008)Cite this article

Abstract

The parasite Toxoplasma gondii replicates in a specialized intracellular vacuole and causes disease in many species. Protection from toxoplasmosis is mediated by CD8+ T cells, but the T. gondii antigens and host genes required for eliciting protective immunity are poorly defined. Here we identified GRA6, a polymorphic protein secreted in the parasitophorous vacuole, as the source of the immunodominant and protective decapeptide HF10 presented by the H-2Ld major histocompatibility complex class I molecule. Presentation of the HF10–H-2Ld ligand required proteolysis by ERAAP, the endoplasmic reticulum aminopeptidase associated with antigen processing. Consequently, expansion of protective CD8+ T cell populations was impaired in T. gondii–infected ERAAP-deficient mice, which were more susceptible to toxoplasmosis. Thus, endoplasmic reticulum proteolysis is critical for eliciting protective immunity to a vacuolar parasite.

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Figure 1: Uncontrolled parasite replication in ERAAP-deficient H-2d mice and greater susceptibility to T. gondii infection.
Figure 2: All T. gondii–specific CD8+ T cell hybridomas are stimulated by H-2Ld MHC class I.
Figure 3: T. gondii–specific CD8+ T cell hybrids recognize the HF10 decapeptide presented by H-2Ld MHC class I.
Figure 4: The HF10–H-2Ld complex is the only detectable ligand recognized by CD8+ T cells in T. gondii–infected H-2d mice.
Figure 5: Immunization with HF10 protects mice from toxoplasmosis.
Figure 6: Presentation of the HF10–H-2Ld complex requires proteasomal activity and TAP transport.
Figure 7: Presentation of HF10–H-2Ld complexes requires proteolysis in the endoplasmic reticulum.

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Acknowledgements

We thank J. Boothroyd (Stanford University) for the parental Pru strain of T. gondii and the GFP-expressing derivative; R. McLeod (University of Chicago) for B1 internal control DNA; F. Levy (University of Epalinges) for the pEGFP-Ub vector; R. Vance (University of California, Berkeley) for the colony-stimulating factor–producing 3T3 cells; W. Sha (University of California, Berkeley) for the pMSCV2.2 retroviral vector; D. King (University of California, Berkeley) for peptide synthesis; J. Boothroyd and R. McLeod for discussions; the Center for Host-Pathogen Studies core facilities and S.-W. Chan for technical assistance; and B. Striepen for comments on the manuscript. Supported by the US National Institutes of Health (N.S. and E.A.R.) and the international Human Frontier Science Program (LT00841/2005-l to N.B.).

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  1. Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, Berkeley, California 94720, USA.,

    Nicolas Blanchard, Federico Gonzalez, Marie Schaeffer, Nathalie T Joncker, Tiffany Cheng, Anjali J Shastri, Ellen A Robey & Nilabh Shastri

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Contributions

N.B. designed and did all experiments in collaboration with F.G. (cDNA library), M.S. (brain cell isolation), N.T.J. (footpad immunization and measurement of the natural killer cell response), T.C. (processed peptide analysis) and A.J.S. (parasite burden measurements); E.A.R. and N.S. provided guidance and contributed to the experimental design; and N.B. and N.S. wrote the manuscript together with input from other authors.

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Correspondence to Nicolas Blanchard or Nilabh Shastri.

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Blanchard, N., Gonzalez, F., Schaeffer, M. et al. Immunodominant, protective response to the parasite Toxoplasma gondii requires antigen processing in the endoplasmic reticulum. Nat Immunol 9, 937–944 (2008). https://doi.org/10.1038/ni.1629

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