Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD4 surface markers (and additionally, CD8, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

These surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with surface markers (such as CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.

The Pharmacological compound 1?,25-Dihydroxyvitamin D3 [1,25(OH)2D3], the biologically active metabolite of Vitamin D3, and its analogues, was evaluated for its effect on the DC differentiation/activation pathway induced by type I IFN. The suppressive effect of 1,25(OH)2D3 was associated with a potent impairment of DC migration in response to inflammatory and lymph node homing chemokines, thus unraveling a novel mechanism involved in 1,25(OH)2D3-mediated immunomodulation.

These dendritic cells were generated from adherent peripheral blood monocytes, subject to quality controls, and subsequently used for intradermal administration in HLA-A1 and HLA-A2 positive patients with stage III/IV melanoma (V administrations with eventual additional ones depending on clinical outcome).

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.

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

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.

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.

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.

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.

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.

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

2 ml of acid phenol were added to the buffered and incubated S. cerevisia cells after transferral of the supernatant to a new tube.

One volume fresh medium containing 2x concentrated cytokines was added to the cell culture of monocytes in serum free medium in the presence of 100 ng/mL GM-CSF (Leukine, Berlex) and 20 ng/mL IL-4 (Cell Genix).

This construct was used as in the transfection of immature DC.

The vaccine is composed by autologous DCs electroporated with mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3).