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Immunoproteomic Discovery of Novel T Cell Antigens from the Obligate Intracellular Pathogen Chlamydia¹

Karuna P. Karunakaran^*, Jose Rey-Ladino^*, Nikolay Stoynov, Kyra Berg^*, Caixia Shen^*, Xiaozhou Jiang^*, Brent R. Gabel^*, Hong Yu^*, Leonard J. Foster^2, and Robert C. Brunham^2,*

^* British Columbia Centre for Disease Control and Department of Biochemistry and Molecular Biology, University of British Columbia Centre for Proteomics, University of British Columbia, Vancouver, British Columbia, Canada

Chlamydia infections cause substantial morbidity worldwide and effective prevention will depend on a vaccine. Since Chlamydia immunity is T cell-mediated, a major impediment to developing a molecular vaccine has been the difficulty in identifying relevant T cell Ags. In this study, we used a combination of affinity chromatography and tandem mass spectrometry to identify 13 Chlamydia peptides among 331 self-peptides presented by MHC class II (I-A^b) molecules from bone marrow-derived murine dendritic cells infected with Chlamydia muridarum. These MHC class II-bound peptides were recognized by Chlamydia-specific CD4 T cells harvested from immune mice and adoptive transfer of dendritic cells pulsed ex vivo with the peptides partially protected mice against intranasal and genital tract Chlamydia infection. The results provide evidence for lead vaccine candidates for a T cell-based subunit molecular vaccine against Chlamydia infection suitable for human study.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ This work was supported by Canadian Institutes of Health Research and Genome British Columbia grants (to R.C.B.) and Canadian Institutes of Health Operating Grant MOP-77688 (to L.J.F.). L.J.F. is a Michael Smith Foundation Scholar and the Canada Research Chair in Organelle Proteomics.

² Address correspondence and reprint requests to Dr. Robert C. Brunham, British Columbia Centre for Disease Control, 655 West 12th Avenue, Vancouver, British Columbia V5Z 4R4, Canada. E-mail address: robert.brunham{at}bccdc.ca or Dr. Leonard J. Foster, Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada. E-mail address: ljfoster{at}interchange.ubc.ca

³ Abbreviations used in this paper: CMI, cell-mediated immune response; DC, dendritic cell; BMDC, bone marrow-derived DC; MS/MS, tandem mass spectrometry; IFU, inclusion-forming units; EB, elementary body; HK-EB, heat-killed EB.

⁴ The online version of this article contains supplemental material.