Co-electroporation of immature DCs with both poly(I:C(12)U) and Melan-A/MART-1 encoding mRNA induced strong anti-Melan-A/MART-1 CD8(+) T-cell responses in vitro. Higher numbers of Melan-A/MART-1-specific CTLs were consistently obtained with poly(I:C(12)U)-activated DCs compared to DCs matured in the presence of an inflammatory cytokine cocktail.

The melanin-free total tumor RNA is obtained from melanoma metastases; it is suitable for in vitro amplification and DC transfection.

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.

We used a MHC II presentation pathway targeting vector originally published by Wang et al. 1999 for the delivery of antigens in both murine and human DC.

An mAb reacting to the immunodominant LACK156-173 peptide of leishmania bound to I-Ad MHC class II molecules was prepared. To this aim, we injected I-Ad/LACK recombinant dimers to TCR transgenic mice which exhibited an increased frequency of LACK-specific T cells. Four out of 600 supernatants, i.e. 2C44, 2F74, 2E60 and 2X8, readily stained LACK156-173-pulsed fibroblasts, but neither unpulsed fibroblasts nor OVA323-339-pulsed cells. BIAcore measurements yielded equilibrium dissociation constants ranging from 1.1 nM for 2C44 mAb to 120 nM for 2F74 mAb.

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.

Melanoma patients were vaccinated with peptides (MAGE-A3, gp100, NA17 and tyrosinase), with and without montanide as adjuvant. No response was observed when vaccinating without the adjuvant. We conclude that Montanide had a clear adjuvant effect for the CD8 T cell responses measured against these four peptides.

This is mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3). It was used to electroporate autologous dendritic cells in order to produce a vaccine for advanced melanoma patients.

These multimers were provided by P. Coulie and used to establish routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry.

The expression and function of these murine FcgammaR in CD11c+CD11b-B220+ plasmacytoid DCs (pDCs) was investigated. pDCs express mostly FcgammaRIIB while the expression of FcgammaRI and FcgammaRIII is only detected by RT-PCR at low but significant level. Moreover, the ITAM-containing intracellular chain associated to FcgammaRI and FcgammaRIII is strongly expressed in pDCs as detected by biochemical assay.

Melanoma patients were vaccinated with 4 peptides (MAGE-A3, gp100, NA17 and tyrosinase) presented by HLA-A2 and monitored. We measured responses to these peptides injected with Montanide, but did not know whether the Montanide adjuvant was important for these T cell responses. We therefore injected the same peptides without adjuvant, and no response was observed. We conclude that Montanide had a clear adjuvant effect for the CD8 T cell responses measured against these four peptides.

1% (vol/vol) non-essential amino acids were added to the monocyte cell culture.

Direct administration of ovalbumin (OVA) encoding lentiviral vectors caused in vivo transduction of cells that were found in draining lymph nodes (LNs) and induced potent anti-OVA cytotoxic T cells similar to those elicited by ex vivo transduced DC.

The vaccine for ovarian carcinoma is composed by autologous DCs pulsed with apoptotic autologous ovarian carcinoma cells. Apoptosis is induced by UV.

This vaccine was used in patients with colorectal cancer or ovarium cancer. It was well tolerated and robust p53-specific T cell responses were induced.

PE-alpha DC80 is used for staining of incubated cells for FACS analysis for activation markers.

50 U/ml of penicillin were added to the monocyte cell culture.

This vaccine consists of G4-DC loaded with 8 different peptides. It was tested in a trial in small cohorts of patients (3-9) who were vaccinated with differently composed DC-vaccines.

This PGE2 was used in a DC maturation process with the final aim to produce a DC vaccine.

The CpG was used for stimulating DC. It was shown in mice that both IL-10 as well as IL-12 production is dependent on the signalling adaptor molecules either MyD88 or TRIF, respectively in response to CpG, LPS or PolyIC.