A rapid and potent DNA vaccination strategy defined by in vivo monitoring of antigen expression

Bins, Adriaan D.; Jorritsma, Annelies; Wolkers, Monika C.; Hung, Chien Fu; Wu, T. C.; Schumacher, Ton N.; Haanen, John B.A.G.

doi:10.1038/nm1264

Cited by 154 publications

(151 citation statements)

References 20 publications

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“…For this reason, to examine whether hollow microneedle-mediated immunization with PLGA nanoparticles encapsulated OVA may prime OVA-specific endogenous CD8 + T cells in mice, a priming procedure was used consisting of three immunizations delivered over a time period of 6 days. This protocol had been shown to induce cellular immune responses following dermal DNA tattoo immunization [26,42], but not yet when using hollow microneedles [7]. We report here that this prime boost protocol indeed elicits vigorous CD8 + T cell responses in mice immunized with PLGA encapsulated antigen, when using hollow microneedles as delivery method.…”

Section: Discussionmentioning

confidence: 70%

Hollow microneedle-mediated intradermal delivery of model vaccine antigen-loaded PLGA nanoparticles elicits protective T cell-mediated immunity to an intracellular bacterium

Groot

Mönkäre

et al. 2017

Journal of Controlled Release

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A B S T R A C TThe skin is an attractive organ for immunization due to the presence of a large number of epidermal and dermal antigen-presenting cells. Hollow microneedles allow for precise and non-invasive intradermal delivery of vaccines. In this study, ovalbumin (OVA)-loaded poly(lactic-co-glycolic acid) (PLGA) nanoparticles with and without TLR3 agonist poly(I:C) were prepared and administered intradermally by hollow microneedles. The capacity of the PLGA nanoparticles to induce a cytotoxic T cell response, contributing to protection against intracellular pathogens, was examined. We show that a single injection of OVA-loaded PLGA nanoparticles, compared to soluble OVA, primed both adoptively transferred antigen-specific naïve transgenic CD8 + and CD4 + T cells with markedly high efficiency. Applying a triple immunization protocol, PLGA nanoparticles primed also endogenous OVA-specific CD8 + T cells. Immune response, following immunization with in particular anionic PLGA nanoparticles co-encapsulated with OVA and poly(I:C), provided protection against a recombinant strain of the intracellular bacterium Listeria monocytogenes, secreting OVA. Taken together, we show that PLGA nanoparticle formulation is an excellent delivery system for protein antigen into the skin and that protective cellular immune responses can be induced using hollow microneedles for intradermal immunizations.

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Section: Discussionmentioning

confidence: 70%

Hollow microneedle-mediated intradermal delivery of model vaccine antigen-loaded PLGA nanoparticles elicits protective T cell-mediated immunity to an intracellular bacterium

Groot

Mönkäre

et al. 2017

Journal of Controlled Release

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“…Another method that targets cutaneous DCs is the tattooing of DNA. 18,29,31 Using a relatively low amount of DNA (30 mg), tattooing induces strong cellular and humoral responses against antigens driven by ubiquitous promoters like pCMV.…”

Section: Resultsmentioning

confidence: 99%

“…This technique of dermal DNA vaccination has been previously described. 29 DNA vaccinations were performed with 15 mg of pCMV/EGFP as irrelevant DNA, 15 mg of pCMV/OVA or 30 mg of pDC-STAMP/OVA. At day 8, 15 000 OVA-transfected B16OVA tumor cells were injected subcutaneously into the left flank.…”

Section: Transcriptional Targeting Of Dendritic Cells V Moulin Et Almentioning

confidence: 99%

Targeting dendritic cells with antigen via dendritic cell-associated promoters

et al. 2012

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The induction of tumor-specific immune responses is largely dependent on the ability of dendritic cells (DCs) to present tumor-associated antigens to T lymphocytes. Therefore, we investigated the use of DC-associated promoter-driven genetic vaccines to specifically target DC in vivo. Restricted expression of vaccine-encoding genes in DC should enhance specificity and improves their safety for clinical applications. Hereto, 3 --5 kb upstream sequences of the murine genes encoding CD11c, DC-SIGN, DC-STAMP and Langerin were isolated, characterized and subcloned into enhanced green fluorescent protein (EGFP) reporter constructs. Upon electroporation, EGFP was expressed in DC cell lines, but not in other cell lines, confirming DCrestricted promoter activity. When these promoters were cloned into a construct upstream of the gene for ovalbumin (OVA), it appeared that DC-STAMP promoter-driven expression of OVA (pDCSTAMP/OVA) in DC yielded the most efficient OVA-specific CD4 þ and CD8 þ T-cell responses in vitro. Administration of pDC-STAMP/OVA in vivo, using the tattoo gun vaccination system, evoked specific immune responses as evidenced in a mouse tumor model. Adoptively transferred pDC-STAMP/OVA-transfected DCs induced strong CD8 þ T-cell proliferation in vivo. These experiments demonstrate that our DC-directed promoter constructs are potential tools to restrict antigen expression in DC and could be implemented to modulate DC function by the introduction of relevant proteins.

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“…The excellent immunogenicity of dermal DNA vaccination is probably related to the high prevalence of antigen-presenting cells (APCs) in the skin, in the form of Langerhans cells (LCs) in the epidermis and dendritic cells in the dermis (Kanitakis, 2002;Mathers and Larregina, 2006). Several techniques have been developed for the intradermal administration and cellular uptake of naked DNA, such as gene gun and particle injection systems (Klinman et al, 1998;Mitragotri, 2005;Steitz et al, 2006); jet injectors (Mitragotri, 2005;Bahloul et al, 2006); electroporators (Maruyama et al, 2001;Heller et al, 2007;Hooper et al, 2007;Hirao et al, 2008); and a technique, developed at our institute, that has been called ''DNA tattooing'' (Bins et al, 2005;Pokorna et al, 2008). DNA tattooing delivers naked plasmid DNA into the skin through thousands of punctures made with a multiple-needle tattoo device.…”

mentioning

confidence: 99%

“…We have demonstrated that DNA tattooing results in local transfection and expression of the encoded antigen by cells in murine skin. More importantly, the efficacy of DNA tattooing in inducing strong vaccine-specific immune responses has been established in murine models (Bins et al, 2005) and the superiority of DNA tattooing over intramuscular DNA vaccination has been demonstrated in nonhuman primates (Verstrepen et al, 2008).…”

mentioning

confidence: 99%

Optimization of Intradermal Vaccination by DNA Tattooing in Human Skin

et al. 2009

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The intradermal administration of DNA vaccines by tattooing is a promising delivery technique for genetic immunization, with proven high immunogenicity in mice and in nonhuman primates. However, the parameters that result in optimal expression of DNA vaccines that are applied by this strategy to human skin are currently unknown. To address this issue we set up an ex vivo human skin model in which DNA vaccine-induced expression of reporter proteins could be monitored longitudinally. Using this model we demonstrate the following: First, the vast majority of cells that express DNA vaccine-encoded antigen in human skin are formed by epidermal keratinocytes, with only a small fraction (about 1%) of antigen-positive epidermal Langerhans cells. Second, using full randomization of DNA tattoo variables we show that an increase in DNA concentration, needle depth, and tattoo time all significantly increase antigen expression ( p < 0.001), with DNA concentration forming the most critical variable influencing the level of antigen expression. Finally, in spite of the marked immunogenicity of this vaccination method in animal models, transfection efficiency of the technique is shown to be extremely low, estimated at approximately 2 to 2000 out of 1Â10 10 copies of plasmid applied. This finding, coupled with the observed dependency of antigen expression on DNA concentration, suggests that the development of strategies that can enhance in vivo transfection efficacy would be highly valuable. Collectively, this study shows that an ex vivo human skin model can be used to determine the factors that control vaccine-induced antigen expression and define the optimal parameters for the evaluation of DNA tattoo or other dermal delivery techniques in phase 1 clinical trials.

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A rapid and potent DNA vaccination strategy defined by in vivo monitoring of antigen expression

Cited by 154 publications

References 20 publications

Hollow microneedle-mediated intradermal delivery of model vaccine antigen-loaded PLGA nanoparticles elicits protective T cell-mediated immunity to an intracellular bacterium

Hollow microneedle-mediated intradermal delivery of model vaccine antigen-loaded PLGA nanoparticles elicits protective T cell-mediated immunity to an intracellular bacterium

Targeting dendritic cells with antigen via dendritic cell-associated promoters

Optimization of Intradermal Vaccination by DNA Tattooing in Human Skin

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