This DC vaccine was produced from matured DC (matured using TNF alpha, IFN alpha and PGE2) which were subsequently transfected with CD40L-encoding RNA and tumor RNA.

After 6 hour of stimulation, DCs were collected, washed 3 times with PBS, treated with zymolyase, washed twice and cells, lysated with a hypotonic solution (KCl 0.05%), were plated on YPD. Survival of yeast cells, spores or hyphae after uptake was reported as percentage of colony forming units after 3 days relative to the total number of cells growing in the absence of DCs exposure. To assess the importance of ROS production in DC killing ability, DPI (10 microM) was added 30 minutes before stimulation and survival of microorganisms was assessed using the same...

Attenuated Expression of A20 Markedly Increases the Efficacy of Double Stranded RNA-Activated Dendritic Cells as an Anti-Cancer Vaccine A20 is a zinc finger protein with ubiquitine-modifying activity. It has been described that A20 negatively regulates signaling induced by the tumor necrosis factor receptor and toll like receptor (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 monocyte-derived DCs have not been previously evaluated. We report that DC...

The dead cells were stained and used in the uptake of dead cells by DC.

DCs pre-loaded in vitro with ICs are at least 1000-fold more potent than ICs injected directly into mice or DCs loaded with the same amount of non-complexed protein. FcgammaRs on DCs were required for efficient priming of Ag-specific CD8+ cells in vivo and induction of tumor protection.

We showed that, in the absence of adjuvants, DEX mediate potent antigen dependent-antitumor effects against preestablished tumors in mice pretreated with immunopotentiating dosing of cyclophosphamide (CTX). CTX could i) abolish the suppressive function of CD4+CD25+Foxp3+ regulatory T cells (Treg), ii) markedly enhance the magnitude of secondary but not primary CTL responsesinduced by DEX vaccines, iii) synergize with DEX in therapy but not prophylaxis tumor models.

These marginal zone dendritic cells, interacting with the cholera toxin (CT) adjuvant, lead to effective priming of an immune response in vivo. For the first time, we have followed adjuvant targeting of MZ DC in vivo.

This DC vaccine was produced from the Leukapheresis product, after elutriation, of a metastatic lesion of stage III/IV melanoma patients.

The receptors of these DC were blocked by addition of laminarin, mannan and chitin.

These dendritic cells were obtained from PBMC from which monocytes had been isolated that differentiated into DC after cell culture.

Constructs for fusion proteins of the DC specific receptor DC-SIGN fused to GFP have been completed and tested.

These DC were electroporated, co-cultured with purified CD8 T cells using MACS cell separation to asses the in vitro CD8 T cell stimulatory capacity of DC.

Autologous dendritic cells are loaded with apoptotic leukemic cells from patients with B-CLL to be used as vaccine in patients with chronic lymphocytic leukemia. Preliminary data from a clinical study indicate reduction of circulating tumor cells. No adverse effects have been observed.

Three patients were enrolled in a clinical trial and DC vaccine was produced. The DC vaccine was administered into irradiated tumours in RCC of one patient only due to cancer progression in the other two patients. The patient treated is in complete remission 6 months after treatment.

This vaccine was used for RCC patients who were deficient in T cell IFN-gamma and IL-2 production pre-vaccination.

We also show that DCs derived from ashen mice, which are defective for the small GTPase Rab27a, fail to cross present antigens efficiently, due to increased phagosome acidification and antigen degradation. This defect in Rab27a-deficient DCs results from the impaired recruitment to phagosomes of the NOX2 membrane components. Therefore phagosomal alkalinization by NOX2 is controlled by Rab27a, and is required for efficient cross presentation in DCs.

100 micrograms/ml curdlan (Wako) was used (along with R848, LPS, yeast RNA and yeast cells in different conditions of culture) to induce DC activation.

The cDNA’s encoding these early expressed HIV antigens have been human codon optimized and modified with lysosomal targeting sequences.

Monocytes were cultured in RPMI 1640 medium (GIBCO BRL) supplemented with 2 mM L-glutamine (Sigma), 1% (vol/vol) non-essential amino acids, 100 mM sodium pyruvate, 50 U/ml of penicillin and 50 mg/ml of streptomycin (Gibco BRL) containing 10% (vol/vol) FCS (Hyclone).

We have prepared several new constructs for the expression of (modified) adhesion receptors in DC.