Our lab has know-how in performing 3H-thymidine-incorporation assays. This assay was used in the monitoring of vaccine induced T cell responses.

In our laboratory we perform 51Cr-release assays, for instance in the monitoring of vaccine induced T cell responses.

We have expertise in performing toll-like receptor (TLR)-driven luciferase reporter assays. For example, in a study on the activation of DC we applied a TLR-driven luciferase reporter assay which showed dose-dependent activation of TLR2, TLR3, and TLR8, which was independent of the pseudotype, production, or transduction protocol and was abrogated on heat inactivation.

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 laboratory, we have experience in performing CBA assays. We use them, e.g., in the monitoring of vaccine induced T cell responses.

There is know-how on classical MLR assays in our lab. They are used, for instance, for characterization of silenced cells.

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.

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.

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.

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

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 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.

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

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

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

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

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

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.

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

We have knowhow on ELISPOT analysis of long peptide vaccines.