Insufficient APC Capacities of Dendritic Cells in Gene Gun-Mediated DNA Vaccination

Lauterbach, Henning; Gruber, Anton; Ried, Christine; Cheminay, Cédric; Brocker, Thomas

doi:10.4049/jimmunol.176.8.4600

Cited by 45 publications

(41 citation statements)

References 44 publications

(58 reference statements)

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“…2,3 Promoter activity is also essential for the development of strong immune responses to transfected antigens. Of the other DCassociated promoters that have been described in the literature (fascin, 20 dc-stamp, 23 cd11c, 33 dectin-2 19 and the promoter for murine MHC class II 34 ), the fascin promoter appears the most promising because of its high activity. Fascin promoter constructs are capable of inducing humoral and cellular responses after in vivo gene-gun vaccination to the same extent as those caused by CMV-driven constructs.…”

Section: Discussionmentioning

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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Section: Discussionmentioning

confidence: 99%

Targeting dendritic cells with antigen via dendritic cell-associated promoters

et al. 2012

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“…However, a current challenge of DNA vaccination for eliciting CD8 T cell responses is to increase potency (47). Reduced potency is in large part due to the inefficiency of getting DNA encoded epitopes into the MHC-I antigen presentation pathway and can be dependent upon cross-presentation (52). Reflecting its pre-processed and pre-peptide loaded status, SCT-based DNA vaccination has shown great promise using animal model systems.…”

Section: Discussionmentioning

confidence: 99%

Human Major Histocompatibility Complex (MHC) Class I Molecules with Disulfide Traps Secure Disease-related Antigenic Peptides and Exclude Competitor Peptides

Truscott

Wang

Lybarger

et al. 2008

Journal of Biological Chemistry

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The ongoing discovery of disease-associated epitopes detected by CD8 T cells greatly facilitates peptide-based vaccine approaches and the construction of multimeric soluble recombinant proteins (e.g. tetramers) for isolation and enumeration of antigen-specific CD8 T cells. Related to these outcomes of epitope discovery is the recent demonstration that MHC class I/peptide complexes can be expressed as single chain trimers (SCTs) with peptide, ␤ 2 m and heavy chain connected by linkers to form a single polypeptide chain. Studies using clinically relevant mouse models of human disease have shown that SCTs expressed by DNA vaccination are potent stimulators of cytotoxic T lymphocytes. Their vaccine efficacy has been attributed to the fact that SCTs contain a preprocessed and preloaded peptide that is stably displayed on the cell surface. Although SCTs of HLA class I/peptide complexes have been previously reported, they have not been characterized for biochemical stability or susceptibility to exogenous peptide binding. Here we demonstrate that human SCTs remain almost exclusively intact when expressed in cells and can incorporate a disulfide trap that dramatically excludes the binding of exogenous peptides. The mechanistic and practical applications of these findings for vaccine development and T cell isolation/enumeration are discussed.

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“…þ and CD8 þ cell responses in mice (Lauterbach et al, 2006), suggesting that antigen expression in non-APC cell types may be required for the generation of strong cellular immunity. Thus, antigen expression within keratinocytes is likely to be in large part responsible for the immunogenicity of intradermal DNA vaccines and the measurement of antigen expression in human skin samples is therefore likely to have predictive value.…”

Section: Van Den Berg Et Almentioning

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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Insufficient APC Capacities of Dendritic Cells in Gene Gun-Mediated DNA Vaccination

Cited by 45 publications

References 44 publications

Targeting dendritic cells with antigen via dendritic cell-associated promoters

Targeting dendritic cells with antigen via dendritic cell-associated promoters

Human Major Histocompatibility Complex (MHC) Class I Molecules with Disulfide Traps Secure Disease-related Antigenic Peptides and Exclude Competitor Peptides

Optimization of Intradermal Vaccination by DNA Tattooing in Human Skin

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