We can provide data for two experiments composed of 14 microarrays each. To obtain the data, DCs were stimulated with different form of fungi. We perform RNA preparation, labelling, hybridization, and scanning according to the Affymetrix reference protocols and BioPolo (Prof. Ricciardi-Castagnoli and Dott. Francesca Zolezzi) and preliminary data normalization according to AMDA (BioPolo).

We can provide Illumina microarray data for four experiments composed of 4 microarrays each on DCs stimulated with pathogenic and non pathogenic fungi. RNA preparation, labeling, hybridization on a HT12 array (Illumina), and scanning were performed according to Illumina instructions by Genomics Lab, Wellcome Trust Centre for Human Genetics (Univeristy of Oxford).

Studies have continued and data obtained on the effects of biolistic (Gene Gun) delivery of vectors engineered to express distinct chemokines at the local skin site of mice. In particular, attention has been focussed on the recruitment of distinct subsets of DC to skin. This work, which is being performed in collaboration with PowderMed, a former SME now owned by Pfizer, may lead to the development of more targeted strategies to induce distinct T cell responses using this technology. A publication has resulted from this work.

Data were obtained on the effect of enhanced CD83 expression on dendritic cells and T cells and its correlation with effective immune responses. Human CD83 is a marker molecule for mature dendritic cells (DC) and is also expressed on activated B and T cells. Although CD83 has been implicated in immune responses, its function on DC and T cells remains unclear. In this study, we wanted to assess the role of CD83 expressed on DC and T cells in the immune response. Down-regulation of CD83 expression on human DC through RNA interference (RNAi) results in a less potent...

We will start a collaboration with G. Schuler (P06) to examine the modulation of dendritic cell function by Denileukin Difitox (ONTAK) and will provide pathway analysis data.

Data was obtained of the capacity of splenic DC, pulsed in vitro with protein antigens or peptides, to induce immunity after transfer into syngeneic animals. To improve existing strategies, the amplitude and polarisation of T cell responses was monitored in the presence or absence of regulatory T cells. Natural regulatory T cells (CD4+ CD25+) were depleted by injection of anti-CD25 mAbs, whereas regulatory T cells were induced by injection of (presumably agonistic) anti-CTLA-4 mAbs. Our observations demonstrate that natural and CTLA-4 induced regulatory T cells a...

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 obtained data from monitoring vaccine induced T cell responses for two clinical trials. We found CD8+ and CD4+ T cell responses against the immunological tracers KLH and TK, and against the NTPs and MAGE-3, respectively. Assays used have been 3H-thymidine-incorporation, ELISA, CBA, LDA and 51Cr-release assays. Autologous tumour specific CD8+ and CD4+ T cell clones have also been obtained from the blood and tumour infiltrating lymphocytes. We investigated the role of spontaneous CD4+ T cell responses against the HPV-18 and 16 E6 and E7 proteins in women wi...

There is data available on translation during DC/T cell interaction in presence or absence of cognate antigenic peptides. Efforts on the definition of a novel way of monitoring DC and T cell activation as well as studying viral infections were finalized. The puromycin-based technology (SUnSET, surface sensing of translation) was applied to monitor translation by FACs during DC/T cell interaction in presence or absence of cognate antigenic peptides. It was demonstrated that translation activation is an excellent read-out for early antigen specific T cell activati...

The initiation of a multi partner trial is planned and data will be generated. The DC-THERA trial will compare the use of ‘TRI-MIX’ RNA in clinical trials that were already underway at each of the three centres (Nijmegen Medical School, Nijmegen; Friedrich-Alexander University, Erlangen; Vrije Universiteit Brussel, Brussels). It was decided that Friedrich-Alexander University, Erlangen, would apply for a licence to prepare GMP-grade ‘TRI-MIX’ RNA and subsequently supply this for all three centres. The multi-centre trial is a well-standardised, two-arm, m...

Multi-photon imaging data on DC-CD8+ T cell interactions in intact lymph nodes (LNs) have been obtained. The initiation of cytotoxic immune responses requires the direct interaction between naïve CD8+ T lymphocytes and dendritic cells. Multi-photon imaging showed that during priming, naïve T cells and dendritic cells (DCs) establish sequentially brief (i.e. minutes) and long (hours) antigen-specific contacts. We observed that ICAM-1 expression by mature DCs is critical for long-lasting contacts with CD8+ T cells, but dispensable for short-lived antigen-specific i...

Our group has produced in-vivo multi-photon microscopy imaging data on the distribution and motility of NK cells by tracking migration of DC-primed NK cells in vivo. Since LPS-activated DC are potent inducers of NK cell priming and lymph nodes appear to be a key place in which DC and NK cell interactions occur, we have investigated the in vivo capacity of resting and DC-primed NK cells to reach the draining lymph nodes. DC –activated NK cells reach the draining lymph node within 24 hours after intravenous injection and persist there for at least 72 hours. NK cell...

Data has been generated by multiplex analysis on the response of human MoDC to yeast, spheroplasts, pseudohyphae and spores. Human blood contains myeloid (mDCs) and plasmacytoid (pDCs) dendritic cells that differ in cytokine secretion pattern and responsiveness to microbial stimuli. We found that LPS-stimulated mDCs are able to induce up-regulation of co-stimulatory molecules on co-cultured pDCs. Likewise, CpG-stimulated pDCs activate co-cultured mDCs. The cross talk between these two populations of DC is very efficient since it can be observed at mDC/pDC ratio ran...

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.

Our lab has obtained data on maturation stimuli for GMP-grade DC.

Pathways-based analysis will be done in collaboration with P15 (Cavalieri) to identify the intracellular target of Japanin and data will become available from this study.

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 was obtained on the efficacy in a phase I/II study investigating multiple injections of CLL-DCV01 in patients with previously treated B-cell chronic lymphocytic leukemia. This study is fully enrolled (n=9). Efficacy data will be assessed via tumor burden data collected over the next two months. Absolute lymphocyte counts and immunomonitoring data will be compiled by the end of Q1’08.

The study protocol that will be used for this trial has been reviewed by the local ethical committee and is now ready to be submitted to Läkemedelsverket, the Swedish regulatory authority for clinical trials. Upon approval by Swedish authorities we plan to include, treat and evaluate 10 patients during a period of one to two years. It will probably start in 2009 and data will become available.

We have obtained data from a phase I long peptide vaccination trial with p53 peptides in Montanide adjuvant in patients with colorectal cancer or ovarium cancer. This vaccine was well tolerated and robust p53-specific T cell responses were induced. In the future we intend to combine the p53 vaccine with other cancer therapies such as chemotherapy.