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Clinical and Diagnostic Laboratory Immunology, November 1999, p. 819-825, Vol. 6, No. 6
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

Analysis of the Humoral Immune Response to Chlamydia pneumoniae by Immunoblotting and Immunoprecipitation

Andreas Essig,* Ulrike Simnacher, Milorad Susa, and Reinhard Marre

Department of Medical Microbiology and Hygiene, University of Ulm, D-89081 Ulm, Germany

Received 19 April 1999/Returned for modification 23 June 1999/Accepted 26 July 1999

Chlamydia pneumoniae is a widely spread agent of respiratory tract infections in humans. A reliable serodiagnosis of the disease is hampered by the poor knowledge about immunodominant antigens in C. pneumoniae infections. We applied a novel strategy to identify immunogenic proteins of C. pneumoniae TW183 combining metabolic radiolabeling of de novo-synthesized chlamydial antigens with immunoprecipitation. By this technique C. pneumoniae antigens of approximately 160, 97 to 99, 60 to 62, 40, 27, and 15 kDa were detected in the vast majority of sera from patients with a current C. pneumoniae infection. By immunoblotting purified elementary bodies of C. pneumoniae TW183 with the same sera, only the 60- to 62-kDa antigen could be detected consistently. Sequential immunoprecipitation performed at different stages of the chlamydial developmental cycle revealed that the 60- to 62-kDa antigen is strongly upregulated after 24 to 48 h of host cell infection and is presented as a major immunogen in both C. pneumoniae-infected patients and mice. We conclude that, due to its high sensitivity and concurrent preservation of conformational epitopes, metabolic radiolabeling of chlamydial antigens combined with immunoprecipitation may be a useful method to reveal important immunogens in respiratory C. pneumoniae infection which might have been missed by immunoblot analysis.

* Corresponding author. Mailing address: Department of Medical Microbiology and Hygiene, University of Ulm, Robert-Koch Str. 8, D-89081 Ulm, Germany. Phone: 49-731-5024614 or -5024601. Fax: 49-731-5024619. E-mail: andreas.essig{at}medizin.uni-ulm.de.

Clinical and Diagnostic Laboratory Immunology, November 1999, p. 819-825, Vol. 6, No. 6
1071-412X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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