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Applied and Environmental Microbiology, May 2007, p. 3123-3136, Vol. 73, No. 10
0099-2240/07/$08.00+0     doi:10.1128/AEM.01399-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Characterization of Genetically Matched Isolates of Campylobacter jejuni Reveals that Mutations in Genes Involved in Flagellar Biosynthesis Alter the Organism's Virulence Potential

Preeti Malik-Kale,¹ Brian H. Raphael,^1, Craig T. Parker,² Lynn A. Joens,³ John D. Klena,^1, Beatriz Quiñones,² Amy M. Keech,¹ and Michael E. Konkel^1*

School of Molecular Biosciences, Washington State University, Pullman, Washington,¹ USDA, Western Regional Research Center, Albany, California,² Department of Veterinary Science and Microbiology, University of Arizona, Tucson, Arizona³

Received 16 June 2006/ Accepted 28 February 2007

Phenotypic and genotypic evidence suggests that not all Campylobacter jejuni isolates are pathogenic for humans. We hypothesized that differences in gene content or gene expression alter the degree of pathogenicity of C. jejuni isolates. A C. jejuni isolate (Turkey) recovered from a turkey and a second C. jejuni isolate (CS) recovered from a chicken differed in their degrees of in vitro and in vivo virulence. The C. jejuni Turkey isolate invaded INT 407 human epithelial cells and secreted the Cia (Campylobacter invasion antigen) proteins, while the C. jejuni CS isolate was noninvasive for human epithelial cells and did not secrete the Cia proteins. Newborn piglets inoculated with the C. jejuni Turkey isolate developed more severe clinical signs of campylobacteriosis than piglets inoculated with the C. jejuni CS isolate. Additional work revealed that flagellin was not expressed in the C. jejuni CS isolate. Microarray and real-time reverse transcription-PCR analyses revealed that all flagellar class II genes were significantly downregulated in the C. jejuni CS isolate compared to the C. jejuni Turkey isolate. Finally, nucleotide sequencing of the flgR gene revealed the presence of a single residue that was different in the FlgR proteins of the C. jejuni Turkey and CS isolates. Complementation of the C. jejuni CS isolate with a wild-type copy of the flgR gene restored the isolate's motility. Collectively, these findings support the hypothesis that critical differences in gene content or gene expression can alter the pathogenic potential of C. jejuni isolates.

Published ahead of print on 16 March 2007.

Present address: Botulism Public Health Research and Preparedness Laboratory, Centers for Disease Control and Prevention, Atlanta, GA.

Present address: U.S. Naval Medical Research Unit 3, Cairo, Egypt.

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