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Infection and Immunity, April 2003, p. 1944-1952, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.1944-1952.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Autodisplay: Development of an Efficacious System for Surface Display of Antigenic Determinants in Salmonella Vaccine Strains

Uwe Kramer,1 Konstantin Rizos,2,{dagger} Heiko Apfel,2 Ingo B. Autenrieth,1 and Claus T. Lattemann2*

Institut für Medizinische Mikrobiologie und Krankenhaushygiene der Universität Tübingen, D-72076 Tübingen,1 CREATOGEN AG, D-86156 Augsburg, Germany2

Received 5 July 2002/ Returned for modification 30 September 2002/ Accepted 18 December 2002

To optimize antigen delivery by Salmonella vaccine strains, a system for surface display of antigenic determinants was established by using the autotransporter secretion pathway of gram-negative bacteria. A modular system for surface display allowed effective targeting of heterologous antigens or fragments thereof to the bacterial surface by the autotransporter domain of AIDA-I, the Escherichia coli adhesin involved in diffuse adherence. A major histocompatibility complex class II-restricted epitope, comprising amino acids 74 to 86 of the Yersinia enterocolitica heat shock protein Hsp60 (Hsp6074-86), was fused to the AIDA-I autotransporter domain, and the resulting fusion protein was expressed at high levels on the cell surface of E. coli and Salmonella enterica serovar Typhimurium. Colonization studies in mice vaccinated with Salmonella strains expressing AIDA-I fusion proteins demonstrated high genetic stability of the generated vaccine strain in vivo. Furthermore, a pronounced T-cell response against Yersinia Hsp6074-86 was induced in mice vaccinated with a Salmonella vaccine strain expressing the Hsp6074-86-AIDA-I fusion protein. This was shown by monitoring Yersinia Hsp60-stimulated IFN-{gamma} secretion and proliferation of splenic T cells isolated from vaccinated mice. These results demonstrate that the surface display of antigenic determinants by the autotransporter pathway deserves special attention regarding the application in live attenuated Salmonella vaccine strains.

* Corresponding author. Mailing address: Aventis Pharma Deutschland GmbH, Biocenter H780, Industriepark Höchst, 65926 Frankfurt am Main, Germany. Phone: 49-69-30-52-47-95. Fax: 49-69-30-51-63-71. E-mail: claus.lattemann{at}aventis.com.

Editor: A. D. O'Brien

{dagger} Present address: Max Planck-Institut für Infektionsbiologie, Abteilung Molekulare Biologie, 10117 Berlin, Germany.

Infection and Immunity, April 2003, p. 1944-1952, Vol. 71, No. 4
0019-9567/03/$08.00+0     DOI: 10.1128/IAI.71.4.1944-1952.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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