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Infection and Immunity, December 2004, p. 7240-7246, Vol. 72, No. 12
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.12.7240-7246.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Development of a System To Study CD4+-T-Cell Responses to Transgenic Ovalbumin-Expressing Toxoplasma gondii during Toxoplasmosis

Marion Pepper,1,{dagger} Florence Dzierszinski,{dagger} Amy Crawford,2 Christopher A. Hunter,1 and David Roos2*

Department of Pathobiology, School of Veterinary Medicine,1 Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania2

Received 19 May 2004/ Returned for modification 23 June 2004/ Accepted 25 August 2004

The study of the immune response to Toxoplasma gondii has provided numerous insights into the role of T cells in resistance to intracellular infections. However, the complexity of this eukaryote pathogen has made it difficult to characterize immunodominant epitopes that would allow the identification of T cells with a known specificity for parasite antigens. As a consequence, analysis of T-cell responses to T. gondii has been based on characterization of the percentage of T cells that express an activated phenotype during infection and on the ability of these cells to produce cytokines in response to complex mixtures of parasite antigens. In order to study specific CD4+ T cells responses to T. gondii, recombinant parasites that express a truncated ovalbumin (OVA) protein, in either a cytosolic or a secreted form, were engineered. In vitro and in vivo studies reveal that transgenic parasites expressing secreted OVA are able to stimulate T-cell receptor-transgenic OVA-specific CD4+ T cells to proliferate, express an activated phenotype, and produce gamma interferon (IFN-{gamma}). Furthermore, the adoptive transfer of OVA-specific T cells into IFN-{gamma}–/– mice provided enhanced protection against infection with the OVA-transgenic (but not parental) parasites. Together, these studies establish the utility of this transgenic system to study CD4+-T-cell responses during toxoplasmosis.

* Corresponding author. Mailing address: Department of Biology, University of Pennsylvania, Philadelphia, PA 19104. Phone: (215) 898-2118. Fax: (215) 746-6697. E-mail: droos{at}sas.upenn.edu.

Editor: J. F. Urban, Jr.

{dagger} M.P. and F.D. contributed equally to this paper.

Infection and Immunity, December 2004, p. 7240-7246, Vol. 72, No. 12
0019-9567/04/$08.00+0     DOI: 10.1128/IAI.72.12.7240-7246.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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