A Luminex flow cytometer is available to our lab. It is used, among other things, to quantify the TNFalpha and IFNgamma secretion in a study on antigen recognition.

There is a Luminex flow cytometer available in the lab. It is used, among other things, for assaying of cytokines in B-CLL patients treated with Apo-DC +/- GM-CSF.

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.

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

We have developed a MAGE-ML export feature in collaboration with the European Bioinformatics Institute (EBI).

Iterative calibration algorithms were used to achieve a final, absolute mass accuracy of better than 10 parts per million (p.p.m.) for all peptide ions. These high-accuracy spectra were searched against a human protein database, using MASCOT probability-based scoring in which the fragment ions are matched against the calculated fragments of all tryptic peptides from the human sequences. Additional constraints were set by searches against a reversed human database to minimize false positive protein identifications.

Our laboratory has a mass spectrometer at its disposal.

Mass spectrometry (MS)-based proteomics has become a powerful technology to map the protein composition of organelles, cell types and tissues. In our department, a large-scale effort to map these proteomes is complemented by the Max-Planck Unified (MAPU) proteome database. MAPU contains several body fluid proteomes; including plasma, urine, and cerebrospinal fluid. Cell lines have been mapped to a depth of several thousand proteins and the red blood cell proteome has also been analyzed in depth. The liver proteome is represented with 3200 proteins. By employing hi...

The MaxQuant software is designed to make it possible to compare cell subsets quantitatively based on the extracted ion current (XIC) and without any need of stable isotope labels.

The microarray device at our disposal uses Affymetrix GeneChip® technology.

The lab has a midi-MACS magnetic cell sorting device by Miltenyi Biotec, Bergisch-Gladbach, Germany, that is used, e.g., to isolate monocytes from PBMC.

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 an MRI scanner at its disposal. We use it, e.g., to track dendritic cells in melanoma patients for monitoring of cellular therapy.

The MRI facility available in the laboratory is used, among other things, to image vaccines.

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.

We have experience in multicolour flow cytometry. Besides ELISPOT, tetramer based 8-colour flow cytometry has become our routine monitoring assay, allowing extended phenotypic and functional characterization of T cell subpopulations. Of special interest has become the identification of polyfunctional T cells (capable to simultaneously exert several functions, i.e. production of various cytokines and killing activity) since these have been shown to elicit more effective immune responses in HIV vaccination trials.

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.

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.

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.

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...