Generation of large numbers of dendritic cells in a closed system using Cell Factories™

Tuyaerts, Sandra; Noppe, Sofie; Corthals, Jurgen; Breckpot, Karine; Heirman, Carlo; Greef, Catherine De; Riet, Ivan Van; Thielemans, Kris

doi:10.1016/s0022-1759(02)00099-6

Cited by 103 publications

(71 citation statements)

References 42 publications

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“…Both the generation and the cryopreservation of immature DCs have been described previously (41). Briefly, CD14 ϩ monocytes were enriched in a semi-closed culture system using Cell Factories (Nunc).…”

Section: In Vitro Generation Of Dcsmentioning

confidence: 99%

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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Section: In Vitro Generation Of Dcsmentioning

confidence: 99%

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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“…Patients were eligible if their tumor expressed the MAGE-A3 gene and if disease had a locoregional extension (T1-4N2b; TNM (tumor/lymph nodes/metastasis) staging system of the American Joint Committee on Cancer, Chicago, IL) and no visceral extension other than lung metastasis (M1b). The vaccine consisted of 10 million monocyte-derived autologous DC cultured for 6 days in the presence of GM-CSF and IL-4 and matured for 1 day in medium containing IL-4, GM-CSF, TNF-␣, IL-6, IL-1␤, and PGE 2 (14). The HLA-A1-restricted MAGE-A3 peptide EVDPIGHLY 168 -176 (MAGE-3.A1) (15) was added for the last 8 h. Mature DC were collected and pulsed again for 3 h with the MAGE-3.A1 peptide and tetanus toxoid as the recall Ag.…”

Section: Patientmentioning

confidence: 99%

Vaccination of a Melanoma Patient with Mature Dendritic Cells Pulsed with MAGE-3 Peptides Triggers the Activity of Nonvaccine Anti-Tumor Cells

Carrasco

Pel

Neyns

et al. 2008

The Journal of Immunology

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We previously characterized the CTL response of a melanoma patient who experienced tumor regression following vaccination with an ALVAC virus coding for a MAGE-A3 Ag. Whereas anti-vaccine CTL were rare in the blood and inside metastases of this patient, anti-tumor CTL recognizing other tumor Ags, mainly MAGE-C2, were 100 times more frequent in the blood and considerably enriched in metastases following vaccination. In this study we report the analysis of the CTL response of a second melanoma patient who showed a mixed tumor response after vaccination with dendritic cells pulsed with two MAGE-A3 antigenic peptides presented, respectively, by HLA-A1 and HLA-DP4. Anti-MAGE-3.A1 CD8 and anti-MAGE-3.DP4 CD4 T cells became detectable in the blood after vaccination at a frequency of ∼10−5 among the CD8 or CD4 T cells, respectively, and they were slightly enriched in slowly progressing metastases. Additional anti-tumor CTL were present in the blood at a frequency of 2 × 10−4 among the CD8 T cells and, among these, an anti-MAGE-C2 CTL clone was detected only following vaccination and was enriched by >1,000-fold in metastases relative to the blood. The striking similarity of these results with our previous observations further supports the hypothesis that the induction of a few anti-vaccine T cells may prime or restimulate additional anti-tumor T cell clones that are mainly responsible for the tumor regression.

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“…Dendritic cells were isolated as described [28] with minor modifications to increase dendritic cell yields [29]. Briefly, peripheral blood mononuclear cells were isolated from buffy coat preparations of healthy donors by gradient centrifugation (Lymphoprep, Nycomed Pharma AS, Oslo, Norway).…”

Section: Analysis Of Tnfα and Of Nitrite Formationmentioning

confidence: 99%

Human pancreatic duct cells can produce tumour necrosis factor-α that damages neighbouring beta cells and activates dendritic cells

et al. 2004

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Aims/hypothesis. In the human pancreas, a close topographic relationship exists between duct cells and beta cells. This explains the high proportion of duct cells in isolated human islet preparations. We investigated whether human duct cells are a source of TNFα-mediated interactions with beta cells and immune cells. This cytokine has been implicated in the development of autoimmune diabetes in mice. Methods. Human duct cells were isolated from donor pancreases and examined for their ability to produce TNFα following a stress-signalling pathway. Ductcell-released TNFα was tested for its in vitro effects on survival of human beta cells and on activation of human dendritic cells. Results. Exposure of human pancreatic duct cells to interleukin-1β (IL-1β) induces TNFα gene expression, synthesis of the 26,000 M r TNFα precursor and conversion to the 17,000 M r mature form, which is rapidly released. This effect is NO-independent and involves p38 MAPK and NF-κB signalling. Duct-cellreleased TNFα contributed to cytokine-induced apoptosis of isolated human beta cells. It also induced activation of human dendritic cells. Conclusions/interpretation. Human pancreatic duct cells are a potential source of TNFα that can cause apoptosis of neighbouring beta cells and initiate an immune response through activation of dendritic cells. They may thus actively participate in inflammatory and immune processes that threaten beta cells during development of diabetes or after human islet cell grafts have been implanted.

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Generation of large numbers of dendritic cells in a closed system using Cell Factories™

Cited by 103 publications

References 42 publications

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

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

Vaccination of a Melanoma Patient with Mature Dendritic Cells Pulsed with MAGE-3 Peptides Triggers the Activity of Nonvaccine Anti-Tumor Cells

Human pancreatic duct cells can produce tumour necrosis factor-α that damages neighbouring beta cells and activates dendritic cells

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