Flow cytometric analysis data were obtained of translation activation in DC-activated T cells.

We have obtained data by fluorescence microscopy analysing DC distribution in spleen and how this is affected during inflammation driven by stromal cells or hematopoietic cells. One of our images of DC was selected as the cover for the June 2007 issue of Nature Immunology.

We obtained data from studies of CD8 T cell properties in man comparing HIV, CMV and EBV-specific cells. It is usually assumed that effector gene expression in CD8 T cells may be similar in all immune responses, CD8 T cells in vivo differentiating into cytotoxic T cells that also produce g-inf. In contrast we published in 2007 that CD8 follow several differentiation waves and that effector gene expression is heterogeneous, different effector genes having different kinetics of expression-down regulation and associating randomly, suggesting that different immune re...

Our lab has gathered data in establishing a per-clinical model of breast cancer.

Knowledge and gene expression data were gained on differentiation patterns of T cells.

In 2007 we established routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry and obtained immunological data from this. For this purpose we used multimers kindly provided by Pierre Coulie (Brussels) together with surface markers (such as CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally we used functional markers such as CD107a and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones. Monitoring of the ongoing vacc...

We investigated the effects of tumour hypoxia on DC differentiation and maturation and gathered data in mice. This is a an important topic as sites of inflammation are often characterized by both low oxygen tension and an extensive infiltration of inflammatory leukocytes, including DC, which need to move against oxygen gradients. We showed that oxygen availability is necessary to promote full monocyte-derived DC differentiation and maturation. Low oxygen tension (hypoxia) inhibits expression of several differentiation and maturation markers (CD1a, CD40, CD80, CD8...

Data were produced in our lab on a broad spectrum of priming events and their impact on the effector and memory phases of the CTL response. Activation of a cytotoxic T lymphocyte (CTL) response in an antigen-exposed lymph node involves a great diversity of encounters between naive CTLs and APCs that differ in both duration and quality. This broad spectrum of priming events instigates a complex blend of CTL developmental pathways. Using an experimental system that allows tight control over CTL priming, we have singled out defined priming events and analyzed the ...

We have generated data on the developmental control of translation during DC activation. Our work (Lelouard et al., JCB) demonstrates that DCs down-regulate and change the quality of translation in order to increase their survival, however during this process MHC I antigen processing is affected. This has important consequences for the way antigens should be introduced in DCs and how long after stimulation they should be used during vaccination application. We have also initiated studies on the impact of dsRNA on the regulation of translation and the functional re...

Design of this trial: sequential adaptive design, 2nd cohort, mature DC transfected with RNA encoding MelanA, Mage3 and Survivin +/- E/L-selectin, i.v. route, 13 evaluated patients. Comparison of pre-existing and vaccine-induced immune responses in stage IV melanoma patients. We have efficiently transfected DC with RNA encoding a functional protein (E/L-selectin), which allows entry of DC into LN from HEV. These DC rolled in vitro on sialyl-LewisX-coated slides, and in vivo, mouse E/L-selectin-transfected DC homed to LN after i.v. application. Monocyte-derived and...

We have data on a novel innate signalling pathway in DC. Using curdlan as a specific agonist of dectin-1, we have shown that this C-type lectin couple to Syk kinase leads to activation of ERK, JNK and p38 MAPKs, as well as NF-kappaB. However, signalling terminates rapidly once the receptor is internalised, thereby explaining previous observations that large, non-phagocytisable Dectin-1 ligands are much more potent agonists than smaller ones. This is the first example of a bona fide signalling pathway from a cell surface innate receptor other than a TLR. We will ...

Our group obtained data on molecular signaling triggered by LPS in dendritic cells and the role of CD14 in the regulation of the DC life cycle through NFAT activation. Toll-like receptors (TLRs) are the best-characterized pattern recognition receptors (PRRs). Individual TLRs recruit diverse combinations of adapter proteins, and thereby trigger signal transduction pathways leading to the activation of various transcription factors, including nuclear factor (NF)-kB, activation protein (AP)-1, and interferon regulatory factors (IRFs). Interleukin-2 (IL-2) is one of th...

Our lab has data from an interim statistical analysis of the DERMA-ER-DC 04 trial (multipeptide loaded cytokine matured moDC +/- CD40L activation, 70 evaluated patients). This analysis for the first time showed a clinical correlation of induced immune responses as measured in the blood with outcome, as there have been so far statistically significantly less events in stage III patients showing good CTL reponses (IFN-y Elispot) to multiple class I and II peptides compared to low responders of the stage III cohort. Additional in vitro maturation of DC by CD40L (leuci...

We have obtained data on a possible cooperation between myeloid and plasmacytoid DCs in response to different microbial stimuli and a characterization of the mechanisms of this cooperation. For this, we have studied co-culture of mDCs and pDCs in response to CpGs, LPS and bacterial particles. Taking advantage on the fact that human myeloid and plasmacitoid DC show a selective response to LPS and CpG respectively, we have previously identified a cross-talk between two type of cells. Indeed it was observed that LPS-stimulated mDCs are able to induce up-regulation o...

Data has been obtained on the effect of exposure of DCs or gamma-delta T lymphocytes to HIV-1 and on the miR profile expressed in HIV-exposed DCs. Based on our previous findings demonstrating a bidirectional activating interaction between immature MDDCs and gamma-delta T lymphocytes (Conti et al., JI. 2005), as well as on the demonstration that HIV-1 exposed MDDCs exhibit an impaired functional maturation (Fantuzzi et al., J Virol 2004), we are currently investigating whether exposure of DCs or gamma-delta T lymphocytes to HIV-1 can directly modulate their functi...

Pathway-level data have been obtained from a pilot experiment on yeast cells treated with peptides with potential antifungal action. We have compared results from both genomics and proteomics experiments and search for similarities at the level of pathways and have used Eu.Gene to compare the two different data sources demonstrating the feasibility of using this program to this task. Overall correlation between changes in RNA and protein expression was poor but increased when looking at components of immunologically-relevant pathways and Gene ontology terms. The...

Understanding the distribution, function, and lineage relationship of CD8+ T-cell subpopulations is of fundamental value for the monitoring of the immune system in several experimental and clinical situations. However, the available data concerning the description of effector and memory CD8+ subsets in humans remain rather fragmentary because different studies favored the usage of distinct and restricted sets of cell surface markers and functional parameters. We obtained coexpression data of 18 genes simultaneously in individual cells from CD8+ T cell subsets (as...

We are planning clinical trials of melanoma patients treated with DC and tumour lysate plus adoptive transfer of T cells which will result in corresponding data.

We have obtained imaging data by confocal microscopy of translation activation in DC-activated T cells and have pursued our efforts on the definition of a novel way of monitoring DC and T cell activation as well as studying viral infections. We also have now applied the puromycin-based technology developed in our laboratory (SUnSET, surface sensing of translation) to monitor translation by FACs in individual or cell populations. We have demonstrated that translation activation is an excellent read-out for early antigen specific T cell activation by DCs. We hope tha...

Gene expression data were generated on DC activated with different TLR agonists typically associated with the respective response.