Bacterial antigen expression is an important component in inducing an immune response to orally administered Salmonella-delivered DNA vaccines

Michelle E Gahan, Diane E Webster, Steven L Wesselingh, Richard A Strugnell, Ji Yang

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

BACKGROUND: The use of Salmonella to deliver heterologous antigens from DNA vaccines is a well-accepted extension of the success of oral Salmonella vaccines in animal models. Attenuated S. typhimurium and S. typhi strains are safe and efficacious, and their use to deliver DNA vaccines combines the advantages of both vaccine approaches, while complementing the limitations of each technology. An important aspect of the basic biology of the Salmonella/DNA vaccine platform is the relative contributions of prokaryotic and eukaryotic expression in production of the vaccine antigen. Gene expression in DNA vaccines is commonly under the control of the eukaryotic cytomegalovirus (CMV) promoter. The aim of this study was to identify and disable putative bacterial promoters within the CMV promoter and evaluate the immunogenicity of the resulting DNA vaccine delivered orally by S. typhimurium.

METHODOLOGY/PRINCIPAL FINDINGS: The results reported here clearly demonstrate the presence of bacterial promoters within the CMV promoter. These promoters have homology to the bacterial consensus sequence and functional activity. To disable prokaryotic expression from the CMV promoter a series of genetic manipulations were performed to remove the two major bacterial promoters and add a bacteria transcription terminator downstream of the CMV promoter. S. typhimurium was used to immunise BALB/c mice orally with a DNA vaccine encoding the C-fragment of tetanus toxin (TT) under control of the original or the modified CMV promoter. Although both promoters functioned equally well in eukaryotic cells, as indicated by equivalent immune responses following intramuscular delivery, only the original CMV promoter was able to induce an anti-TT specific response following oral delivery by S. typhimurium.

CONCLUSIONS: These findings suggest that prokaryotic expression of the antigen and co-delivery of this protein by Salmonella are at least partially responsible for the successful oral delivery of C-fragment DNA vaccines containing the CMV promoter by S. typhimurium.

Original languageEnglish
Article numbere6062
Pages (from-to)1-9
Number of pages9
JournalPLoS One
Volume4
Issue number6
DOIs
Publication statusPublished - 26 Jun 2009
Externally publishedYes

Fingerprint

bacterial antigens
Bacterial Antigens
DNA Vaccines
Salmonella
recombinant vaccines
Cytomegalovirus
promoter regions
immune response
Salmonella Vaccines
Vaccines
Heterophile Antigens
mouth
Tetanus Toxin
Antigens
tetanus
vaccines
antigens
Transcription
Salmonella Typhimurium
Gene expression

Cite this

Gahan, Michelle E ; Webster, Diane E ; Wesselingh, Steven L ; Strugnell, Richard A ; Yang, Ji. / Bacterial antigen expression is an important component in inducing an immune response to orally administered Salmonella-delivered DNA vaccines. In: PLoS One. 2009 ; Vol. 4, No. 6. pp. 1-9.
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Bacterial antigen expression is an important component in inducing an immune response to orally administered Salmonella-delivered DNA vaccines. / Gahan, Michelle E; Webster, Diane E; Wesselingh, Steven L; Strugnell, Richard A; Yang, Ji.

In: PLoS One, Vol. 4, No. 6, e6062, 26.06.2009, p. 1-9.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Bacterial antigen expression is an important component in inducing an immune response to orally administered Salmonella-delivered DNA vaccines

AU - Gahan, Michelle E

AU - Webster, Diane E

AU - Wesselingh, Steven L

AU - Strugnell, Richard A

AU - Yang, Ji

PY - 2009/6/26

Y1 - 2009/6/26

N2 - BACKGROUND: The use of Salmonella to deliver heterologous antigens from DNA vaccines is a well-accepted extension of the success of oral Salmonella vaccines in animal models. Attenuated S. typhimurium and S. typhi strains are safe and efficacious, and their use to deliver DNA vaccines combines the advantages of both vaccine approaches, while complementing the limitations of each technology. An important aspect of the basic biology of the Salmonella/DNA vaccine platform is the relative contributions of prokaryotic and eukaryotic expression in production of the vaccine antigen. Gene expression in DNA vaccines is commonly under the control of the eukaryotic cytomegalovirus (CMV) promoter. The aim of this study was to identify and disable putative bacterial promoters within the CMV promoter and evaluate the immunogenicity of the resulting DNA vaccine delivered orally by S. typhimurium.METHODOLOGY/PRINCIPAL FINDINGS: The results reported here clearly demonstrate the presence of bacterial promoters within the CMV promoter. These promoters have homology to the bacterial consensus sequence and functional activity. To disable prokaryotic expression from the CMV promoter a series of genetic manipulations were performed to remove the two major bacterial promoters and add a bacteria transcription terminator downstream of the CMV promoter. S. typhimurium was used to immunise BALB/c mice orally with a DNA vaccine encoding the C-fragment of tetanus toxin (TT) under control of the original or the modified CMV promoter. Although both promoters functioned equally well in eukaryotic cells, as indicated by equivalent immune responses following intramuscular delivery, only the original CMV promoter was able to induce an anti-TT specific response following oral delivery by S. typhimurium.CONCLUSIONS: These findings suggest that prokaryotic expression of the antigen and co-delivery of this protein by Salmonella are at least partially responsible for the successful oral delivery of C-fragment DNA vaccines containing the CMV promoter by S. typhimurium.

AB - BACKGROUND: The use of Salmonella to deliver heterologous antigens from DNA vaccines is a well-accepted extension of the success of oral Salmonella vaccines in animal models. Attenuated S. typhimurium and S. typhi strains are safe and efficacious, and their use to deliver DNA vaccines combines the advantages of both vaccine approaches, while complementing the limitations of each technology. An important aspect of the basic biology of the Salmonella/DNA vaccine platform is the relative contributions of prokaryotic and eukaryotic expression in production of the vaccine antigen. Gene expression in DNA vaccines is commonly under the control of the eukaryotic cytomegalovirus (CMV) promoter. The aim of this study was to identify and disable putative bacterial promoters within the CMV promoter and evaluate the immunogenicity of the resulting DNA vaccine delivered orally by S. typhimurium.METHODOLOGY/PRINCIPAL FINDINGS: The results reported here clearly demonstrate the presence of bacterial promoters within the CMV promoter. These promoters have homology to the bacterial consensus sequence and functional activity. To disable prokaryotic expression from the CMV promoter a series of genetic manipulations were performed to remove the two major bacterial promoters and add a bacteria transcription terminator downstream of the CMV promoter. S. typhimurium was used to immunise BALB/c mice orally with a DNA vaccine encoding the C-fragment of tetanus toxin (TT) under control of the original or the modified CMV promoter. Although both promoters functioned equally well in eukaryotic cells, as indicated by equivalent immune responses following intramuscular delivery, only the original CMV promoter was able to induce an anti-TT specific response following oral delivery by S. typhimurium.CONCLUSIONS: These findings suggest that prokaryotic expression of the antigen and co-delivery of this protein by Salmonella are at least partially responsible for the successful oral delivery of C-fragment DNA vaccines containing the CMV promoter by S. typhimurium.

KW - Administration, Oral

KW - Animals

KW - Antibodies, Bacterial

KW - Antigens, Bacterial

KW - Base Sequence

KW - Cytomegalovirus

KW - Female

KW - Immune System

KW - Mice

KW - Mice, Inbred BALB C

KW - Molecular Sequence Data

KW - Promoter Regions, Genetic

KW - Salmonella Vaccines

KW - Vaccines, DNA

KW - beta-Galactosidase

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

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DO - 10.1371/journal.pone.0006062

M3 - Article

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EP - 9

JO - PLoS One

JF - PLoS One

SN - 1932-6203

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M1 - e6062

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