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...

We have data available on a novel M-CSF dependent DC developmental pathway that is independent of Flt3L. These DC have unique characteristics and precursor origin. The cells can be found in vivo in Flt3L gene deleted (-/-) mice injected with recombinant M-CSF.

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...

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 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...

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.

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.

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...

We have data from a clinical trial with melanoma patients (stage IV, documented progress) receive DC transfected with RNA encoding MelanA, Mage3 and Survivin +/- E/L-selectin by the i.v. route. This DERMA-ER-DC 05 trial (multipeptide loaded cytokine matured moDC + prior Treg elimination by 3x ONTAK treatment [5µg/kg], 8 evaluated pat.) did not show any enhanced immune reponses compared to the equivalent cohort in the DERMA-ER-DC 04 trial. Patient recruitment into the 2nd phase of the DERMA-ER-DC 06 trial has started (sequential adaptive design). Immunomonitori...

We developed additional antigen-loading strategies for DC and obtained data from first experiments on which constructs lead to antigen presentation and which do not. On the one hand we compared lipofection of tumor-antigen-encoding RNA to RNA electroporation into DC, and found that electroporation results in antigen expression in all cells, while lipofection results in higher antigen expression only in a portion of the cells (other cells are negative). Interestingly, the priming capacity of MelanA/HLA-A2-specific autologous CD8+ T cells by lipofected DC was high...

Data is available on the interaction between microorganism-associated molecular patterns and individual toll-like receptors on dendritic cells that turn on signal transduction pathways leading to the activation of different transcription factors. Microbial invasions are perceived by pattern recognition receptor (PRR)-expressing cells of the innate immune system. PRRs bind a number of microbial products collectively referred to as microorganism-associated molecular patterns (MAMPs). Toll-like receptors (TLRs) are the best-characterized PRRs. The interaction of MAM...

Our lab has data available on the effect of endogenous IL-10 production by DC subsets resulting in low levels of IL-12p70 production in mice. We have shown that plasmacytoid pDC respond to both CpG and the TLR-7 ligand R848 to produce very high levels of IL-12p70 (in the ng/ml range per 5 x 105 cells/ml) and IFN-gamma. However, BM-myeloid and splenic CD8alpha+ DC produced at least fivefold less IL-12p70, and CD8alpha- DC produced undetectable levels of IL-12p70 upon stimulation with CpG. We show that induction of endogenous IL-10 production by these latter DC ...

We can provide one experiment with 32 micorarrays, two experiments with 8 microarrays and one experiment with 12 micorarrays. The yeast oligonucleotide array was constructed using the S. cerevisiae Genome Oligo Set ™ (Operon Technologies, CA, USA) composed of 6240 optimized oligonucleotides (70mers) each representing one yeast gene.