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Journal of Clinical Microbiology, July 2004, p. 3219-3224, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3219-3224.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Immunohistostaining Assays for Detection of Chlamydia pneumoniae in Atherosclerotic Arteries Indicate Cross-Reactions with Nonchlamydial Plaque Constituents

Vicky Y. Hoymans,1* Johan M. Bosmans,1 Dominique Ursi,2 Wim Martinet,3 Floris L. Wuyts,4 Eric Van Marck,5 Martin Altwegg,6 Christiaan J. Vrints,1 and Margareta M. Ieven2

Departments of Cardiology,1 Microbiology,2 Pharmacology,3 Pathology,5 Medical Statistics, University of Antwerp-UA, Antwerp, Belgium,4 Department of Medical Microbiology, University of Zurich, Zurich, Switzerland6

Received 5 March 2004/ Returned for modification 28 March 2004/ Accepted 11 April 2004

Detection of Chlamydia pneumoniae antigens in PCR-negative atheromata by immunohistochemistry assays has given rise to controversies regarding a link between the bacterium and atherosclerosis. One hundred ninety-seven human arterial segments removed surgically were examined for C. pneumoniae DNA by conventional PCR with three different primer pairs and by real-time PCR in two different laboratories. No C. pneumoniae DNA was detected. Eighty atherosclerotic lesions were studied by immunohistochemistry assays. Immunoreactivity for C. pneumoniae was frequently present but was not related to the extent of atherosclerosis. Mammary arteries showed immunoreactivity. Serial sections of 17 atheromata were analyzed by Western blotting, histological staining, and UV fluorescence microscopy. Chlamydial proteins were not detected. The sites with positive results by C. pneumoniae immunohistostaining assays precisely matched the sites with autofluorescent ceroid deposits. Immunoblotting and antigenic staining for C. pneumoniae were negative in tests with fetal aortas. The absence of C. pneumoniae DNA in human atherosclerotic lesions, together with negative results for C. pneumoniae proteins by Western blotting analysis, and the perfect matching of C. pneumoniae immunoreactive sites with sites with autofluorescent ceroid deposits suggest a nonspecific reactivity of antichlamydial antibodies with plaque constituents. On the basis of the results of the present study, there are no arguments for an etiologic role of C. pneumoniae in atherosclerosis.

* Corresponding author. Mailing address: Division of Cardiology, University of Antwerp (UA), B-2650 Edegem, Belgium. Phone: 32-3-821 49 46. Fax: 32-3-830 23 05. E-mail: vicky.hoymans{at}uza.be.

Journal of Clinical Microbiology, July 2004, p. 3219-3224, Vol. 42, No. 7
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.7.3219-3224.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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