Our group has MRI data available on the migration of DCs in vivo in melanoma patients. From isolated lymphnodes obtained after surgery we could identify single DC after staining with Prussian blue for iron. We are in the process of analysing the T cell rosettes (activation stage) that surround these DC to get insight in the functional activity. It was also demonstrated that DCs transfected with RNA produce derived proteins in vivo in the lymph nodes. Also efforts were spent to get further information on the dynamics of immature and mature DCs. We discovered (van...

Our group has obtained proteomic data on the three known splenic subsets (CD8+, CD4+, DN DC). Despite the significant advances in mass spectrometry, which has enabled much of proteomics, due to various analytical challenges so far no eukaryotic total cell proteome has been sequenced completely. An alternative approach is to purify single organelles or DC components in order to generate the proteomic data. An additional problem is to obtain highly pure fractions of DC from crude population of untouched splenic cells with a purity of 70-80%. This includes several...

Data is available on human Hemato-Lymphoid System Rag2gc-/- mice infected with both CXCR4 as well as CCR5 tropic HIV-1 strains. HIV causes a disseminated infection and spreads in all newly generated lymphoid tissues, thus closely resembling HIV infection in humans. However, as in EBV infected animals, adaptive T and B cell responses were not consistently detected. Thus, although these “humanized” mice represent a major step forward in generating an in vivo preclinical testing system, they still harbor limitations as inconsistent/weak adaptive immune respons...

Our lab has immunological data available on human Hemato-Lymphoid System Rag2-/-gc-/- mice infected with EBV. Both Epstein Bar Virus (EBV) and Human Immunodeficiency Virus (HIV) are human specific lymphotropic viruses. Human Hemato-Lymphoid System Rag2-/-gc-/- mice were infected with EBV and mount an immune response (i.e. cytotoxic T cell proliferation, some control of EBV driven B cell proliferation, perforine and granzyme expressing T cell infiltration in B cell infected areas in lymphoid organs in situ), however, specific T cells could not be detected directl...

We have data available on some functional immune responses in a small animal model to study the development and function of the human hematopoietic and immune system as well as human specific infections in vivo. We established “human hemato-lymphoid-system mice” by transplanting human CD34+ cord blood cells into irradiated newborn Rag2-/-gc-/- mice, leading to de novo development of human B, T, and dendritic cells; formation of structured primary and secondary lymphoid organs; and production of some limited functional immune responses.

We have data available on the role of chemokine receptors in regulating migration of mature DC and subsets of effector and memory T cells in secondary lymphoid organs. CCR7 is a chemokine receptor that is expressed by mature DCs and central memory T cells (TCM) and regulate migration of these cells in secondary lymphoid organs in the steady state. T lymphocytes lacking the lymph node homing receptors L-selectin and CCR7 do not migrate to lymph nodes in the steady state. In contrast it was found that lymph nodes that drain sites of mature DC or adjuvant inoculatio...

We have continued recruitment of patients and obtained first data in the two clinical trials of active vaccination in metastatic melanoma. Structural works that have been performed to up-grade the GMP facility for conduction of phase III clinical trials has limited the number of patients recruited. Nonetheless, we were able to complete the vaccination schedule in 4 patients and to perform additional injections in disease free patients, in patients with stable disease or with slowly progressive disease. We could not recruit new patients because the generation in GM...

Our lab obtained data on functional homogeneity from a gene expression analysis at single-cell level for the expression of 8 effector genes from normal donors. Until 2006 we subdivided CD8 PBL from normal donors in 14 different cell sets based in the expression levels and the co-expression of 5 different cell surface markers and studied functional homogeneity by screening cells from each subset at single-cell level for the expression of 8 effector genes (Blood, 2007). Although these studies allowed us isolate some homogeneous cell types, we yet found heterogenei...

Our lab obtained data on the kinetics of effector gene expression and association in TCR-Tg CD8 cells responding to the male antigen and to Listeria-OVA with regard to « in vivo » responses to Ags. Surprisingly, instead of a single program of differentiation leading to the coordinated expression of all effector genes; we found that individual effector genes had different kinetics of expression and associated randomly generating several new effector cell types. These results are published in J. Exp. Med. Since these results were obtained in TCR-Tg mice, and thus ...

We obtained data from our activity to establish the identification of versatile central memory CD8+ T-cells capable of high IL-2 secretion according to new markers. Versatile human CD8+ T-cells represent up to 30% of the CD8+ central memory pool in healthy adults and are characterised by unique functional capabilities (ability to secrete IL-2 after restimulation in concordance with low effector cytokine (e.g. IFN) and lack of cytotoxic mediator (e.g. perforin) expression (Frentsch et al., unpublished)). They might present an attractive candidate memory/effector C...

We obtained data from an in vitro system to study ex-vivo responses able to detect bona fide in vivo primed CD4+ T cells. We focused on the monitoring of tumour antigen specific CD4+ T cell responses. We select the following tumour associated antigens: i) the E6 and E7 proteins of human papilloma viruses, ii) the carcinoembryonic antigen (CEA) and iii) the tumour specific antigen MAGE-A3. 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 ...

We obtained transcriptional and pathway analysis data of human monocyte-derived DCs stimulated with yeast, spheroplasts, pseudohyphae, spores, purified RNA and not purified RNA and LPS. We propose the role of cell wall in the mechanisms of DC-Pathogen interaction as an experiment that could serve as reference in bioinformatics approaches for the reconstruction of DC activation signalling pathways. We have stimulated human Monocyte derived Dendritic Cells (hMoDC) with yeast, spheroplasts, pseudohyphae, spores, purified RNA and not purified RNA, LPS. We have measur...

Data is available from a re-analysis of existing microarray data; we started experimental work to assess the reliability and completeness of the DC_Eu.Gene pathway set using stimulation of DCs with S.cerevisiae cells, pseudohyphae, spores in comparison with the results obtained with LPS, R848, LPS and R848, Zymosan and Curdlan.

We annotated DC specific pathways and integrated them in the pathway tool. Initially we have annotated DC specific pathways using information publically available on the web. A preliminary curated non exhaustive pathway list can be found in the DC-THERA version of the program DC_Eu.Gene.

Our lab generated integrated microarray data on cellular pathways and gene ontology annotation. This has been done in the context of the program DC_Eu.Gene freely available for the DC-THERA users, using ENSEMBL as a conversion table.

Data has been obtained from the application of a DC-based assay aimed at identifying new adjuvant molecules potentially capable to induce Th1 responses, determining the effects of new chemical compounds on dendritic cell capacity to produce type I IFNs. The selected molecules could be potential new vaccine adjuvants.

Preclinical data is available on the power of preventing disease and invoking immune response of our lead product for the prevention of infectious disease, ImmunoVexHSV2, a vaccine for genital herpes. ImmunoVex HSV2 is a novel live-attenuated vaccine candidate that expresses approximately 80 HSV-2 proteins intended to stimulate a broad and powerful immune response. In preclinical studies, ImmunoVex HSV2 completely prevents disease and invokes a powerful immune response. We intend to initiate a Phase I clinical trial for this product early in 2008.

Data has been obtained from our phase 1 HIV clinical study described here: Study Design: Patients that are successfully treated with highly active anti-retroviral therapy (HAART) with no measurable viral load AND have a cryopreserved infectious plasma sample that was drawn immediately prior to initiation of HAART are administered 4 monthly doses of the Arcelis product. Current status: This trial is fully enrolled. Immune response data is encouraging in the first 7 patients for which the analysis has been completed. Six of 7 patients developed CD8+ T-cell immune...

Data have been obtained from a phase 2 study testing PME-CD40L DC in RCC patients with clear cell histology. The Phase 2 trial has the same general design, dosing regimen, endpoints, and immunomonitoring plan as the completed RCC trial. However, the Phase 2 study tests the improved RCC product (i.e., PME-CD40L DC) and is restricted to RCC patients with clear cell histology. As predicted from the in vitro data, the improved PME-CD40L DC product does indeed lead to restoration of both IL-2 and IFN-? responses. Also consistent with our in vitro observation is the...

First clinical vaccination data has been obtained using an HSV vector expressing full length tyrosinase, gp100 and MART-1 to transduce DC from melanoma patients. Biovex has developed HSV-based vectors optimised for antigen delivery to dendritic cells by the deletion of HSV genes which usually inhibit DC function. These deliver antigens to DC at very high efficiency and activate the transduced cells similarly to LPS. Based on this technology, Biovex began a clinical program where an HSV vector expressing full length tyrosinase, gp100 and MART-1 was used to transdu...