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Infection and Immunity, January 2005, p. 362-368, Vol. 73, No. 1
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.1.362-368.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Optimization of Salmonella enterica Serovar Typhi {Delta}aroC {Delta}ssaV Derivatives as Vehicles for Delivering Heterologous Antigens by Chromosomal Integration and In Vivo Inducible Promoters

Richard Stratford,1* Nicola D. McKelvie,1 Nicky J. Hughes,1 Emma Aldred,1 Claire Wiseman,1 Johanna Curtis,1 Trevor Bellaby,1 Matt Bentley,1 Zoë Hindle,1 Frank R. Brennan,1 Steven N. Chatfield,1 Gordon Dougan,2 and Shahid A. Khan1

Microscience Limited, Winnersh Triangle, Wokingham, Berkshire,1 Centre for Molecular Microbiology and Infection, Department of Biological Sciences, Imperial College of Science, Technology and Medicine, London, United Kingdom2

Received 5 May 2004/ Returned for modification 9 June 2004/ Accepted 2 August 2004

Novel candidate live oral vaccines based on a Salmonella enterica serovar Typhi ZH9 (Ty2 {Delta}aroC {Delta}ssaV) derivative that directed the expression of either the B subunit of Escherichia coli heat-labile toxin or hepatitis B virus core antigen from the bacterial chromosome using the in vivo inducible ssaG promoter were constructed. The levels of attenuation of the two S. enterica serovar Typhi ZH9 derivatives were similar to that of the parent as assessed by measuring the replication of bacteria within human macrophage-like U937 cells. The expression of heterologous antigen in the respective S. enterica serovar Typhi ZH9 derivatives was up-regulated significantly within U937 cells compared to similar S. enterica serovar Typhi ZH9 derivative bacteria grown in modified Luria-Bertani broth supplemented with aromatic amino acids. Immunization of mice with these S. enterica serovar Typhi ZH9 derivatives stimulated potent antigen-specific serum immunoglobulin G responses to the heterologous antigens.

* Corresponding author. Mailing address: Microscience Ltd., 540-545 Eskdale Rd., Winnersh Triangle, Wokingham, Berkshire RG41 5TU, United Kingdom. Phone: 44 (0) 118 944 3356. Fax: 44 (0) 118 944 3301. E-mail: r.stratford{at}microscience.com.

Editor: J. D. Clements

Infection and Immunity, January 2005, p. 362-368, Vol. 73, No. 1
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.1.362-368.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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