Activated T cells are detected by their expression of CD137.

All the individual microcultures are then screened for recognition of autologous EBV-B cells transduced with a retrovirus encoding the protein. Non transduced EBV-B cells are used as control stimulator cells.

PBMC are stimulated in limiting dilution conditions with a pool of overlapping peptides (15 amino acids) covering the protein.

Purified CD8 T cells using MACS cell separation are co-cultured with electroporated DC.

The polarization is assessed by flow cytofluorimetry and cytokine production.

Untouched naive CD4+ T cells are co-coltured with fungi -pulsed DCs

The DC with blocking receptors are exposed to fungi.

The DCs are treated with the receptor antagonist laminarin, with chitin and mannan.

The CD8 T cells become evaluated on their cytokine secretion, cytotoxicity and % of antigen specific T cells.

This step is repeated several times to do several stimulations.

Dendritic cells are electroporated.

CD8 T cells are purified using MACS cell separation.

Advanced melanoma patients are vaccinated with the DC vaccine.

refolded CD1 molecules are used to generate CD1 tetramers

Electroporation of autologous DCs electroporated with mRNA encoding CD40L, CD70, caTLR4 and one tumorantigen (gp100, Tyrosinase, Mage-C2 or Mage-A3) to produce the vaccine

The DC populations were CD123+, CD11c+CD16+, CD11c+BDCA1+, CD11c+BDCA3+ DC.

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

Pulsing of autologous DCs with apoptotic autologous ovarian carcinoma cells.

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

Patients with advanced prostate carcinoma are vaccinated with the DC vaccine.