In these patients, dendritic cells were tracked for monitoring of cellular therapy by MRI. MRI cell tracking using iron oxides appears clinically safe and well suited to monitor cellular therapy in humans.

Anti-tumor responses were followed in melanoma patients that were vaccinated with autologous DCs loaded with a combination of tumor Ag peptides.

Melanoma patients were administered ALVAC canarypox virus containing a MAGE-A3 minigene, while others were vaccinated with MAGE-A3 peptide presented by HLA-A1, administered as peptide, and peptide-pulsed DCs. There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

Melanoma patients were vaccinated with peptide-pulsed DC (MAGE-A3, gp100, NA17 and tyrosinase), while other patients were vaccinated with peptides +/s adjuvant or viral vectors (ALVAC canarypox virus containing a MAGE-A3 minigene). There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

The patients were vaccinated with 4 peptides (MAGE-A3, gp100, NA17 and tyrosinase) presented by HLA-A2. We measured responses to these peptides injected with Montanide, but did not know whether the Montanide adjuvant was important for these T cell responses. Indeed, melanoma patients may mount spontaneous responses to these antigenic peptides. We therefore injected the same peptides without adjuvant, and no response was observed. We conclude that Montanide had a clear adjuvant effect for the CD8 T cell responses measured against these four peptides.

Some melanoma patients were vaccinated with the MAGE-A3, gp100, NA17 and tyrosinase peptides presented by HLA-A2 plus the montanide adjuvant, while others were vaccinated with the peptides without the adjuvant or with ALVAC canarypox virus (containing a MAGE-A3 minigene) viral vectors. There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

The melanoma patients were vaccinated with a MAGE-A3 peptide presented by HLA-A1 without adjuvant, while other melanoma patients were vaccinated with ALVAC canarypox virus containing a MAGE-A3 minigene, or peptide-pulsed dendritic cells (DC). There was a correlation between tumor regression and detection of anti-MAGE-3.A1 CTL responses.

It was found that lymph nodes that drain sites of mature DC or adjuvant inoculation recruited memory CD8+ T cells.

These patients are involved in a phase II clinical trial for melanoma and are treated with OncoVexGMCSF. Data so far indicates that OncoVex can destroy both injected and un-injected melanoma deposits. The trial is still in progress.

Patients underwent surgical removal of a metastatic lesion and leukapheresis at the beginning of the trial to provide sources for tumor antigens as well as monocytes and T cells, which will be isolated from the leukapheresis product by elutriation.

4 metastatic melanoma patients were actively vaccinated and additional injections were performed in disease free patients, in patients with stable disease or with slowly progressive disease.

Soluble recombinant HLADR1101 MHC class II molecules receptive for loading with either pathogen or tumor-derived peptides were constructed for a study of CD4+ T cell responses against pathogens or tumors in humans.

We used a MHC II presentation pathway targeting vector originally published by Wang et al. 1999 for the delivery of antigens in both murine and human DC.

Using pharmacological inhibitors, or macrophages and DC from mice carrying a null mutation in MAP kinase signalling molecule(s), we have evidence for a role of certain MAP kinases in the regulation of IL-10 and IL-12, IFN-gamma production.

We have immunized mice with human DC derived-exosomes and generated hybridomas that produce antibodies recognizing at least the human exosomes.

The effect of environmental factors on development of high affinity CTL was investigated using a system whereby OT-1 cells were primed in vitro by engineered APC. After a 20 h priming phase, CTL were transferred to recipient mice that were either naïve, or had been injected with activated dendritic cells one day earlier, thus creating a reactive lymph node.

We showed that, in the absence of adjuvants, DEX mediate potent antigen dependent-antitumor effects against preestablished tumors in mice pretreated with immunopotentiating dosing of cyclophosphamide (CTX).

Subcutaneous injection of hypoxic DCs into the footpads of mice results in defective DC homing to draining lymph nodes, but enhanced leukocyte recruitment at the site of injection.

We infected mice with CD8+ T-cells primed with or without the influence of a reactive lymph node with a flu-virus encoding the specific antigen. After several days we determined the viral load in the lungs with RT-PCR. The results show that in mice with CD8+ T-cells primed with a reactive lymph node the viral load in the lungs was significantly smaller then in a control mice.

Monocytes were cultured in serum free medium in the presence of IL-4 and GM-CSF, and concentrated cytokines were added.