From the CD14- PBMCs, untouched naive CD4+ T cells were isolated with a combination of magnetic sorting.

The expression and function of murine FcgammaR in these CD11c+CD11b-B220+ plasmacytoid DCs was investigated.

We have demonstrated that the target of FcgammaRs with the antigen induces the internalization of the immune complex, leading to the maturation of CD11c+CD11b+ conventional DCs. These DCs become thus efficient to present antigenic peptides to helper CD4 T cells and cytotoxic CD8 T cells via cross-presentation pathway.

To quantify presentation of the epitopes by DC, we used bulk T cells electroporated with TCR-encoding RNA in stimulation assays.

These bone marrow derived murine dendritic cells (BM-DC) were generated with GM-CSF according to Lutz et al. (J Immunol Methods 1999, 223: 77-92) and were used for the investigation of chemokine dependent CCR7 signalling.

The properties of this DC population and its production of cytokines in response to different Toll-like receptor (TLR) agonists were evaluated. The TLR agonists used were: PAM3CSK4, Poly I:C, LPS, Flagellin, Imiquimod, Resiquimod, CpG 2216, CpG 2006. BDCA-3+ cells seem not have any capability to respond to TLR agonists.

These autologous DC were used in a phase IB/II study of immunotherapy. Patients were randomized to receive immunizations either with I3 DC (DC generated with interferon-beta and interleukin-3) or with G4 DC (DC generated with GM-CSF and IL-4).

These autologous DC were electroporated with mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3) to produce a vaccine for immunotherapy of advanced melanoma patients.

B cell proliferation (polyclonal, oligoclonal, monoclonal) was observed in human Hemato-Lymphoid System Rag2-/-gc-/- mice after being infected with EBV and mounting an immune response.

The autologous T lymphocytes were stimulated with (frozen) MoDC at a ratio of 1 DC per 10 T cells.

These T cells were generated from the Leukapheresis product, after elutration, of a metastatic lesion of stage III/IV melanoma patients, expanded by co-culture with tumor lysate loaded DC.

These autologous dendritic cells were used for vaccine production. They were pulsed with apoptotic autologous ovarian carcinoma cells.

The autologous DC loaded with KLH, as immunological tracer, and an allogeneic peptidome (i.e. natural tumour peptides, NTPs), obtained from melanoma cell line SK-Mel24, are used for vaccination.

The T blasts are used to immunize MAGE-3 positive patients.

The B16TRex cell line cells were transfected with Bims to induce apoptosis.

B16TRex cell line cells were transfected with Fadd-dd to induce necrosis.

These cells were used to produce a vaccine for prostate carcinoma and were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap cells.

We evaluated the capacity of autologous DC and HEK293 cells transfected with relevant HLA alleles to function as T cell targets in Elispot assays upon transfection with tumor antigen encoding plasmids. Due to strong background from the autologous DC we decided that HEK293 transfected with one HLA-allele at a time plus simultaneously transfected with up to 5 tumor antiges would be optimal to screen for antigen specificity in the patients T cells. However, during a second T cell culture to expand large numbers of T cells for this screening procedure a drastic ex...

These carcinoma cells were used to pulse autologous DCs for vaccine production for ovarian carcinoma patients.

Autologous DC were pulsed with apoptotic allogenic prostate carcinoma cell line LNCap to produce the advanced prostate carcinoma vaccine.