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

Our laboratory has expertise in applying a fluorescence microscopy method to analyse proximal signalling events induced by cross-linking C-type lectins.

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.

We have experience about using Elispot assays for monitoring vaccine induced specific anti-tumor T cell responses.

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...

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 knowhow on ELISPOT analysis of long peptide vaccines.

An Elispot assay was applied in the establishment of immunomonitoring vaccination trials with RNA transfected DC by using overlapping peptides.

In our laboratory we have know-how on Elispot analysis. For instance, an IFN-ELISPOT assay was used to determine that co-cultures of DC and T cells led to antigen specific, IFN-? secreting T cells after 1 restimulation at a DC:T cell ratio of 1:20.

Knowledge on ELISPOT analyses is available in our lab for immunomonitoring of patients treated with Apo-DC +/- GM-CSF.

Our lab has experience in carrying out ELISPOT analyses. For instance, an IFNgamma ELISPOT assay with PBMC loaded with peptide pools (each consisting of 10 15-mers, which overlap with 11 aminoacids) was done for a study on antigen recognition.

Our lab has expertise about ELISPOT analyses from the long peptide trial in end stage cervical carnoma, resected cervical carcinoma and VIN III patients.

We have experience with ELISA assay for IL-17 from e-Bioscience.

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

Our lab has expertise in performing ELISA assays. For example, we have monitored vaccine induced T cell responses for 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.

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

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

These bioassays were designed taking advantage of some results obtained by performing global gene expression analyses. We have optimised a DC-based assay aimed at identifying new adjuvant molecules potentially capable to induce Th1 responses. This bioassay is a list of standardized procedures, which enables to use the dendritic cells as tools for the determination of the effects of new chemical compounds on dendritic cell capacity to produce type I IFNs.

This bioassay is a list of standardized procedures, which enables to use the dendritic cells as tools for the determination of the effects of new chemical compounds on dendritic cell capacity to produce type I IFNs. The selected molecules could be potential new vaccine adjuvants.