We have experience with ELISA for IL-12p70 from R&D System.

An ELISA assay using the kit from Biosource was performed according to the manufacturer’s instructions for the measurement of different cytokines.

Our lab has expertise with Elispot assays. We evaluated the capacity of autologous DC and HEK293 cells transfected with relevant HLA alleles to function as T cell targets in Elispot assays upon transfection with tumor antigen encoding plasmids.

We have know-how on assays to measure the proportions of Treg cells. For instance, we set up an assay to measure the proportions of Treg cells in tissues or cells that is based on the demethylation of a region of the FOXP3 promoter, which is observed in human Treg cells but not in the other T cells that can express FOXP3 transiently after activation. Starting from DNA, two quantitative PCR are used, which amplify the methylated and unmethylated FOXP3 promoter sequences. The ratio between the expression levels is, in our hands, a faithful Treg marker. We will pursue...

In march 2009 we obtained the new BD ARIA II, which not only allows 14 parameter flow cytometry, but also sorting of even very small antigen specific T cell subpopulations for further molecular analysis in close collaboration with Pierre Coulie´s group in Brussels. [PhD student Steve Voland]

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.

We have a confocal microscope available. This microscope was is used, for instance, for the characterization of antigen depots.

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

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...

An Illumina transcriptional profiling platform is available for microarray analyses.

We have been developing the DC-ATLAS DC-pathway database. For this, we have set about identifying key signalling pathways that were likely to be of most relevance to dendritic cell functions. We have designed a DC-ATLAS data model to effectively describe pathways, their genes, and their interaction. We produced a controlled vocabulary that contains at least 400 ontological terms, prepared on the basis of the proposals and discussions among the researchers involved in DC-ATLAS project. We defined the pathway representation where each pathway was described using three ...

The experiments on human monocyte-derived DC are being annotated in DC-BASE, a unique DC-dedicated database integrated with data mining tools. The database uses a LIMS system based on the structure of Base2.9. The database currently contains 240 microarray experiments annotated accordingly to MIAME standards and thoroughly analyzed in order to pass severe quality control tests. A meeting will be organized for defining policies on data sharing within the DC-THERA community and beyond. In order to populate DC-BASE with data obtained from the literature, we used the ...

The GenePix 4000 software package was used to quantify microarray fluorescence in a yeast microarray experiment.

A GenePix 4000 microarray scanner is available in our lab (manufacturer: Axon, Foster City, CA).

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.

We have expertise in performing ELISA assays. For instance, we use the ELISA kit for IL-12 and IL-10 from Biosource to evaluate cytokine accumulation in supernatants at 24h, according to a standard protocol and it was measured at 450n

The FAX analyser available in the laboratory is used, e.g., to measure transfection efficiency.

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

There is a MACS separator (Miltenyi Biotech) at the disposal of our lab. It is used, for example, to purify CD8 T cells.

In the lab, there is a multi-photon microscope available. It is used, for instance, in the analysis of in vivo distribution and motility of NK cells.