Adaptation of the Endogenous
 Salmonella enterica
 Serovar Typhi
 clyA
 -Encoded Hemolysin for Antigen Export Enhances the Immunogenicity of Anthrax Protective Antigen Domain 4 Expressed by the Attenuated Live-Vector Vaccine Strain CVD 908-
 htrA

Galen, James E.; Zhao, Long; Chinchilla, Magaly; Wang, Jin Yuan; Pasetti, Marcela F.; Green, Jeffrey; Levine, Myron M.

doi:10.1128/iai.72.12.7096-7106.2004

Cited by 65 publications

(67 citation statements)

References 56 publications

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“…In a recent demonstration of the remarkable tunability of OMVs by our group, heterologous proteins fused with the native bacterial protein ClyA are efficiently transported in their native functional forms to vesicles (22). Previous work also points to the utility of ClyA in enhancing the immunogenicity of antigens secreted from a live attenuated Salmonella vector, further suggesting the possibility that antigens may be exported from live vectors in the form of OMVs (23).…”

Section: Discussionmentioning

confidence: 95%

“…1, depicting antibody titers in mice immunized with the purified proteins, show ClyA to be an effective carrier protein for enhancing the immunogenicity of GFP. The immunogenicity of ClyA fusion proteins was first shown in a study by Galen et al (23), where the observed enhancement in the immune response to the antigens was postulated to be largely dependent on export from a live Salmonella vector. Here, the results show that antigen fusion with ClyA is directly responsible for enhanced immunogenicity.…”

Section: Discussionmentioning

confidence: 96%

“…Recent work by our group showed that genetic fusion with ClyA results in the efficient translocation of heterologous proteins to OMVs (22). Additionally, Galen et al (23) showed that antigens fused to ClyA exhibit immunostimulatory activity when secreted from a live Salmonella vaccine.…”

mentioning

confidence: 99%

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Delivery of foreign antigens by engineered outer membrane vesicle vaccines

Chen

Osterrieder

Metzger

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

256

247

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As new disease threats arise and existing pathogens grow resistant to conventional interventions, attention increasingly focuses on the development of vaccines to induce protective immune responses. Given their admirable safety records, protein subunit vaccines are attractive for widespread immunization, but their disadvantages include poor immunogenicity and expensive manufacture. We show here that engineered Escherichia coli outer membrane vesicles (OMVs) are an easily purified vaccine-delivery system capable of greatly enhancing the immunogenicity of a low-immunogenicity protein antigen without added adjuvants. Using green-fluorescent protein (GFP) as the model subunit antigen, genetic fusion of GFP with the bacterial hemolysin ClyA resulted in a chimeric protein that elicited strong anti-GFP antibody titers in immunized mice, whereas immunization with GFP alone did not elicit such titers. Harnessing the specific secretion of ClyA to OMVs, the ClyA-GFP fusion was found localized in OMVs, resulting in engineered recombinant OMVs. The anti-GFP humoral response in mice immunized with the engineered OMV formulations was indistinguishable from the response to the purified ClyA-GFP fusion protein alone and equal to purified proteins absorbed to aluminum hydroxide, a standard adjuvant. In a major improvement over current practice, engineered OMVs containing ClyA-GFP were easily isolated by ultracentrifugation, effectively eliminating the need for laborious antigen purification from cell-culture expression systems. With the diverse collection of heterologous proteins that can be functionally localized with OMVs when fused with ClyA, this work signals the possibility of OMVs as a robust and tunable technology platform for a new generation of prophylactic and therapeutic vaccines.protein subunit | adjuvant

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Section: Discussionmentioning

confidence: 95%

Section: Discussionmentioning

confidence: 96%

See 1 more Smart Citation

Delivery of foreign antigens by engineered outer membrane vesicle vaccines

Chen

Osterrieder

Metzger

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

256

247

View full text Add to dashboard Cite

show abstract

“…These recombinant Salmonella vaccines can target mucosal inductive tissues (PP) and ultimately disseminate and stimulate both mucosal and systemic arms of the immune system (29,30). It has been shown that the attenuated strain S. Typhi can serve as a safe and effective oral vaccine to prevent typhoid fever and serve as a live vector to deliver heterologous Ags in humans (31)(32)(33)(34). To enhance the immunogenicity of passenger Ag expression by live Salmonella vaccines (35), various types of promoters have been tested to enhance passenger Ag expression.…”

Section: Discussionmentioning

confidence: 99%

Oral Vaccination withSalmonellaSimultaneously ExpressingYersinia pestisF1 and V Antigens Protects against Bubonic and Pneumonic Plague

Yang

Hinnebusch

Trunkle

et al. 2007

The Journal of Immunology

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The gut provides a large area for immunization enabling the development of mucosal and systemic Ab responses. To test whether the protective Ags to Yersinia pestis can be orally delivered, the Y. pestis caf1 operon, encoding the F1-Ag and virulence Ag (V-Ag) were cloned into attenuated Salmonella vaccine vectors. F1-Ag expression was controlled under a promoter from the caf1 operon; two different promoters (P), PtetA in pV3, PphoP in pV4, as well as a chimera of the two in pV55 were tested. F1-Ag was amply expressed; the chimera in the pV55 showed the best V-Ag expression. Oral immunization with Salmonella-F1 elicited elevated secretory (S)-IgA and serum IgG titers, and Salmonella-V-Ag(pV55) elicited much greater S-IgA and serum IgG Ab titers than Salmonella-V-Ag(pV3) or Salmonella-V-Ag(pV4). Hence, a new Salmonella vaccine, Salmonella-(F1+V)Ags, made with a single plasmid containing the caf1 operon and the chimeric promoter for V-Ag allowed the simultaneous expression of F1 capsule and V-Ag. Salmonella-(F1+V)Ags elicited elevated Ab titers similar to their monotypic derivatives. For bubonic plague, mice dosed with Salmonella-(F1+V)Ags and Salmonella-F1-Ag showed similar efficacy (>83% survival) against ∼1000 LD50 Y. pestis. For pneumonic plague, immunized mice required immunity to both F1- and V-Ags because the mice vaccinated with Salmonella-(F1+V)Ags protected against 100 LD50 Y. pestis. These results show that a single Salmonella vaccine can deliver both F1- and V-Ags to effect both systemic and mucosal immune protection against Y. pestis.

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“…There were trials to deliver the antigen using surface display and secretion of ClyA (9,10). In this work, 193-aa S1E epitope from SARSCoV S protein was chosen as a cargo antigen because this has been well characterized as a receptor binding domain (22) and vaccine candidate (14).…”

Section: Discussionmentioning

confidence: 99%

The Bacterial Surface Expression of SARS Viral Epitope using Salmonella typhi Cytolysin A

et al. 2009

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The cytolysin A (ClyA) is a 34 kDa pore-forming cytotoxic protein and expressed by some enteric bacteria including Salmonella typhi. This toxin is transported on the bacterial surface and secreted without posttranslational modification. Using the surface display of ClyA, the expression vectors for 193-aa immunogenic antigen of spike protein (termed S1E) from severe acute respiratory syndrome coronavirus (SARS-CoV) were constructed. The vectors carried a gene encoding S. typhi ClyA conjugated to S1E at the C terminus (termed ClyA-S1E) and asd gene in pGEM-T and pBR322, named pGApLCS1E and pBApLCS1E, respectively. An asd-mutated E. coli transformed with these vectors could grow without diaminopimelic acid (DAP), indicating that they were stably maintained in such mutants. ClyA-S1E recombinant proteins from these vectors were expressed on the surface of the attenuated S. typhimurium deficient of global virulence gene regulator, ppGpp. However, they did not show the hemolytic activity on the blood agar plate and cytotoxicity against HeLa cells. To examine whether bacteria expressing ClyA-S1E induced the immune response against S1E, S. typhimurium deficient of ppGpp and Asd was transformed with these vectors and orally immunized in mice. In the western blotting against GST-conjugated S1E using the immunized mouse sera, it was shown that the significant band was detected in the mouse serum by the bacteria transformed with pGApLCS1E but not with pBApLCS1E. It indicates that the immune response producing antibody was dependent on the expression level of ClyA-S1E. Therefore, ClyA delivery system can be used for SARS vaccine development.

show abstract

Adaptation of the Endogenous Salmonella enterica Serovar Typhi clyA -Encoded Hemolysin for Antigen Export Enhances the Immunogenicity of Anthrax Protective Antigen Domain 4 Expressed by the Attenuated Live-Vector Vaccine Strain CVD 908- htrA

Cited by 65 publications

References 56 publications

Delivery of foreign antigens by engineered outer membrane vesicle vaccines

Delivery of foreign antigens by engineered outer membrane vesicle vaccines

Oral Vaccination withSalmonellaSimultaneously ExpressingYersinia pestisF1 and V Antigens Protects against Bubonic and Pneumonic Plague

The Bacterial Surface Expression of SARS Viral Epitope using Salmonella typhi Cytolysin A

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