Differentiation of monocytes into dendritic cells is promoted, along with recombinant IL-4, by addition of granulocyte-macrophage colony-stimulating factor (GMCSF 1000U/ml, Chemicon).

TNF-alpha is used during maturation of the DCs.

Differentiation of monocytes into dendritic cells is promoted, along with granulocyte-macrophage colony-stimulating factor, by addition of recombinant IL-4 (1000U/ml, R&D Systems).

Constructs for fusion proteins of the DC specific receptor DC-SIGN fused to GFP have been completed and tested.

We analysed the ability of invariant NKT (iNKT) cells to assist priming of antigen specific T and B cell responses. 1) We have demonstrated that activation of human DC by Toll like receptor ligands (TLR-L) modulates the lipid biosynthetic pathway, resulting in enhanced recognition of CD1d-associated lipids by iNKT cells. 2) We have clarified the mechanisms by which CD1d-restricted lymphocytes translate T cell receptor (TCR) recognition of lipids with similar group heads into distinct biological responses. Using a soluble iNKT TCR and a newly engineered anti...

Fabs that bind strongly to L-SIGN, but to a lesser degree or not at all to dendritic cell-specific ICAM-grabbing nonintegrin (DC-SIGN) were isolated and characterized. We believe that the isolated Abs may be useful for selective delivery of Ags to DC-SIGN- or L-SIGN-bearing APCs for the modulation of immune responses and for blocking viral infections.

Tetanus toxide epitope embedded into a L-SIGN/DC-SIGN-cross-reactive Ab was targeted to dendritic cells. We believe that the isolated Abs may be useful for selective delivery of Ags to DC-SIGN- or L-SIGN-bearing APCs for the modulation of immune responses and for blocking viral infections.

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.

HLA-DP4/MAGE3 molecules have been prepared to stain ex vivo PBL from immunized melanoma patients for a study of anti-tumor responses in melanoma patients that were vaccinated with autologous DCs loaded with a combination of tumor Ag peptides.

The peptides are used in the generation of monocyte-derived DC (pulsing of the cells at a concentration of 5 µg/mL in approximately 5 mL and at a cell density of 20 x 10e6/mL for 1 h.).

Patients with resected HPV16-positive cervical cancer were vaccinated with an overlapping set of long peptides comprising the sequences of the HPV16 E6 and E7 oncoproteins emulsified in Montanide ISA-51.

Patients with resected HPV16-positive cervical cancer were vaccinated with an overlapping set of long peptides comprising the sequences of the HPV16 E6 and E7 oncoproteins emulsified in Montanide ISA-51.

We have generated a monoclonal antibody, 2C44, that reacted to a peptide derived from the Leishmania LACK protein bound to I-Ad MHC class II molecules. We have successfully used the 2C44 mAb to visualize LACK-presenting DCs in mice infected with the L. major.

We showed that, in the absence of additional stimuli, inflammatory signals such as pro-inflammatory stroma-derived cytokines on DC are ineffectual in promoting DC activation and cannot substitute for engagement of innate receptors directly on DC.

The model antigen OVA was fused to the C1C2 exosome-binding domain of MFG-E8/lactadherin. This construct was used in a study on the capture of antigen-bearing exosomes by DCs and cross-presentation of tumour antigen in exosomes.

It has been shown that DCs express specific receptors for exosomes. Since ICAM-1 on exosomes is required for their ability to activate T cells via DCs in vitro, the role of Mac-1 and LFA-1 (the major ligands for ICAM-1) as potential receptors for exosomes has been investigated. A new role for LFA-1 on DCs, as a receptor for exosomes has been proposed.