Establishment of immunomonitoring vaccination trials with RNA transfected DC by using overlapping peptides covering the whole protein sequence of the tumor antigens / RNAs used
In 2007 we established routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry and obtained immunological data from this. For this purpose we used multimers kindly provided by Pierre Coulie (Brussels) together with surface markers (such as CD8, CD4, CD3, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally we used functional markers such as CD107a and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.
Monitoring of the ongoing vaccination trial with RNA transfected autologous DC was done with overlapping peptides covering the whole protein sequence of the tumor antigens MageA3, MelanA and Survivin. With this approach we monitored the immune responses of 8 vaccinated patients and of more than 15 other stage IV melanoma patients. Results showed broad preexisting T-cell responses in some patients and de novo immune reactivity in 3 vaccinated patients. The fine specificity of these immune responses needs to be further defined.
Demonstration that defined re-stimulation Elispot correlates very well with MPLC assay.
data set
preclinical study dataset
CD8
CD4
CD3 receptor
CD45
CD25 receptor
CD137 receptor
human CD127 receptor
Melan A
melanoma antigen MAGE-3
survivin
CD107a receptor
CD27 receptor
CD197
ribonucleic acids
peptide
tumor associated antigen
T cell
autologous dendritic cell
Homo sapiens
BD FACSCanto II
MPLC assay
Multicolor flow cytometer
Overlapping peptide library technique for monitoring RNA-DC vaccinated patients
Elispot analysis
Alexander Steinkasserer
Gerold Schuler
CCR7 surface markers
CD107a functional markers
CD127 surface markers
CD137 surface markers
CD25 surface markers
CD27 surface markers
CD3 surface markers
CD4 surface markers
CD45RA surface markers
CD8 surface markers
Melanoma patients
Multimers
RNA transfected autologous DC
cellular modification design: RNA interfering experiment