DNA immunization: Effects of vehicle and route of administration on the induction of protective antiviral immunity

Yokoyama, Masayuki; Zhang, Jie; Whitton, J. Lindsay

doi:10.1111/j.1574-695x.1996.tb00290.x

Cited by 37 publications

(10 citation statements)

References 35 publications

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“…For example, while injection of pCMV-NP leads to long-term antigen expression and to immunity, it is possible that the immunity is induced almost immediately following injection, despite the immaturity of the neonatal immune system, and that long-term antigen expression is unnecessary. It is also unclear why only 48% of the mice responded to the immunization, but these data are very similar to those obtained in previously published studies on adult DNA immunization against LCMV, in which around 50 to 75% of mice responded (20,(35)(36)(37)(38). In light of the strong similarity in the rates of successful vaccination in adults and neonates, it appears unlikely that the vaccination failures in the present study reflect a specific defect in the ability to immunize young animals.…”

Section: Discussionsupporting

confidence: 83%

“…Infection induces a variety of antiviral effector mechanisms, but protective immunity requires the presence of virus-specific major histocompatibility complex class I-restricted CD8 ϩ T lymphocytes (9,15,16,31,39,40). Immunization with LCMV nucleoprotein (NP), expressed by recombinant vaccinia virus or plasmid DNA, can induce anti-LCMV CTL which confer protection on adult mice (31,36,37). Although antibody does not seem to play a vital role in virus clearance, maternally derived anti-LCMV antibodies can protect pups (3); however, this passive protection is transient and is greatly diminished by 5 weeks of age (2 weeks postweaning) (3).…”

mentioning

confidence: 99%

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Neonatal DNA immunization with a plasmid encoding an internal viral protein is effective in the presence of maternal antibodies and protects against subsequent viral challenge

Hassett

Zhang

Whitton

1997

J Virol

108

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Conventional vaccines are remarkably effective in adults but are much less successful in the very young, who are less able to initiate a mature immune response and who may carry maternal antibodies which inactivate standard vaccines. We set out to determine whether DNA immunization might circumvent these problems. We have previously shown that intramuscular injection of plasmid DNA encoding the nucleoprotein (NP) gene of lymphocytic choriomeningitis virus (LCMV) is capable of inducing immune responses and protecting 50% of adult mice against lethal and sublethal challenge with LCMV. Here we demonstrate that mouse pups injected with the same plasmid hours or days after birth produce major histocompatibility complex-restricted, NPspecific cytotoxic T lymphocytes (CTL) that persist into adulthood; 48% of vaccinated pups responded to subsequent sublethal viral challenge by the accelerated production of anti-NP LCMV-specific CTL, indicating that these animals had been successfully immunized by the plasmid DNA. In addition, these mice showed a >95% reduction in splenic viral titers 4 days postinfection compared to control mice, demonstrating a more rapid control of infection in vivo. Furthermore, pups born of and suckled on LCMV-immune dams (and therefore containing passively acquired anti-LCMV antibodies at the time of DNA inoculation) responded to the DNA vaccine in a similar manner, showing that maternally derived anti-LCMV antibodies do not significantly inhibit the generation of protective immune responses following DNA vaccination. These findings suggest that, at least in this model system, DNA immunization circumvents many of the problems associated with neonatal immunization.

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Section: Discussionsupporting

confidence: 83%

mentioning

confidence: 99%

Neonatal DNA immunization with a plasmid encoding an internal viral protein is effective in the presence of maternal antibodies and protects against subsequent viral challenge

Hassett

Zhang

Whitton

1997

J Virol

108

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“…As shown in Fig. 4, pCMV-NP induced anti-LCMV antibodies, as we have previously demonstrated (39,40); all 16 mice responded. Remarkably, in both mouse strains, pCMV-U-NP failed to induce anti-LCMV an-tibodies; the ELISA titers of mice immunized with this plasmid were indistinguishable from those of mice immunized with a plasmid encoding ubiquitin alone.…”

Section: Fig 2 Cotranslational Ubiquitination Enhances Degradation supporting

confidence: 78%

“…6. Compared to mice immunized either with saline or with pCMV-U alone, mice immunized with pCMV-NP showed significant (2 to 3 logs) reduction in virus titers, as we have previously noted (37,39,40). Nevertheless, virus was still detectable in all eight pCMV-NP-immunized mice.…”

Section: Fig 2 Cotranslational Ubiquitination Enhances Degradation supporting

confidence: 61%

DNA immunization: ubiquitination of a viral protein enhances cytotoxic T-lymphocyte induction and antiviral protection but abrogates antibody induction

Rodrı́guez

Zhang

Whitton

1997

J Virol

248

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DNA immunization can induce cytotoxic T lymphocytes (CTL), antibodies, and protection against microbial challenge. The underlying mechanisms remain obscure and must be understood to permit rational manipulation and optimization of the technique. We set out to enhance the intracellular degradation of a viral antigen, with the intent of improving antigen entry into, and presentation by, the class I major histocompatibility complex pathway. We achieved this goal by cotranslational ubiquitination of a plasmid-encoded viral antigen, lymphocytic choriomeningitis virus (LCMV) nucleoprotein (NP). We show that native NP is very stable in cell culture, while the ubiquitinated product is so rapidly degraded that it is barely detectable. This rapid degradation leads to more efficient sensitization of target cells in an in vitro cytotoxicity assay, consistent with enhanced antigen presentation, and both degradation and target cell recognition are blocked by a proteasome inhibitor. We have used the plasmid for in vivo studies and find that, remarkably, ubiquitination leads to a complete abrogation of antibody responses, presumably because the encoded protein is so rapidly and completely degraded that insufficient antigen remains to interact appropriately with B cells. In contrast, in vivo CTL induction is improved by ubiquitination of NP. That CTL are induced at all by this rapidly degraded protein may shed light on the mechanism by which CTL are induced by DNA immunization; it has been suggested that CTL induction following intramuscular DNA injection results not from antigen presentation by cells taking up and expressing the DNA but rather from uptake of soluble protein by specialized antigenpresenting cells (APC). It appears to us unlikely that the ubiquitinated protein could function in this manner, since it is so rapidly degraded in vitro and fails to induce antibodies in vivo. Finally, the ubiquitinated protein confers markedly enhanced protection against LCMV challenge. Mice immunized with a plasmid encoding NP show approximately 100-fold reductions in virus titers compared to controls, while mice immunized with a plasmid encoding the ubiquitinated NP show reductions in virus load of at least 5 ؋ 10 4-to 5 ؋ 10 5-fold. This is by far the most effective DNA vaccine that we have yet designed. Ubiquitination therefore may improve DNA immunization, but caution is warranted, since immunity to many microbes depends on induction of good humoral immunity, and we show here that this may be prevented by ubiquitination of the encoded protein.

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“…Protection is CTL° mediated and does not depend on the induction of antiviral antibodies. The vehicle (saline, or lipid-associated) and the route of administration are important in determining the level of induced immunity (Yokoyama et al, 1996;Yokoyama et al, 1997). The LCMV model has allowed the demonstration of the exquisite flexibility of DNA vaccines.…”

Section: A Dna Vaccines Against Lcmvmentioning

confidence: 99%

DNA immunization and central nervous system viral infection

Whitton

Fujinami

2001

Advances in Virus Research

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DNA immunization: Effects of vehicle and route of administration on the induction of protective antiviral immunity

Cited by 37 publications

References 35 publications

Neonatal DNA immunization with a plasmid encoding an internal viral protein is effective in the presence of maternal antibodies and protects against subsequent viral challenge

Neonatal DNA immunization with a plasmid encoding an internal viral protein is effective in the presence of maternal antibodies and protects against subsequent viral challenge

DNA immunization: ubiquitination of a viral protein enhances cytotoxic T-lymphocyte induction and antiviral protection but abrogates antibody induction

DNA immunization and central nervous system viral infection

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