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.

This gene gun was used for the delivery of vectors engineered to express distinct chemokines at the local skin site of mice.

A Zeiss LSM 510 confocal microscope is available for use in the lab.

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.

The yeast oligonucleotide array was constructed using the S. cerevisiae Genome Oligo Set ™ (Operon Technologies, CA, USA) composed of 6240 optimized oligonucleotides (70mers) each representing one yeast gene.

In our lab we have a two-photon microscope. For instance, cell death was assessed in the viable epidermis by non-invasive near infrared two-photon microscopy following micro-particle bombardment of murine skin.

There is a two-photon intravital microscope available in the laboratory. It was used, among other things, to analyze the infiltration and destruction of solid tumors by CTLs.

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...

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

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

This spectrometer was, among other things, used for an analysis of the integral plasma membrane proteome of D1 cells.

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

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.

Microarray and gene expression are fundamental tools of nowadays Biology research. While standard formats and software systems have been developed to represent and publicly share information about microarray experiments, existing computational solutions give limited support to the representation of the outcomes, experimental hypotheses or conclusions about biological questions that are dealt with in gene expression analysis. We propose an OWL-based model and a Semantic Web-based approach to address the issue. We show that the formalization of microarray-related knowle...

Our lab has know-how in the functional annotation of upregulated and downregulated genes by gene ontology classification and pathway analysis by several bioinformatics approaches, among them Ontologizer. Preliminary results obtained from querying DAVID (http://david.abcc.ncifcrf.gov/) and Ontologizer (http://www.charite.de/ch/medgen/ontologizer/) show statistically significant enrichment for immunological-relevant signaling pathways and immunology-related GO terms of the upregulated proteins in mature DCs.

This platform with its state-of-the-art mass spectrometry facilities is led by Mann (P33). To enhance the platform, two additional groups were recruited as Third Parties: P114 Edwin Lasonder in Nijmegen (NL), and P113 Juri Rappsilber in Edinburgh (GB). The mass spectrometer has been used, for instance, to identify and quantify chemokine dependent tyrosine phosphorylation events. Furthermore, the success of SILAC labelling was analysed using the mass spectrometer.

This tool is available through the DC-THERA Directory (see "Pathway Signature Repository "). The procedure allows a user to compare his/her microarray data to the corpus of public immunology experiments stored in DC-BASE using pathway signatures. The procedure is made up by several steps. 1.Raw data normalization: the user uploads raw data (Affymetrix or Illumina gene expression platforms are supported) and select the species. Upon succesful upload, raw data is normalized using a standard procedure. 2.Comparison selection: The user is then asked to select which sa...