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Infection and Immunity, July 2006, p. 3946-3957, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00112-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Comparison of Different Live Vaccine Strategies In Vivo for Delivery of Protein Antigen or Antigen-Encoding DNA and mRNA by Virulence-Attenuated Listeria monocytogenes

Daniela I. M. Loeffler, Christoph U. Schoen,{dagger} Werner Goebel, and Sabine Pilgrim*

Lehrstuhl für Mikrobiologie der Universität Würzburg, Theodor-Boveri-Institut für Biowissenschaften, Am Hubland, 97074 Würzburg, Germany

Received 22 January 2006/ Returned for modification 20 February 2006/ Accepted 30 March 2006

Listeria monocytogenes can be used to deliver protein antigens or DNA and mRNA encoding such antigens directly into the cytosol of host cells because of its intracellular lifestyle. In this study, we compare the in vivo efficiencies of activation of antigen-specific CD8 and CD4 T cells when the antigen is secreted by L. monocytogenes or when antigen-encoding plasmid DNA or mRNA is released by self-destructing strains of L. monocytogenes. Infection of mice with self-destructing L. monocytogenes carriers delivering mRNA that encodes a nonsecreted form of ovalbumin (OVA) resulted in a significant OVA-specific CD8 T-cell response. In contrast, infection with L. monocytogenes delivering OVA-encoding DNA failed to generate specific T cells. Secretion of OVA by the carrier bacteria yielded the strongest immune response involving OVA-specific CD8 and CD4 T cells. In addition, we investigated the antigen delivery capacity of a self-destructing, virulence-attenuated L. monocytogenes aroA/B mutant. In contrast to the wild-type strain, this mutant exhibited only marginal liver toxicity when high doses (5 x 107 CFU per animal administered intravenously) were used, and it was also able to deliver sufficient amounts of secreted OVA into mice. Therefore, the results presented here could lay the groundwork for a rational combination of L. monocytogenes as an attenuated carrier for the delivery of protein and nucleic acid vaccines in novel vaccination strategies.

* Corresponding author. Present address: Dade Behring Marburg GmbH, P.O. Box 1149, 35001 Marburg, Germany. Phone: (49) 6421 394145. Fax: (49) 6421 394680. E-mail: Sabine_Pilgrim{at}dadebehring.com.

Editor: D. L. Burns

{dagger} Present address: Institut für Hygiene und Mikrobiologie, Josef-Schneider-Str. 2, 97074 Würzburg, Germany.

Infection and Immunity, July 2006, p. 3946-3957, Vol. 74, No. 7
0019-9567/06/$08.00+0     doi:10.1128/IAI.00112-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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