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Applied and Environmental Microbiology, October 2007, p. 6637-6643, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.00923-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Propagation of Arthropod-Borne Rickettsia spp. in Two Mosquito Cell Lines{triangledown}

Joyce M. Sakamoto* and Abdu F. Azad

Department of Microbiology and Immunology, School of Medicine, 660 West Redwood Street, Howard Hall, Room 324B, University of Maryland, Baltimore, Maryland 21201

Received 24 April 2007/ Accepted 16 August 2007

Rickettsiae are obligate intracellular alphaproteobacteria that include pathogenic species in the spotted fever, typhus, and transitional groups. The development of a standardized cell line in which diverse rickettsiae can be grown and compared would be highly advantageous to investigate the differences among and between pathogenic and nonpathogenic species of rickettsiae. Although several rickettsial species have been grown in tick cells, tick cells are more difficult to maintain and they grow more slowly than insect cells. Rickettsia-permissive arthropod cell lines that can be passaged rapidly are highly desirable for studies on arthropod-Rickettsia interactions. We used two cell lines (Aedes albopictus cell line Aa23 and Anopheles gambiae cell line Sua5B) that have not been used previously for the purpose of rickettsial propagation. We optimized the culture conditions to propagate one transitional-group rickettsial species (Rickettsia felis) and two spotted-fever-group rickettsial species (R. montanensis and R. peacockii) in each cell line. Both cell lines allowed the stable propagation of rickettsiae by weekly passaging regimens. Stable infections were confirmed by PCR, restriction digestion of rompA, sequencing, and the direct observation of bacteria by fluorescence in situ hybridization. These cell lines not only supported rickettsial growth but were also permissive toward the most fastidious species of the three, R. peacockii. The permissive nature of these cell lines suggests that they may potentially be used to isolate novel rickettsiae or other intracellular bacteria. Our results have important implications for the in vitro maintenance of uncultured rickettsiae, as well as providing insights into Rickettsia-arthropod interactions.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, School of Medicine, 660 West Redwood St., Howard Hall, Room 324B, University of Maryland, Baltimore, MD 21201. Phone: (410) 706-3337. Fax: (410) 706-0282. E-mail: jsaka001{at}umaryland.edu

{triangledown} Published ahead of print on 31 August 2007.

Applied and Environmental Microbiology, October 2007, p. 6637-6643, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.00923-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Sunyakumthorn, P., Bourchookarn, A., Pornwiroon, W., David, C., Barker, S. A., Macaluso, K. R. (2008). Characterization and Growth of Polymorphic Rickettsia felis in a Tick Cell Line. Appl. Environ. Microbiol. 74: 3151-3158 [Abstract] [Full Text]  


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