These are strains of Saccharomyces cerevisiae that were cultured and collected in different conditions: - different growth phase (exponential and stationary phase) - different growth media (standard and promoting pseudohyphal growth) - different cell form (spheroplast, spore and whole cell)

These constructs were prepared for a per-clinical model of breast cancer.

Monocyte-derived DCs were added to the yeast at a final concentration of 5x105 cells/ml into 96-well plates.

In a process of total RNA extraction from S. cerevisia, the cells were collected and the pellet were resuspend in 3.6 ml AE Buffer.

These K/O mice were studied with regard to responses to viral infections. We used the mouse pox virus Ectromelia and MVA-BN.

The immunotherapy potential of DC, pulsed with tumor antigens, was evaluated in EG7-OVA and P815 tumor models. Depletion of natural regulatory T cells in these tumor bearing mice resulted in rejection of P815 cells, but not EG7-OVA, suggesting that regulatory T cells have a stronger impact on immune responses against weakly immunogenic or auto-antigen (P1A).

In order to identify the APCs that were responsible for the development of this state of immune tolerance, we aimed to generate a transgenic mouse that express a recombinant fluorescent protein in the mammary glands, an approach that should result in high levels of expression of the fluorescent protein in the milk. We thus should be able to detect the cells that capture, degrade and present this antigen to T cells. Since we had previously generated a monoclonal antibody, 2C44, that reacts to a peptide derived from the Leishmania LACK protein bound to I-Ad MHC ...

These mice were used to study « in vivo » responses to Ags. We studied the kinetics of effector genes expression and association in TCR-Tg CD8 cells responding to the male antigen and to Listeria-OVA.

We demonstrated using a syngeneic mouse tumor model, expressing an Ag derived from the early region 1A of human adenovirus type 5, that the inadequate nature of the antitumor CTL response is not due to direct Ag presentation by the tumor cells, but results from presentation of tumor-derived Ag by nonactivated CD11c(+) APC.

These patients receive DC transfected with RNA encoding MelanA, Mage3 and Survivin +/- E/L-selectin by the i.v. route. 2nd phase of the DERMA-ER-DC 06 trial.

These stage III/IV melanoma patients underwent surgical removal of a metastatic lesion and lymphodepletion prior to vaccination and adoptive transfer of autologous T cells.

Serotype A C. albicans strain SC5314 was cultured overnight in Saboraud medium at 28°C and then shifted to RPMI medium for 18 hours at 37°C to allow hyphal growth.

S. cerevisiae strain BY4741 och1 (Mata his3delta1 leu2delta0 met15delta0 ura3delta0 OCH1::kanMX4) was cultured in complete medium till exponentially phase. The cellswere biotinylated, labeled and - after treatment of DCs - detected using APC-labeled streptavidin and analyzed by flow cytometry.

S. cerevisiae strains BY4741 (genotype, Mata his3delta1 leu2delta0 met15delta0 ura3delta0) was cultured in complete medium till exponentially phase. The cells were biotinylated, labeled and - after treatment of DCs - detected using APC-labeled streptavidin and analyzed by flow cytometry.

Saccharomyces cerevisiae strain SK1 (MATa/alpha HO gal2 cupS can1R BIO, Kane SM and Roth J. 1974 Bacteriol. 118: 8-14) was cultured in complete medium (YPD, 2% yeast extract, 1% peptone, 2% glucose) for 18 hours, then collected, washed twice with sterile water and resuspended at 108 cells/ml. The cells were biotinylated, labeled and - after treatment of DCs - detected using APC-labeled streptavidin and analyzed by flow cytometry.

These mice were treated with intact anti-CTLA-4 antibodies. This induced the development of regulatory T cells expressing high levels of ICOS and producing IL-10. These regulatory T cells inhibit Th1 responses, in vitro and in vivo, and repress experimental intestinal inflammation, by a mechanism involving IL-10 and IDO.

RCC subjects were deficient in T cell IFN-gamma and IL-2 production pre-vaccination. Their response to DC-based immunotherapy was evaluated.

Lentiviral transduction of shRNAs: to this aim, shRNA specific for IRF4 and STAT3 are in the process to be cloned into the pLVTHM lentiviral vector (obtained from the repository Addgene). This vector has been already successfully used to generate self-inactivating lentiviral vectors previously tested in DCs for transfection efficiency and absence of effects on their phenotype.

In our phase I clinical trial, we highlight the capacity of Dex based-vaccines to restore the number and NKG2D-dependent function of NK cells in 7/14 cancer patients.

We set an in vitro system to study ex-vivo responses able to detect fona fide in vivo primed CD4+ T cells. We investigated spontaneous CD4+ T cell responses to these antigens in normal donors and in patients with high-grade cervical lesions, pancreas adenocarcinoma and advanced melanoma. The antigens were: i) the E6 and E7 proteins of human papilloma viruses, ii) the carcinoembryonic antigen (CEA) and iii) the tumour specific antigen MAGE-A3.