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Infection and Immunity, July 2006, p. 4254-4265, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00386-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

cDNA Microarray Analysis of Host-Pathogen Interactions in a Porcine In Vitro Model for Toxoplasma gondii Infection{dagger}

Margaret Okomo-Adhiambo,1* Craig Beattie,1 and Anette Rink1,2

Department of Animal Biotechnology, University of Nevada, 1664 North Virginia Street, Reno, Nevada 89557,1 Animal Disease and Food Safety Laboratory, Nevada Department of Agriculture, 350 Capitol Hill Avenue, Reno, Nevada 895022

Received 9 March 2005/ Returned for modification 19 April 2005/ Accepted 14 April 2006

Toxoplasma gondii induces the expression of proinflammatory cytokines, reorganizes organelles, scavenges nutrients, and inhibits apoptosis in infected host cells. We used a cDNA microarray of 420 annotated porcine expressed sequence tags to analyze the molecular basis of these changes at eight time points over a 72-hour period in porcine kidney epithelial (PK13) cells infected with T. gondii. A total of 401 genes with Cy3 and Cy5 spot intensities of ≥500 were selected for analysis, of which 263 (65.6%) were induced ≥2-fold (expression ratio, ≥2.0; P ≤ 0.05 [t test]) over at least one time point and 48 (12%) were significantly down-regulated. At least 12 functional categories of genes were modulated (up- or down-regulated) by T. gondii. The majority of induced genes were clustered as transcription, signal transduction, host immune response, nutrient metabolism, and apoptosis related. The expression of selected genes altered by T. gondii was validated by quantitative real-time reverse transcription-PCR. These results suggest that significant changes in gene expression occur in response to T. gondii infection in PK13 cells, facilitating further analysis of host-pathogen interactions in toxoplasmosis in a secondary host.

* Corresponding author. Mailing address: Department of Animal Biotechnology, University of Nevada, 1664 North Virginia Street, Reno, NV 89557. Phone: (775) 784-6135. Fax: (775) 784-1375. E-mail: mokomo{at}cabnr.unr.edu.

{dagger} Supplemental material for this article may be found at http://iai.asm.org/.

Editor: J. F. Urban, Jr.

Infection and Immunity, July 2006, p. 4254-4265, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00386-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.





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