Induction of antigen-specific CD8+ cytotoxic T cells by dendritic cells co-electroporated with a dsRNA analogue and tumor antigen mRNA

Michiels, Annelies; Breckpot, Karine; Corthals, Jurgen; Tuyaerts, Sandra; Bonehill, Aude; Heirman, Carlo; Thielemans, Kris; Aerts, JL

doi:10.1038/sj.gt.3302750

Cited by 29 publications

(25 citation statements)

References 40 publications

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“…[15][16][17] In spite of the shared endpoints, the outcome of dsRNA treatment is dependent on cell type 15,30 and the method of exposure. 18,19,21,31 Indeed, in our study, we demonstrate that it is necessary to transfect dsRNA into the cytoplasm of AML cells in order to exert its immunostimulatory effects. These data corroborate the previously published data on dsRNA treatment of thyrocytes or murine non-plasmacytoid DC.…”

Section: Discussionmentioning

confidence: 99%

“…Diebold et al 18 showed that murine non-plasmacytoid DC secrete high levels of IFN-a in response to dsRNA, only when the latter was introduced into the cytoplasm to mimic direct viral infection. For human DC, Michiels et al 19 reported that higher numbers of Melan-A-specific cytotoxic T lymphocytes (CTL) were obtained following stimulation with Melan-A mRNA-loaded DC that were co-electroporated with poly(I:C 12 U), a clinical-grade dsRNA analog, as compared to DC matured by an inflammatory cytokine cocktail. In vivo mouse studies have shown that vaccination with dsRNAelectroporated tumor cells enhances the antitumor activity via TLR3 activation in the endosomes of mouse DC, resulting in a protective antitumor response mediated by cell-associated but not by soluble dsRNA.…”

Section: Introductionmentioning

confidence: 99%

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Proinflammatory response of human leukemic cells to dsRNA transfection linked to activation of dendritic cells

et al. 2007

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Leukemic cells exert immunosuppressive effects that interfere with dendritic cell (DC) function and hamper effective antileukemic immune responses. Here, we sought to enhance the immunogenicity of leukemic cells by loading them with the double-stranded (ds) RNA Toll-like receptor 3 (TLR3) ligand polyriboinosinic polyribocytidylic acid (poly(I:C)), mimicking viral infection of the tumor cells. Given the responsiveness of DC to TLR ligands, we hypothesized that the uptake of poly(I:C)-loaded leukemic cells by immature DC (iDC) would lead to DC activation. Primary acute myeloid leukemia (AML) cells and AML cell lines markedly responded to poly(I:C) electroporation by apoptosis, upregulation of TLR3 expression, enhanced expression of major histocompatibility complex (MHC) and costimulatory molecules and by production of type I interferons (IFN). Upon phagocytosis of poly(I:C)-electroporated AML cells, DC maturation and activation were induced as judged by an increased expression of MHC and costimulatory molecules, production of proinflammatory cytokines and an increase of T helper 1 (T H 1)-polarizing capacity. These immune effects were suboptimal when AML cells were passively pulsed with poly(I:C), indicating the superiority of poly(I:C) transfection over pulsing. Our results demonstrate that poly(I:C) electroporation is a promising strategy to increase the immunogenicity of AML cells and to convert iDC into activated mature DC following the phagocytosis of AML cells.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Proinflammatory response of human leukemic cells to dsRNA transfection linked to activation of dendritic cells

et al. 2007

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“…We previously demonstrated that electroporation is a powerful technique for the delivery of both TAA (mRNA) and activation stimulus (poly(I:C 12 U)) to DCs (11). It has also been demonstrated that similar electroporation conditions can be applied for the introduction of siRNA into DCs (46).…”

Section: Electroporation Of Dcs With Melan-a/mart-1 Encoding Mrna Pomentioning

confidence: 99%

“…Electroporation of siRNA into DCs. Immature DCs were coelectroporated with Melan-A/MART-1 mRNA, poly(I:C 12 U) and siRNA by an optimized protocol (11). Briefly, after two wash steps, 4 ϫ 10 6 DCs were resuspended in a final volume of 200 l Optimix Solution B (EQUIBIO, Ashford, U.K.) containing 20 g mRNA, 1 g poly(I:C 12 U) and 120 pmol siRNA.…”

Section: Transfection Of DC With Sirnamentioning

confidence: 99%

“…It has been shown that TLR ligands are necessary and can synergize to fully activate DCs to overcome tolerogenic mechanisms as well as active inhibition by regulatory CD4 ϩ T cells (Treg), factors which impede a productive anti-tumor immune response (5)(6)(7)(8)(9)(10). In this regard, it has been demonstrated that DCs matured with the TLR3 ligand poly(I:C) or its clinical grade analog poly(I:C 12 U) are potent inducers of TAA-specific CTLs (11,12).…”

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Attenuated Expression of A20 Markedly Increases the Efficacy of Double-Stranded RNA-Activated Dendritic Cells As an Anti-Cancer Vaccine

Breckpot

Aerts-Toegaert

Heirman

et al. 2009

The Journal of Immunology

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A20 is a zinc finger protein with ubiquitin-modifying activity. A20 has been described as negatively regulating signaling induced by the TNF receptor and TLR family in a number of cell types, including mouse bone marrow-derived dendritic cells (DCs). However, the expression and effect of A20 in activated human monocyte-derived DCs have not been previously evaluated. We report that DCs activated with the TLR3 ligand poly(I:C) up-regulate A20. Down-regulating A20 demonstrated its role in the functional activation of DCs. A20 down-regulated DCs showed higher activation of the transcription factors NF-κB and activator protein-1, which resulted in increased and sustained production of IL-6, IL-10, and IL-12p70. We additionally silenced the immunosuppressive cytokine IL-10 and demonstrated that IL-10 inhibits T cell proliferation. We further demonstrated that A20 down-regulated DCs skew naive CD4+ T cells toward IFN-γ producing Th1 cells, a process which is dependent on IL-12p70 and which is unaffected by IL-10. Furthermore, A20 and/or IL-10 down-regulated DCs had an enhanced capacity to prime Melan-A/MART-1 specific CD8+ T cells. Finally, we demonstrated that potent T cell stimulatory DCs are generated by the simultaneous delivery of poly(I:C12U), A20, or A20/IL-10 small interfering RNA and Ag-encoding mRNA, introducing a one step approach to improve DC-based vaccines. Together these findings demonstrate that A20 negatively regulates NF-κB and activator protein-1 in DCs and that down-regulation of A20 results in DCs with enhanced T cell stimulatory capacity.

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Targeting Toll-Like Receptor for the Induction of Immune and Antitumor Responses

Lustgarten

Hoelzinger²,

Duque³

et al. 2009

Targeted Cancer Immune Therapy

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Induction of antigen-specific CD8+ cytotoxic T cells by dendritic cells co-electroporated with a dsRNA analogue and tumor antigen mRNA

Cited by 29 publications

References 40 publications

Proinflammatory response of human leukemic cells to dsRNA transfection linked to activation of dendritic cells

Proinflammatory response of human leukemic cells to dsRNA transfection linked to activation of dendritic cells

Attenuated Expression of A20 Markedly Increases the Efficacy of Double-Stranded RNA-Activated Dendritic Cells As an Anti-Cancer Vaccine

Targeting Toll-Like Receptor for the Induction of Immune and Antitumor Responses

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