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Infection and Immunity, October 2002, p. 5827-5834, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5827-5834.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Isolation of Chlamydia pneumoniae Clonal Variants by a Focus-Forming Assay

Jens Gieffers,1,2 Robert J. Belland,3 William Whitmire,1 Scot Ouellette,4 Deborah Crane,1 Matthias Maass,2 Gerald I. Byrne,4 and Harlan D. Caldwell1*

Laboratory of Intracellular Parasites,1 Laboratory of Human Bacterial Pathogenesis, National Institutes of AllergyInfectious Disease, National Institutes of Health, Hamilton, Montana 59840,3 Department of Medical Microbiology and Immunology, University of Wisconsin School of Medicine, Madison, Wisconsin 53711,4 Institute of Medical Microbiology and Hygiene, Medical University of Luebeck, Luebeck, Germany2

Received 17 April 2002/ Returned for modification 7 June 2002/ Accepted 18 June 2002

Chlamydia pneumoniae is an obligate intracellular prokaryotic human pathogen that causes community-acquired respiratory infection and has been associated with atherosclerosis and cardiovascular disease. Unexpected results from genomic sequencing indicate that significant intrastrain polymorphism exists for some C. pneumoniae isolates. These polymorphisms could reflect genotypes with differing disease-causing characteristics. A definitive means to test this hypothesis is to obtain genetically homogeneous clonal populations of the pathogen and test them in models of infection and disease. To date, methods for cloning C. pneumoniae have not been reported. In this study, we describe the isolation of clonal variants with genetic differences in the tyrP locus from a polymorphic respiratory isolate, using a novel focus-forming assay. These results now allow investigations on the biology and pathogenesis of C. pneumoniae clonal genovars that could lead to new insights into the pathogenesis of this important human pathogen.

* Corresponding author. Mailing address: Laboratory of Intracellular Parasites, Rocky Mountain Laboratories, NIAID, NIH, 903 South 4th St., Hamilton, MT 59840. Phone: (406) 363-9333. Fax: (406) 363-9355. E-mail: hcaldwell{at}niaid.nih.gov.

Editor: A. D. O'Brien

Infection and Immunity, October 2002, p. 5827-5834, Vol. 70, No. 10
0019-9567/02/$04.00+0     DOI: 10.1128/IAI.70.10.5827-5834.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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