Three groups of end stage cervical cancer patients (in total N=35) were subcutaneously vaccinated with HPV16 E6 combined with or separated from HPV16 E7 overlapping long peptides in Montanide ISA-51 adjuvant, 4 times at three week intervals. Group 1 received 300 microgram per peptide at a single site, group 2 received 100 microgram per peptide of the E6 peptides in one limb, and 300 microgram per peptide of the E7 peptides in a second limb. Group 3 received separate injections of E6 and E7 peptides, each at a dose of 50 micrograms per peptide. The primary endpo...

In the process of extracting total RNA from yeast cells, the pellets were washed with 70% Ethanol two times.

Total RNA was extracted from yeast cells after buffering, incubation and centrifugation by phenol extraction.

These monocytes were extracted from a Leukapheresis product from cancer patients using elutriation.

The FC-block was used for incubation of cells (1:50 in PBS + 2 % FCS) for FACS staining of activation markers.

These mice were generated with the intention to elucidate in vivo DC developmental regions in steady state and inflammation in situ.

These T cells are specific for the 0T-1-OVA antigen. They were used to study the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

These T cells are specific for the P14-GP33 antigen. They were used to study the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

Exosomes were used to treat a chohort of 77 gastrointestinal stromal tumor (GIST) cancer patients.

Expression vectors for GFP-RIG-I used to analyse the interaction between RIG-I, a viral sensing protein, and NS1, an inhibitor of interferon production that is encoded by influenza A virus have been made available.

Fabs that bind strongly to L-SIGN, but to a lesser degree or not at all to dendritic cell-specific ICAM-grabbing nonintegrin (DC-SIGN) were isolated and characterized. We believe that the isolated Abs may be useful for selective delivery of Ags to DC-SIGN- or L-SIGN-bearing APCs for the modulation of immune responses and for blocking viral infections.

We found a novel M-CSF dependent DC developmental pathway that is independent of Flt3L. These DC have unique characteristics and precursor origin. The cells can be found in vivo in Flt3L gene deleted (-/-) mice injected with recombinant M-CSF.

The model antigen OVA was fused to the C1C2 exosome-binding domain of MFG-E8/lactadherin. This construct was used in a study on the capture of antigen-bearing exosomes by DCs and cross-presentation of tumour antigen in exosomes.

Our current efforts in a study of functional homogeinity involve a transition to 8 colours surface staining, and the evaluation of antigen-specific cell sets identified with MHC tetramers. For the later process, we initiated data collection of EBV, CMV and HIV specific T cell sets.

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.

These DCs have been generated from monocytes after elutriation and have not been pulsed with irradiated tumor cells (apoptotic bodies).

The crude population of DC is divided into the three known splenic subsets (CD8+, CD4+, DN DC) using Fluorescence activated cell sorting (BD FACS Aria). This procedure allowed to obtain highly pure fractions of each subset. However, because DC are a very rare population, this method is very material- and cost-intensive. Furthermore, it is not surprising that the obtained cell numbers of each subset after isolation are below 107. However, these cell numbers should be sufficient for proteomic investigations. The proteomes of all three subsets using mass spectrometry is in progress.

The tumor antigen derived peptide pulsed monocyte-derived DC were diluted in medium containing 20 ng/mL TNF-?, plus IL-4 and GM-CSF.

DMSO was used to dissolve blue formazan particles.

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