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

We have expertise in DNA vaccination with the gene-gun available to us to directly transfect skin DC in vivo either with expression plasmids or siRNA plasmids to knockdown target genes and follow the effect on the resulting T cell response.

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

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

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

The Genopolis microarray database serves as an easy interface for the data generated by the Affymetrix GeneChip platform. DC transcriptomic data produced in the past 4 years has made been an accessible resource for data sharing and data integration accessible to all the DC community. The experiments on mice and rat are currently being annotated and conserved in the Genopolis database. Direct access to the primary data held in the Biopolo database is responsibility of the BIOPOLO Project Management System (see http://www.biopolo.it/).

"The ArrayExpress Tab2MAGE package consists of a set of scripts and modules which can read in microarray experiment annotation in defined spreadsheet formats, combine it with a set of experiment data files, and output the result in MAGE-ML format. Tools are included for validation of these spreadsheet documents, and automation of the entire process. This package can handle two distinct spreadsheet formats: MAGE-TAB, the new community-designed spreadsheet format for microarray data representation and interchange, and the eponymous Tab2MAGE, an older, simpler format u...

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.

An Illumina transcriptional profiling platform is available for microarray analyses.

We have experience on performing immunofluorescence assays. As an example, an assay on ordered sequential sections was used to analyze the infiltration and destruction of solid tumors by CTLs.

Experience has been gained in identifying DC presenting a specific peptide via a sensitive immunofluorescent staining technique.

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

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...

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.

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

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 access to and experience with using a linear ion trap / Fourier Transform (LTQ-FT) hybrid mass spectrometer. For example, we use it to analyse purified tryptic peptides.

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

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. We have identified 2794 proteins in DCs by liquid chromatography tandem mass spectrometry.