Tissue biopsies of the last vaccination site and of the VIN lesion prior to the first vaccination and 3 months after the last vaccination were analysed for the presence of HPV16-specific T cells. The IFN?-ELISPOT analysis and proliferation with its accompanying cytokine production revealed a strong and broad vaccine-induced T cell response. The strength of the immune response (defined as breath and magnitude of the response) was significantly higher in the patients with a complete remission (CR) than in the patients with no changes (NC) in the lesion size. Ph...

Tissue biopsies of the last vaccination site and of the VIN lesion prior to the first vaccination and 3 months after the last vaccination were analysed for the presence of HPV16-specific T cells. The IFN?-ELISPOT analysis and proliferation with its accompanying cytokine production revealed a strong and broad vaccine-induced T cell response. The strength of the immune response (defined as breath and magnitude of the response) was significantly higher in the patients with a complete remission (CR) than in the patients with no changes (NC) in the lesion size. ...

We have used “conventional” techniques to monitor the vaccine induced specific anti-tumor T cell responses (thymidine-incorporation, ELISA, LDA and ELISPOT assays). We have set the staining conditions for the HLA-DR*1101 tetramers loaded with tetanus toxoid and MAGE-3 peptides corresponding promiscuous CD4+ T cell epitopes. Antigen-specific CD4+ T cells are visualized after in vitro short-term expansion; we are currently optimizing the conditions for the ex-vivo staining.

Tissue biopsies of the last vaccination site and of the VIN lesion prior to the first vaccination and 3 months after the last vaccination were analysed for the presence of HPV16-specific T cells. The IFN?-ELISPOT analysis and proliferation with its accompanying cytokine production revealed a strong and broad vaccine-induced T cell response. The strength of the immune response (defined as breath and magnitude of the response) was significantly higher in the patients with a complete remission (CR) than in the patients with no changes (NC) in the lesion size. Ph...

Tissue biopsies of the last vaccination site and of the VIN lesion prior to the first vaccination and 3 months after the last vaccination were analysed for the presence of HPV16-specific T cells. The IFN?-ELISPOT analysis and proliferation with its accompanying cytokine production revealed a strong and broad vaccine-induced T cell response. The strength of the immune response (defined as breath and magnitude of the response) was significantly higher in the patients with a complete remission (CR) than in the patients with no changes (NC) in the lesion size. Ph...

We attempted to improve the adoptive transfer protocol for immunotherapy by stimulating T cells with monocyte derived DC pulsed with tumor lysate, instead of simply adding tumor lysate into PBMC cultures. T cells raised exhibited some tumor reactivity and outgrowth of a distinct T cell population could be observed.

By using TAP deficient cells, we found that TAP is also necessary for the process of presentation of the antigen in MHC class I. We further characterized the antigen depot by confocal microscopy and found that the antigen co-localized with the lysosomal marker LAMP1, but not with the endosomal marker EEA1, TAP, MHC class I or MHC class II. We conclude that the antigen depot is a storage compartment and not a loading compartment. We have started to examine depot formation by targeting antigen to other receptors. For example, we have found that presentation ...

We have determined mechanisms for DC-induced Th1 development and the molecular pathways for inducing Th1 cells producing IFN-gamma solely, or Th1 cells producing IFN-gamma and IL-10, which have important implications for regulation of the immune response to eradicate pathogens with minimum pathology.

These T cells stem from an elutriation process from a metastatic lesion of stage III/IV melanoma patients. Subsequently, these T cells will be expanded by co-culture with tumor lysate loaded DC.

These TcR-Tg clones are specific for the P14-GP33 antigen. In the individual T cells, we studied the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

The TcR-Tg clones are specific for the 0T-1-OVA antigen. In the individual T cells, we studied the expression of twenty different genes either mediating effector functions, or coding for different receptors involved in T cell differentiation and memory generation.

It was found that these cells do not migrate to lymph nodes in the steady state.

For studying in vivo responses to antigens, we initiated a long program comparing the behaviour of TCR-Tg cells specific for GP-33 peptide.

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.

Concerning « 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 have determined mechanisms for DC-induced Th1 development and the molecular pathways for inducing Th1 cells producing IFN-gamma solely, or Th1 cells producing IFN-gamma and IL-10, which have important implications for regulation of the immune response to eradicate pathogens with minimum pathology.

It was evaluated whether STAT-1 was phosphorylated in DC following activation with TLR-dependent Th1 and Th2 stimuli and we found that only in presence of TLR stimuli able to induce Th1 responses STAT-1 was activated.

The D2SC1/Flt3-DC were stained (Mini67-1KT or Mini26-1KT (PKH67/PKH26 green/red fluorescent cell linker mini kit for general cell membrane labelling) from Sigma) for a process of dead cell uptake.

The dead cells were stained (Mini67-1KT or Mini26-1KT (PKH67/PKH26 green/red fluorescent cell linker mini kit for general cell membrane labelling) from Sigma) for a process of dead cell uptake by DC.

These cells were stained with anti-CD11c antibodies for a process of dead cell uptake by DC.