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The Journal of Immunology, 2008, 180: 5490-5498.
Copyright © 2008 by The American Association of Immunologists, Inc.

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*Chlamydia Infections

Immunoproteomic Identification and Serological Responses to Novel Chlamydia pneumoniae Antigens That Are Associated with Persistent C. pneumoniae Infections1

Sebastian Bunk2,*, Iuliana Susnea2,{dagger}, Jan Rupp{ddagger}, James T. Summersgill§, Matthias Maass, Werner Stegmann||, André Schrattenholz||, Albrecht Wendel*, Michael Przybylski{dagger} and Corinna Hermann3,*

* Department of Biochemical Pharmacology and {dagger} Department of Analytical Chemistry, University of Konstanz, Konstanz, Germany; {ddagger} Institute of Medical Microbiology and Hygiene, University of Luebeck, Luebeck, Germany; § Division of Infectious Diseases, Department of Medicine, University of Louisville, Louisville, KY 40292; Institute of Medical Microbiology, Hygiene, and Infectious Diseases, Paracelsus Private Medical University and University Hospital Salzburg, Salzburg, Austria; and || ProteoSys, Mainz, Germany

The controversial discussion about the role of Chlamydia pneumoniae in atherosclerosis cannot be solved without a reliable diagnosis that allows discrimination between past and persistent infections. Using a proteomic approach and immunoblotting with human sera, we identified 31 major C. pneumoniae Ags originating from 27 different C. pneumoniae proteins. More than half of the proteins represent Chlamydia Ags not described previously. Using a comparative analysis of spot reactivity Pmp6, OMP2, GroEL, DnaK, RpoA, EF-Tu, as well as CpB0704 and CpB0837, were found to be immunodominant. The comparison of Ab-response patterns of sera from subjects with and without evidence for persisting C. pneumoniae, determined by multiple PCR analysis of PBMC and vasculatory samples, resulted in differential reactivity for 12 proteins, which is not reflected by reactivity of the sera in the microimmunofluorescence test, the current gold standard for serodiagnosis. Although reactivity of sera from PCR-positive donors was increased toward RpoA, MOMP, YscC, Pmp10, PorB, Pmp21, GroEL, and Cpaf, the reactivity toward YscL, Rho, LCrE, and CpB0837 was decreased, reflecting the altered protein expression of persisting C. pneumoniae in vitro. Our data provide the first evidence of a unique Ab-response pattern associated with persistent C. pneumoniae infections, which is a prerequisite for the serological determination of persistently infected patients.

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 the Eliteprogramm für Postdoktorandinnen und Postdoktoranden der Landesstiftung Baden-Württemberg, the German Research Council FOR 434, and, in part, by the Austrian Federal Ministry of Science and Research/GEN-Au within the European Research Area-Network PathoGenoMics: European Initiative to Fight Chlamydial Infections by Unbiased Genomics, and German Research Council Grant SPP1130: Ma2070-4/3.

2 S.B. and I.S. contributed equally to this publication.

3 Address correspondence and reprint requests to Dr. Corinna Hermann, Department of Biochemical Pharmacology, University of Konstanz, P.O. Box M668, 78457 Konstanz, Germany. E-mail address: Corinna.hermann{at}uni-konstanz.de

4 Abbreviations used in this paper: MIF, microimmunofluorescence; EB, elementary body; 2-D, two dimensional; IPG, immobilized pH gradient; FT-ICR, Fourier transform ion cyclotron resonance; RT, room temperature; pI, isoelectric point; Pmp, polymorphic membrane protein.







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