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Infection and Immunity, February 2006, p. 940-946, Vol. 74, No. 2
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.2.940-946.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Activation of Toll-Like Receptor 9 by DNA from Different Bacterial Species

Alexander Dalpke,1,2* Jutta Frank,1,{dagger} Mirjam Peter,2 and Klaus Heeg2

Institute of Medical Microbiology, Philipps-University Marburg, Marburg, Germany,1 Department of Hygiene and Medical Microbiology, University Hospital Heidelberg, Heidelberg, Germany2

Received 2 August 2005/ Returned for modification 4 September 2005/ Accepted 30 October 2005

Toll-like receptor 9 (TLR-9) recognizes unmethylated CpG dinucleotides which are abundant in prokaryotic DNA and yet are rare in eukaryotic DNA. Little is known about the significance of TLR-9 in terms of recognition of different bacterial DNA species. In this study HEK293 cells stably transfected with human TLR-9 were used to analyze the immunostimulatory properties of 15 bacterial DNA preparations. In addition, bacterial genome data were analyzed for the frequency of unmethylated cytosine-guanosine ([CG]) dinucleotides. We observed that DNA samples of different bacteria showed considerable differences in their potential to stimulate TLR-9. This correlated with the frequency of [CG] dinucleotides. Based upon data from our experiments the estimate of immunostimulatory bacterial DNA concentrations translated to as high as 109 bacteria/ml. Application of the transfection reagent DOTAP resulted in a more efficient delivery of DNA into the cell, and this went along with increased TLR-9 activation. The data indicate that bacterial DNA preparations from different species differ in their capacity to activate TLR-9, which is dependent on the individual [CG] content. Moreover, increased intracellular delivery results in a marked enhancement of immunostimulation.

* Corresponding author. Mailing address: Department of Hygiene and Medical Microbiology, Hygiene Institute, University Hospital Heidelberg, Im Neuenheimer Feld 324, D-69120 Heidelberg, Germany. Phone: 49 6221 56 38173. Fax: 49 6221 56 5857. E-mail: alexander.dalpke{at}med.uni-heidelberg.de.

Editor: J. D. Clements

{dagger} Present address: Institute for Medical Parasitology, University Hospital Bonn, Bonn, Germany.

Infection and Immunity, February 2006, p. 940-946, Vol. 74, No. 2
0019-9567/06/$08.00+0     doi:10.1128/IAI.74.2.940-946.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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