A functional analysis of anti-MAGE-3.A1 CTL clones derived from vaccinated patients (either ALVAC canarypox virus vector, peptide-pulsed DC or peptide vaccination +/- montanide adjuvant) who displayed tumor regression was done. The functional avidities of these CTL clones, evaluated in lysis assays, were surprisingly low, suggesting that high avidity was not part of the putative capability of these CTL to trigger tumor rejection. Most anti-MAGE-3.A1 CTL clones obtained after DC vaccination, but not after peptide or ALVAC vaccination, produced IL-10. Transcri...

Post-vaccination frequencies of the T cells of anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 were found to be >= 10-fold higher than that of pre-vaccinations, indicating a specific CTL response to these vaccinations. An analysis performed on PBL collected after the 3 vaccinations of cycle 2 clearly shows that the frequencies for both anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 T cells are still elevated.

Direct administration of ovalbumin (OVA) encoding lentiviral vectors caused in vivo transduction of cells that were found in draining lymph nodes (LNs) and induced potent anti-OVA cytotoxic T cells similar to those elicited by ex vivo transduced DC.

Post-vaccination frequencies of the T cells of anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 were found to be >= 10-fold higher than that of pre-vaccinations, indicating a specific CTL response to these vaccinations. An analysis performed on PBL collected after the 3 vaccinations of cycle 2 clearly shows that the frequencies for both anti-NY-ESO-1.A2 and anti-Tyrosinase.A2 T cells are still elevated.

After using frozen MoDC for tumor peptide specific stimulation of autologous T lymphocytes at a ratio of 1 DC per 10 T cells, co-cultures led to antigen specific, IFN-? secreting T cells (1 restimulation at a DC:T cell ratio of 1:20).

We investigated the interactions between human monocyte derived DCs (MDDC), generated in the presence of GM-CSF and IL-4 (IL-4 DC), and antigen-stimulated circulating gamma delta T lymphocytes, bearing the Vgamma2 TCR. The studies demonstrated for the first time the existence of a bidirectional activating interaction between DCs and gamma delta T lymphocytes activated by aminobiphosphonates. These data suggest a potential adjuvant role of this early cross-talk in the therapeutic activity of aminobiphosphonate drug that are currently used as anti-tumor drugs.

These T cells were generated from the Leukapheresis product, after elutration, of a metastatic lesion of stage III/IV melanoma patients, expanded by co-culture with tumor lysate loaded DC.

The autologous T lymphocytes were stimulated with (frozen) MoDC at a ratio of 1 DC per 10 T cells.

B cell proliferation (polyclonal, oligoclonal, monoclonal) was observed in human Hemato-Lymphoid System Rag2-/-gc-/- mice after being infected with EBV and mounting an immune response.

To quantify presentation of the epitopes by DC, we used bulk T cells electroporated with TCR-encoding RNA in stimulation assays.

Testing the improved RCC product PME-CD40L DC it is noteworthy that presence of CD28+ T cells in tumor lymphocytic infiltrates correlates with favorable clinical outcome.

We studied the correlation between CD8 multiple cell surface markers and functional profiles studied at single cell level. For that purpose, we subdivided peripheral CD8 T cells into eleven different cell subtypes based on the association of multiple cell surface markers. In each subtype, we isolated single-cells. In each single cell, we quantified the expression of multiple genes. Moreover, we isolated and studied cells from different normal donors.

These T cells were associated with multiple markers into 14 different cell types to identify subsets. The findings thus describe new CD8+ cell subsets, allow the identification of relatively homogeneous CD8+ subpopulations, provide a predictable and precise correlation between particular cell surface markers and CD8+ T-cell functional properties, and identify effector cells present in both CCR7-CD45RA+ and CCR7-CD45R0+ compartments. The results also indicate that activated cells might modulate the expression of CD45RA/R0 asynchronously rather than CCR7-CD45...

We observed that ICAM-1 expression by mature DCs is critical for long-lasting contacts with CD8+ T cells, but dispensable for short-lived antigen-specific interactions. Serial brief T cell-dendritic cell contacts induced early CD8+ T cell activation, proliferation and effector CTL differentiation in the first few days after immunization.

CEA specific CD4+ T cells were found both in normal donors and in patients with high-grade cervical lesions, pancreas adenocarcinoma and advanced melanoma but CD4+ T cells from the patients compared to normal donors showed impaired IFN-gamma production.

Our current efforts in a study of functional homogeinity involve a transition to 8 colours surface staining, and the evaluation of antigen-specific cell sets identified with MHC tetramers. For the later process, we initiated data collection of EBV, CMV and HIV specific T cell sets.

Cytotoxic T cells proliferation was observed in human Hemato-Lymphoid System Rag2-/-gc-/- mice after being infected with EBV and mounting an immune response.

Our current efforts in a study of functional homogeinity involve a transition to 8 colours surface staining, and the evaluation of antigen-specific cell sets identified with MHC tetramers. For the later process, we initiated data collection of EBV, CMV and HIV specific T cell sets.

These T cells are specific for the P14-GP33 antigen. They were used to study the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

These T cells are specific for the 0T-1-OVA antigen. They were used to study the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.