We evaluated the capacity of autologous DC and HEK293 cells transfected with relevant HLA alleles to function as T cell targets in Elispot assays upon transfection with tumor antigen encoding plasmids. Due to strong background from the autologous DC we decided that HEK293 transfected with one HLA-allele at a time plus simultaneously transfected with up to 5 tumor antiges would be optimal to screen for antigen specificity in the patients T cells. However, during a second T cell culture to expand large numbers of T cells for this screening procedure a drastic ex...

We are collecting ascitic fluid (and blood) of ovarian cancer patients to study the presence of immature myeloid populations as compared to healthy donor blood. We have identified two candidate populations that will be tested for the ability to suppress T cells.

Autologous DC were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap to produce the advanced prostate carcinoma vaccine.

These carcinoma cells were used to pulse autologous DCs for vaccine production for ovarian carcinoma patients.

APC-labeled streptavidin was used to detect intracellular yeasts/spores.

The autologous, apoptotic, leukemic cells (Apo-DC) vaccine was given to 2 cohorts of patients: without additional adjuvants and with GM-CSF as an adjuvant. These two cohorts were monitored for 52 weeks. The clinical and immune monitoring data is being currently analyzed.

These patients receive a DC vaccine composed by autologous DCs electroporated with mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3).

BruCells is developing allogeneic dendritic cell fusion vaccines as a therapeutic cancer immune therapy targeted at treating glioma or renal cell carcinoma (RCC). The active substance consists of hybrid cells resulting from the fusion of allogeneic dendritic cells (DC) with allogeneic glioma or RCC tumour cells. Therefore the fusion vaccine qualifies as a somatic cell therapy medicinal product, as defined in Part IV (Advanced therapy medicinal products) of annex I to Directive 2001/83/EC (1), as amended by Directive 2003/63/EC (2). (1) Directive 2001/83/EC...

aHLA-DR fluorescein isothiocyanate was used to label yeast and spores.

It was found that injection of (presumably agonistic) anti-CTLA-4 mAbs induced regulatory T cells.

Natural regulatory T cells (CD4+ CD25+) were depleted by injection of anti-CD25 mAbs, whereas regulatory T cells were induced by injection of (presumably agonistic) anti-CTLA-4 mAbs.

When identifying molecular signatures for alternatively activated DC (AA-DC) it was found that the hallmark of AA-DC was the production of the angiogenic cytokine VEGF in vitro, and in vivo when the cells were implanted into the chicken embryo CAM assay. VEGF production by DC was selectively observed only when the DC where alternatively activated. Therefore, VEGF production by DC can be considered a signature of this state of activation.

Direct administration of ovalbumin (OVA) encoding lentiviral vectors caused in vivo transduction of cells that were found in draining lymph nodes (LNs) and induced potent anti-OVA cytotoxic T cells similar to those elicited by ex vivo transduced DC.

The agonists were used as adjuvants in vivo to induce DC activation and CD4+ T cell and antibody responses.

In our effort to demonstrate that pathogen-derived signals to DC mediated via TLRs can be modulated by activated iNKT cells, DC isolated from animals treated simultaneously with TLR and iNKT cell ligands were potent stimulators of naive T cells in vitro compared with these DC from animals treated with the ligands individually.

In our effort to demonstrate that pathogen-derived signals to DC mediated via TLRs can be modulated by activated iNKT cells, these DC isolated from animals treated simultaneously with TLR and iNKT cell ligands were potent stimulators of naive T cells in vitro compared with DC from animals treated with the ligands individually.

This newly engineered antibody specific for alpha-galactosylceramide (alpha-GalCer)-human CD1d (hCD1d) complexes was used to measure the affinity of binding of iNKT TCR to hCD1d molecules loaded with a panel of alpha-GalCer analogues and to assess the rate of dissociation of alpha-GalCer and alpha-GalCer analogues from hCD1d molecules.

The vaccine is composed by autologous DCs pulsed with apoptotic allogenic prostate carcinoma cell line LNCap. It is used for vaccination of patients with advanced prostate carcinoma.

A functional analysis of anti-MAGE-3.A1 CTL clones derived from vaccinated patients (either ALVAC canarypox virus vector, peptide-pulsed DC or peptide vaccination +/- montanide adjuvant) who displayed tumor regression was done. The functional avidities of these CTL clones, evaluated in lysis assays, were surprisingly low, suggesting that high avidity was not part of the putative capability of these CTL to trigger tumor rejection. Most anti-MAGE-3.A1 CTL clones obtained after DC vaccination, but not after peptide or ALVAC vaccination, produced IL-10. Transcri...

Post-vaccination frequencies of the T cells of anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 were found to be >= 10-fold higher than that of pre-vaccinations, indicating a specific CTL response to these vaccinations. An analysis performed on PBL collected after the 3 vaccinations of cycle 2 clearly shows that the frequencies for both anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 T cells are still elevated.