title: Transcriptional Targeting of Mature Dendritic Cells with Adenoviral Vectors via a Modular Promoter System for Antigen Expression and Functional Manipulation.
abstract: To specifically target dendritic cells (DCs) to simultaneously express different therapeutic transgenes for inducing immune responses against tumors, we used a combined promoter system of adenoviral vectors. We selected a 216 bp short Hsp70B' core promoter induced by a mutated, constitutively active heat shock factor (mHSF) 1 to drive strong gene expression of therapeutic transgenes MelanA, BclxL, and IL-12p70 in HeLa cells, as well as in mature DCs (mDCs). As this involves overexpressing mHSF1, we first evaluated the resulting effects on DCs regarding upregulation of heat shock proteins and maturation markers, toxicity, cytokine profile, and capacity to induce antigen-specific CD8(+) T cells. Second, we generated the two-vector-based "modular promoter" system, where one vector contains the mHSF1 under the control of the human CD83 promoter, which is specifically active only in DCs and after maturation. mHSF1, in turn, activates the Hsp70B' core promotor-driven expression of transgenes MelanA and IL-12p70 in the DC-like cell line XS52 and in human mature and hence immunogenic DCs, but not in tolerogenic immature DCs. These in vitro experiments provide the basis for an in vivo targeting of mature DCs for the expression of multiple transgenes. Therefore, this modular promoter system represents a promising tool for future DC-based immunotherapies in vivo.
authors.affiliation: Department of Immune Modulation at the Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstrasse 14, 91052 Erlangen, Germany.
authors.forename: Ilka
authors.identifier: 0000-0002-2866-3901
authors.initials: I
authors.lastname: Knippertz
authors.affiliation: Department of Immune Modulation at the Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstrasse 14, 91052 Erlangen, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Deinzer
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstrasse 14, 91052 Erlangen, Germany.
authors.forename: Jan
authors.identifier: 0000-0002-3478-0741
authors.initials: J
authors.lastname: Dörrie
authors.affiliation: Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstrasse 14, 91052 Erlangen, Germany.
authors.forename: Niels
authors.identifier: 0000-0001-8236-9298
authors.initials: N
authors.lastname: Schaft
authors.affiliation: German Cancer Research Center, Im Neuenheimer Feld 280, 69120 Heidelberg, Germany.
authors.forename: Dirk M
authors.identifier: 0000-0002-1225-2593
authors.initials: DM
authors.lastname: Nettelbeck
authors.affiliation: Department of Immune Modulation at the Department of Dermatology, Universitätsklinikum Erlangen, Hartmannstrasse 14, 91052 Erlangen, Germany.
authors.forename: Alexander
authors.identifier: 0000-0001-7187-6729
authors.initials: A
authors.lastname: Steinkasserer
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D015703:Antigens, CD / Q000235:genetics; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D002454:Cell Differentiation; D016207:Cytokines / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D003713:Dendritic Cells / Q000276:immunology / Q000502:physiology*; D005822:Genetic Vectors; D006367:HeLa Cells; D000076249:Heat Shock Transcription Factors; D006801:Humans; D007136:Immunoglobulins / Q000235:genetics; D018664:Interleukin-12 / Q000235:genetics; D058965:MART-1 Antigen / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D011401:Promoter Regions, Genetic; D014157:Transcription Factors / Q000235:genetics*; D019076:Transgenes; D051020:bcl-X Protein / Q000235:genetics; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D016207:Cytokines; D004268:DNA-Binding Proteins; D000076249:Heat Shock Transcription Factors; D007136:Immunoglobulins; D018664:Interleukin-12; D058965:MART-1 Antigen; D008562:Membrane Glycoproteins; D014157:Transcription Factors; D051020:bcl-X Protein; D000097802:CD83 Antigen; C494811:BCL2L1 protein, human