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Infection and Immunity, October 2001, p. 6172-6178, Vol. 69, No. 10
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.10.6172-6178.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Transcriptional Analysis of p30 Major Outer Membrane Multigene Family of Ehrlichia canis in Dogs, Ticks, and Cell Culture at Different Temperatures

Ahmet Unver,1 Norio Ohashi,1 Tomoko Tajima,1 Roger W. Stich,2 Debra Grover,2 and Yasuko Rikihisa1,*

Department of Veterinary Biosciences1 and Department of Veterinary Preventive Medicine,2 College of Veterinary Medicine, The Ohio State University, Columbus, Ohio 43210-1093

Received 11 April 2001/Returned for modification 11 June 2001/Accepted 9 July 2001

Ehrlichia canis, an obligatory intracellular bacterium of monocytes and macrophages, causes canine monocytic ehrlichiosis. E. canis immunodominant 30-kDa major outer membrane proteins are encoded by a polymorphic multigene family consisting of more than 20 paralogs. In the present study, we analyzed the mRNA expression of 14 paralogs in experimentally infected dogs and Rhipicephalus sanguineus ticks by reverse transcription-PCR using gene-specific primers followed by Southern blotting. Eleven out of 14 paralogs in E. canis were transcribed in increasing numbers and transcription levels, while the mRNA expression of the 3 remaining paralogs was not detected in blood monocytes of infected dogs during the 56-day postinoculation period. Three different groups of R. sanguineus ticks (adult males and females and nymphs) were separately infected with E. canis by feeding on the infected dogs. In these pools of acquisition-fed ticks as well as in the transmission-fed adult ticks, the transcript from only one paralog was detected, suggesting the predominant transcription of that paralog or the suppression of the remaining paralogs in ticks. Expression of the same paralog was higher whereas expression of the remaining paralogs was lower in E. canis cultivated in dog monocyte cell line DH82 at 25°C than in E. canis cultivated at 37°C. Analysis of differential expression of p30 multigenes in dogs, ticks, or monocyte cell cultures would help in understanding the role of these gene products in pathogenesis and E. canis transmission as well as in designing a rational vaccine candidate immunogenic against canine ehrlichiosis.

* Corresponding author. Mailing address: Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, 1925 Coffey Rd., Columbus, OH 43210-1093. Phone: (614) 292-9677. Fax: (614) 292-6473. E-mail: rikihisa.1{at}osu.edu.

Infection and Immunity, October 2001, p. 6172-6178, Vol. 69, No. 10
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.10.6172-6178.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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