Salmonella typhimurium delta aroA delta aroD mutants expressing a foreign recombinant protein induce specific major histocompatibility complex class I-restricted cytotoxic T lymphocytes in mice

Turner, Stephen J.; Carbone, Federico; Strugnell, R A

doi:10.1128/iai.61.12.5374-5380.1993

Cited by 55 publications

(36 citation statements)

References 36 publications

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“…This strain induced the highest levels of B cell immunogenicity (Table 4 and Figures 2B, 3 and 4). This result is similar to those reported in previous studies, which have found that lowering the plasmid copy number results in enhanced plasmid stability and a reduction in the expression levels of heterologous antigens to non-toxic levels [31][32][33][34].…”

Section: B Cell Responses and Soluble Antigen Expression Levelssupporting

confidence: 92%

Comparative immunological evaluation of recombinant Salmonella Typhimurium strains expressing model antigens as live oral vaccines

Zheng

Zhang

et al. 2012

BMC Immunol

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BackgroundDespite the development of various systems to generate live recombinant Salmonella Typhimurium vaccine strains, little work has been performed to systematically evaluate and compare their relative immunogenicity. Such information would provide invaluable guidance for the future rational design of live recombinant Salmonella oral vaccines.ResultTo compare vaccine strains encoded with different antigen delivery and expression strategies, a series of recombinant Salmonella Typhimurium strains were constructed that expressed either the enhanced green fluorescent protein (EGFP) or a fragment of the hemagglutinin (HA) protein from the H5N1 influenza virus, as model antigens. The antigens were expressed from the chromosome, from high or low-copy plasmids, or encoded on a eukaryotic expression plasmid. Antigens were targeted for expression in either the cytoplasm or the outer membrane. Combinations of strategies were employed to evaluate the efficacy of combined delivery/expression approaches. After investigating in vitro and in vivo antigen expression, growth and infection abilities; the immunogenicity of the constructed recombinant Salmonella strains was evaluated in mice. Using the soluble model antigen EGFP, our results indicated that vaccine strains with high and stable antigen expression exhibited high B cell responses, whilst eukaryotic expression or colonization with good construct stability was critical for T cell responses. For the insoluble model antigen HA, an outer membrane expression strategy induced better B cell and T cell responses than a cytoplasmic strategy. Most notably, the combination of two different expression strategies did not increase the immune response elicited.ConclusionThrough systematically evaluating and comparing the immunogenicity of the constructed recombinant Salmonella strains in mice, we identified their respective advantages and deleterious or synergistic effects. Different construction strategies were optimally-required for soluble versus insoluble forms of the protein antigens. If an antigen, such as EGFP, is soluble and expressed at high levels, a low-copy plasmid-cytoplasmic expression strategy is recommended; since it provokes the highest B cell responses and also induces good T cell responses. If a T cell response is preferred, a eukaryotic expression plasmid or a chromosome-based, cytoplasmic-expression strategy is more effective. For insoluble antigens such as HA, an outer membrane expression strategy is recommended.

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Section: B Cell Responses and Soluble Antigen Expression Levelssupporting

confidence: 92%

Comparative immunological evaluation of recombinant Salmonella Typhimurium strains expressing model antigens as live oral vaccines

Zheng

Zhang

et al. 2012

BMC Immunol

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“…The pIC20H plasmid, which is derived from pUC19, has the highest copy number of the plasmids tested and it is possible that the increased copy number may have contributed to the plasmid's instability. A relationship between plasmid instability and the copy number has been observed in previous studies [24,31]. Turner et al [31] reported that ovalbumin expressed from a high-copy-number plasmid delivered by attenuated S. typhimurium was unable to induce ova-speci¢c cytotoxic T lymphocytes (CTLs) whereas the same antigen encoded on a lowercopy-number plasmid induced CTLs.…”

Section: Discussionmentioning

confidence: 80%

“…The titre of antibody present in mouse sera were determined using a standard endpoint ELISA. Immunoplates (Nunc A/S, Kamstrup, Denmark) were either coated with 10 Wg ml 31 S. typhimurium lipopolysaccharide (LPS ; Sigma, St Louis, MO, USA) in PBS, or 2 Lf (£occulating units) ml 31 tetanus toxoid (CSL Ltd, Parkville, Vic., Australia) in carbonate coating bu¡er (pH 9.6). Bound antibody was detected by adding sheep anti-mouse immunoglobulin (Ig) HRP conjugate (Silenus Laboratories, Hawthorn, Vic., Australia).…”

Section: Measurement Of Antibody Response By Enzyme-linked Immunosorbmentioning

confidence: 99%

In vitro and in vivo stability of recombinant plasmids in a vaccine strain ofSalmonella entericavar. Typhimurium

Dunstan

Simmons

Strugnell

2003

FEMS Immunology &Amp; Medical Microbiology

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This study examined the ability of different plasmid vectors encoding H(C) fragment, the non-toxic binding portion of tetanus toxin, to be stably retained by Salmonella enterica var. Typhimurium (Salmonella typhimurium) vaccine strain BRD509 and, upon immunisation, to induce an antibody response against the carried antigen. The H(C) fragment expression cassette containing the transcription/translation signals, H(C) fragment open reading frame and the downstream TrpA terminator, was excised from pTETtac4 and incorporated into the plasmids pIC20H, pBR322, pACYC184 and pRSF1010. The resulting constructs were transferred into attenuated S. typhimurium, BRD509, and the level of H(C) fragment expression was examined by Western blot analysis. The relative stability of each plasmid in S. typhimurium was determined in vitro in the absence of antibiotic selection, and in vivo following immunisation. The ability of each H(C) fragment-expressing strain to induce lipopolysaccharide- and tetanus toxoid-specific antibody responses was assayed by an enzyme-linked immunosorbent assay. These studies showed that all the vaccine vector constructs, except the S. typhimurium carrying the expression vector based on pIC20H, were able to elicit a high titre immune response. The level of tetanus toxoid-specific antibody induced by S. typhimurium directly correlated with the level of in vitro and in vivo stability of the H(C) fragment expression plasmid carried by the bacterium, and not with an increased copy number of the parent plasmid vector.

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“…Stable expression of antigen gene is extremely important for oral vaccine using attenuated Salmonella, as it relates to whether it could constantly effectively stimulate the body to produce an immune response. Studies affirmed that stability of plasmid might be relevant to the plasmid copy number [6], and ovalbumin expressed by an attenuated Salmonella harboring a low-copy number plasmid could stimulate to produce specific anti-ovalbumin (CTLs reaction) when carrying a high-copy number plasmid cannot [29]. In order to enhance plasmid stability during immunization, antibiotics were injected into the mice with the period of immunity, yet the result revealed that it is not practical to get the plasmids enhanced in vivo by this method [27].…”

Section: Discussionmentioning

confidence: 99%

Immunogenicity of transmissible gastroenteritis virus (TGEV) M gene delivered by attenuated Salmonella typhimurium in mice

et al. 2016

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Attenuated Salmonella typhimurium (S. typhimurium) was selected as a transgenic vehicle for the development of live mucosal vaccines against transmissible gastroenteritis virus (TGEV) based on the M gene. An approximate 1.0 kb DNA fragment, encoding for glycoprotein M, was amplified by RT-PCR and cloned into eukaryotic expression vector pVAX1. The recombinant plasmid pVAX-M was transformed by electroporation into attenuated S. typhimurium SL7207, and the expression and translation of the pVAX-M delivered by recombinant S. typhimurium SL7207 (pVAX-M) was detected both in vitro and in vivo. BALB/c mice were inoculated orally with SL7207 (pVAX-M) at different dosages to evaluate safety of the vaccines. The bacterium was safe to mice at a dosage of 2 × 10(9) CFU, almost eliminated from the spleen and liver at week 4 post-immunization and eventually cleared at week 6. Mice immunized with 1 × 10(9) CFU of SL7207 (pVAX-M) elicited specific anti-TGEV local mucosal and humoral responses including levels of IgA, IgG, IL-4, and IFN-γ as measured by indirect ELISA assay. Moreover, the control groups (pVAX group, PBS group) maintained at a normal level during week 4-8 post-immunization. The results indicated that attenuated S. typhimurium could be used as a delivery vector for oral immunization of TGEV M gene vaccine.

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Salmonella typhimurium delta aroA delta aroD mutants expressing a foreign recombinant protein induce specific major histocompatibility complex class I-restricted cytotoxic T lymphocytes in mice

Cited by 55 publications

References 36 publications

Comparative immunological evaluation of recombinant Salmonella Typhimurium strains expressing model antigens as live oral vaccines

Comparative immunological evaluation of recombinant Salmonella Typhimurium strains expressing model antigens as live oral vaccines

In vitro and in vivo stability of recombinant plasmids in a vaccine strain ofSalmonella entericavar. Typhimurium

Immunogenicity of transmissible gastroenteritis virus (TGEV) M gene delivered by attenuated Salmonella typhimurium in mice

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