We have developed immunological techniques for monitoring of tumour antigen specific CD4+ T cell responses. We set an in vitro system able to detect fona fide in vivo primed CD4+ T cells. Purified CD4+ T cells are first stimulated in vitro once in the presence peptides, previously identified as containing naturally processed epitopes on tumour-associated antigens. After two weeks, proliferation and cytokine secretion (IFN-?, IL-5, IL-4, IL-2, TNF-?, IL-10, TGF-?, IL-17, granzyme and perforin) are tested. To estimate the frequency of antigen specific CD4+ T cell we us...

This puromycin-based technology was developed in our laboratory to monitor translation by FACs in individual or cell populations. It is a non-invasive method to monitor protein synthesis and cellular activation in single cells or heterologous populations.