In our laboratory we carry out multiplex assays. For instance, multiplex analysis was used to study the response of human MoDC to yeast, spheroplasts, pseudohyphae and spores.

We have experience in western blotting, which we use for example for the assessment of the efficiency of gene silencing.

We have expertise in western blotting. Western blot was used, for instance, for the demonstration of protein kinase R phosphorylation on transduction.

In our lab we have experience in performing real time PCR analyses.

This puromycin-based technology was developed in our laboratory to monitor translation by FACs in individual or cell populations. It is a non-invasive method to monitor protein synthesis and cellular activation in single cells or heterologous populations.

Our lab has expertise in performing thymidine-incorporation assays. The assay is used, for instance, for the monitoring of the vaccine induced specific anti-tumor T cell responses.

We have know-how on TaqMan gene expression assays by Perkin-Elmer Applied Biosystems. They are used, for instance, for PCR analysis.

For quantitative proteomic studies, our group recently described the SILAC (Stable Isotope Labelling by Amino Acids in Cell Culture) approach. In this method, independent populations of cells are grown in medium containing different non-radioactive isotope forms of essential amino acids (e.g. arginine and lysine). Because the cells cannot synthesize these amino acids, after some generations all proteins are labelled and therefore can be distinguished and quantified by the mass spectrometer. To generate a proteomic map of DCs which includes quantitative information that...

The method is about transfecting CD4(+) and CD8(+) T cells with mRNA encoding recombinant immunoreceptors for use in the adoptive immunotherapy of cancer. CD4(+) and CD8(+) T cells were efficiently transfected with immunoreceptors specific for ErbB2 and CEA. The immunoreceptor expression was transient with half-maximal expression at day 2 and no detectable immunoreceptor expression at day 9 after electroporation. Opposed to standard procedures using retroviral gene transduction to constitutively express immunoreceptors in T cells, transient immunoreceptor expressi...

We have expertise on an HSV-based technology for the delivery of antigens to dendritic cells.

In our work, we have experience in applying peptide-tetramer assays. Primary and secondary CTL response against the C8 T cell epitope SIINFEKL was monitored in a study for maturing myeloid DCs and plasmacytoid DCs.

Real time and immunohistochemistry analysis was performed in a study to evaluate whether iNOS can serve as a prognostic factor in stage III melanoma survival. The analysis showed that both iNOS and COX-2 alone significantly predicted OS. The BRAF/NRAS mutation status did not significantly differ between patient groups, although iNOS significantly correlated with BRAF mutation frequency. Furthermore, iNOS and COX-2 were significantly linked to a number of metastatic lymph nodes. However, iNOS maintained its statistical significance together with a number of lymph node...

Using MAGE-A3 as a model antigen we established a functional read-out system by transfection of CD4 T cells with RNA encoding T cell receptors recognizing MHC DP4 restricted MAGE-A3 epitopes.

Using this peptide library technique and ELISPOT analysis patients´ PBMC are pre-screened for CD4- and CD8- T cell responses and individual sets of 4 to 8 peptides chosen for further analysis.

In our lab, we have know-how about performing MPLC assays, such as using it in the establishment of immunomonitoring vaccination trials with RNA transfected DC by using overlapping peptides.

Our lab has expertise in lipofection of tumor-antigen-encoding RNA into DC. The method was compared to RNA electroporation into DC.

The laboratory has expertise in the use of MACS anti-CD14 microbeads by Miltenyi Biotec, Bergisch-Gladbach, Germany. They were used for isolation of monocytes from PBMCs.

We have know-how about performing LDA assays. We have, for instance, monitored vaccine induced T cell responses for the two clinical trials. We found CD8+ and CD4+ T cell responses against the immunological tracers KLH and TK, and against the NTPs and MAGE-3, respectively.

We have know-how about LDA assays; for instance, we use LDA assays to monitor vaccine induced T cell responses.

We have developed immunological techniques for monitoring of tumour antigen specific CD4+ T cell responses. We set an in vitro system able to detect fona fide in vivo primed CD4+ T cells. Purified CD4+ T cells are first stimulated in vitro once in the presence peptides, previously identified as containing naturally processed epitopes on tumour-associated antigens. After two weeks, proliferation and cytokine secretion (IFN-?, IL-5, IL-4, IL-2, TNF-?, IL-10, TGF-?, IL-17, granzyme and perforin) are tested. To estimate the frequency of antigen specific CD4+ T cell we us...