2M KOH were used to dissolve blue formazan particles.

We have chemically coupled the R-epimer of the Pam3CSK4 to long peptides containing a CTL epitope and used this structure to investigate the behaviour of the diastereomer.

We are characterising (in collaboration with Sebastian Amigorena) rat lymph exosomes and will determine if they transport intestinally-delivered scrapie ME7.

The protein vaccines were injected combined with a combination of iNKT cell-agonist alpha-GalCer and MPL, a detoxified version of LPS that signals through TLR4. This combination treatment stimulates antigen-specific T and B cell responses that are greater than those elicited with alpha-GalCer or MPL alone.

The poly(I:C) was used for stimulating DCs. 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. Manufacturer: Invivogen Life Technologies

This PSA was used as a control in the transfection of immature DC. Transfection of immature DCs was done with different constructs encoding for the oncogene Her-2/neu or an adeno virus Her-2 full length construct.

We have prepared two plasmids coding for LACK linked to either eGFP or mCherry fluorescent proteins, under the control of the beta-casein promoter. In order to identify the APCs that were responsible for the development of this state of immune tolerance, we aimed to generate a transgenic mouse that express a recombinant fluorescent protein in the mammary glands, an approach that should result in high levels of expression of the fluorescent protein in the milk. We thus should be able to detect the cells that capture, degrade and present this antigen to T cel...

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.

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

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.

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

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.

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

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.

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.