Surface markers were used to establishe routine monitoring of tumor-specific T-cell subsets by multicolor flow cytometry. For this purpose multimers were used kindly provided by Pierre Coulie (Brussels) together with CD3 surface markers (and additionally, CD8, CD4, CD45RA, CCR7, CD25, CD27, CD137, CD127) in different 8-color panels on the BD FACSCanto II. Additionally, functional markers such as CD107a were used and intracellular cytokine staining for the further analysis of individual T-cell populations and T-cell clones.
Establishment of immunomonitoring vaccination trials with RNA transfected DC by using overlapping peptides covering the whole protein sequence of the tumor antigens / RNAs used