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Infection and Immunity, June 2002, p. 2976-2981, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.2976-2981.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Proteomic Analysis of Differentially Expressed Chlamydia pneumoniae Genes during Persistent Infection of HEp-2 Cells

Robert E. Molestina,1,2 Jon B. Klein,2,3,4,5 Richard D. Miller,6 William H. Pierce,7 Julio A. Ramirez,1,5 and James T. Summersgill1,6*

Division of Infectious Diseases, Department of Medicine,1 Core Proteomics Laboratory,2 Division of Nephrology, Department of Medicine,3 Department of Biochemistry and Molecular Biology,4 Department of Microbiology and Immunology,6 Mass Spectrometry Laboratory, Department of Pharmacology and Toxicology, University of Louisville School of Medicine, University of Louisville, Louisville, Kentucky 40292,7 Veterans Administration Medical Center, Louisville, Kentucky 402065

Received 24 September 2001/ Returned for modification 3 January 2002/ Accepted 14 March 2002

Recent data have shown that the respiratory pathogen Chlamydia pneumoniae expresses an altered gene transcription profile during gamma interferon (IFN-{gamma})-induced persistent infection in vitro. In the present study, we examined, by proteomics, expression of C. pneumoniae proteins labeled intracellularly with [35S]methionine/cysteine under normal conditions or IFN-{gamma}-mediated persistence. The identity of differentially expressed proteins during persistent infection was determined by matching spots to those of proteins identified in C. pneumoniae elementary bodies by matrix-assisted laser desorption ionization mass spectrometry. Upon treatment with 50 U of IFN-{gamma} per ml, a marked upregulation of major outer membrane protein (MOMP), heat shock protein 60 (Hsp-60/GroEL), and proteins with functions in DNA replication (GyrA), transcription (RpoA, PnP), translation (Rrf), glycolysis (PgK, GlgP), and type III secretion (SctN) was observed at 24 h of infection. In contrast, no significant decreases in bacterial protein expression were found in C. pneumoniae-infected cells due to IFN-{gamma} treatment. Upregulation of C. pneumoniae proteins involved in diverse functions during persistent infection may allow the organism to resist the inhibitory effects of IFN-{gamma} while retaining basic functions. Future studies should examine the differential expression of chlamydial proteins during the developmental cycle under IFN-{gamma} pressure to obtain a finer representation of the gene products involved in establishing persistence.

* Corresponding author. Mailing address: Infectious Diseases Laboratory, Instructional Building, Room 311, 500 South Preston St., University of Louisville, Louisville, KY 40292. Phone: (502) 852-5132. Fax: (502) 852-1512. E-mail: jtsumm{at}louisville.edu.

Editor: R. N. Moore

Infection and Immunity, June 2002, p. 2976-2981, Vol. 70, No. 6
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.6.2976-2981.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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