Chitin (500 micrograms/ml) was added to DCs in order to block their beta-glucan receptors.

Mannan (500 micrograms/ml) was added to DCs in order to block their mannan receptors.

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...

A long-standing collaboration with Centre for Ecology & Hydrology (CEH, UK) has led to the identification of an activity in salivary gland extracts of the tick Rhipicephalus appendiculatus that profoundly modulates the responses of human monocyte-DC towards extrinsic stimuli. The gene for this molecule, which we have termed Japanin, has been cloned and recombinant protein has been generated. This material inhibits the up-regulation of costimulatory molecules in response to certain TLR agonists, and some cytokines but not others, and modulates the function of...

A long-standing collaboration with Centre for Ecology & Hydrology (CEH, UK) has led to the identification of an activity in salivary gland extracts of the tick Rhipicephalus appendiculatus that profoundly modulates the responses of human monocyte-DC towards extrinsic stimuli. Recombinant protein has been generated from the gene for this molecule (Japanin) which had been cloned. This material inhibits the up-regulation of costimulatory molecules in response to certain TLR agonists, and some cytokines but not others, and modulates the function of the DC in ...

Manufacturing of cGMP RNA to be used as an API (active pharmaceutical ingredient) in the planned Tri-Mix DC RNA Trial will be performed after obtaining the manufacturing license for RNA loaded DCs.

To assess the importance of ROS production in DC killing ability, 2-(3,4-Dihydroxyphenylimino)-imidazolidine (10 microM) was added to the dendritic cells 30 minutes before stimulation.

Zymolyase was used to treat DC, which were used in a study of microrganism survival following uptake by DCs.

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

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

10 mg/ml sulfo-NHS-LC-biotin (Sigma-Aldrich) in 50 mM NaHCO3 pH 8.5 were used to biotinylate yeasts / spores for 2 hours at 4°C. The remaining reactive biotin molecules were inactivated by incubation in 100 mM Tris-HCl pH 8.0 for 40 minutes at 4°C. DCs were then treated with biotinylated spores/yeasts.

This vaccine was used in patients with colorectal cancer or ovarium cancer. It was well tolerated and robust p53-specific T cell responses were induced.

DMSO was used to dissolve blue formazan particles.

2M KOH were used to dissolve blue formazan particles.

DCs were exposed to cytochalasin D (10 microg/ml, TebuBio) for 30 minutes at 4°C.

In a spore culture, zymolyase (2 mg/ml) was used to digest ascum and liberate spores. We initially performed experiments to evaluate zymoliase sensitivity of asci for this specific strain and we assessed that 15 minutes incubation was the minimum time needed to damage the ascus, without touching the spore cell wall. Zymoliase was inactivated by heat (65°C for 2 minutes) and washed away carefully together with the remainings of the empty ascus, by resuspending twice the spore culture in 500 µl of distilled water centrifuging and discarding the supernatant; th...

There is siRNA available in the lab.

There are lentiviral vectors available in the lab.

We have shown recently that the T cell stimulatory capacity of DCs pulsed with tumorantigen-derived peptides can be considerably increased by activating the DCs through electroporation with CD40L, CD70 and constitutively active TLR4 encoding mRNA (TriMix DCs). This vaccine will also be tested in the multicentre trial.

We have chemically coupled the S-epimer of the Pam3CSK4 to long peptides containing a CTL epitope and used this structure to investigate the behaviour of the diastereomer.