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Datasets  >  preclinical study dataset  >  Establishment of immunomoni...

Establishment of immunomonitoring vaccination trials with RNA transfected DC by using overlapping peptides covering the whole protein sequence of the tumor antigens / RNAs used

preclinical study dataset

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.





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