The flow cytometer at the disposal of the laboratory is used, for example, to assess Th subsets.

The flow cytofluorometer available to the laboratory is used, for example, for the assessment of differentiation, maturation, and intracellular cytokine production of DCs.

The FACS analyser available to the laboratory is used, for instance, to analyse stained cells.

In our labs we have experience in performing immunohistochemistry, e.g., for studying the cellular organization of the marrow of long bones.

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

This flow cytometer was used eg., for an analysis of CD11c populations in the lung and spleen during acute aerosol MTb infection.

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.

We have experience with the exponentially modified Protein Abundance Index (emPAI) method, a label free spectral counting procedure for determining relative protein amounts of LC-MS/MS data. A comprehensive protein inventory of clinical grade immature and cytokine cocktail matured (Il-6, IL-1 beta, TNF alpha, PGE2; 48 hours) monocyte derived human dendritic cells (DC) from a healthy donor has been established by using high accuracy, high sensitivity protein identification technology. The criteria applied resulted in a list of 2794 unique proteins that were identifie...

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.

A comprehensive protein inventory of clinical grade immature and cytokine cocktail matured (Il-6, IL-1 beta, TNF alpha, PGE2; 48 hours) monocyte derived human dendritic cells (DC) from a healthy donor has been established. To this aim, 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.

The integral plasma membrane proteome of D1 cells stimulated for 24 hours with LPS has been analyzed with this LC-MS/MS analyser and compared to the results obtained from immature D1 cells.

The Challenge of Cancer: Individual Cancers and Mutations Every individual´s cancer is unique. Therefore, the most effective treatment is one customized to a particular cancer´s genetic configuration. Genetic mutations and chromosomal abnormalities are commonly associated with human cancers. Unfortunately, since the genetic mutations leading to the development of cancer are often random events, every patient's tumor can contain a unique repertoire of antigens. This characteristic of human cancer requires that each patient's immune system be able to recognize and ...

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.

An assay for measuring cytotoxic T lymphocytes was used for the monitoring of T cell activation in the evaluation of the immunotherapy potential of DC pulsed with tumor antigens.

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.

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

There is a GMP facility for conduction of phase III clinical trials available. The structural up-grade of this facility has limited the number of patients recruited.

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

The gene gun system available to the laboratory was used, e.g., for micro-particle bombardment of murine skin.