We have knowhow on ELISPOT analysis of long peptide vaccines.

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

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

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

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.

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.

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

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

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

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

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.

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.