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T-Cell Expression Cloning of Porphyromonas gingivalis Genes Coding for T Helper-Biased Immune Responses during Infection

The Forsyth Institute, Boston, Massachusetts,¹ University of Campinas Piracicaba School of Dentistry, Piracicaba, São Paulo, Brazil,² Infectious Disease Research Institute, Seattle, Washington,³ Deeley Research Centre, BC Cancer Agency, Victoria, British Columbia, Canada,⁴ Henry M. Goldman School of Dental Medicine, Boston University, Boston, Massachusetts⁵

Received 16 December 2005/ Returned for modification 9 February 2006/ Accepted 7 April 2006

Exposure of the mouse oral cavity to Porphyromonas gingivalis results in the development of gingivitis and periapical bone loss, which apparently are associated with a Th1 response to bacterial antigens. We have used this infection model in conjunction with direct T-cell expression cloning to identify bacterial antigens that induce a preferential or biased T helper response during the infectious process. A P. gingivalis-specific CD4 T-cell line derived from mice at 3 weeks postchallenge was used to directly screen a P. gingivalis genomic expression library. This screen resulted in the identification of five genes coding for previously identified proteins and three other putative protein antigens. One of the identified proteins, P. gingivalis thiol peroxidase, was studied in detail because this molecule belongs to a protein family that is apparently involved in microbial pathogenesis. Infection of mice with P. gingivalis, either via the subcutaneous route or after exposure of the animal's oral cavity to viable bacteria, resulted in the induction of a strong thiol peroxidase-specific immune response characterized by the production of high titers of specific serum immunoglobulin G2a antibody and the production of gamma interferon by antigen-stimulated lymphoid cells, a typical Th1-biased response. Thus, the use of a proven T-cell expression cloning approach and a mouse model of periodontal disease resulted in the identification and characterization of P. gingivalis proteins that might be involved in pathogenesis.

Editor: J. F. Urban, Jr.

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