Infusions of T cells engineered with TCRs directed against tumor-expressed antigens have demonstrated clinical efficacy in the treatment of cancer. To broaden the applicability of this... Show moreInfusions of T cells engineered with TCRs directed against tumor-expressed antigens have demonstrated clinical efficacy in the treatment of cancer. To broaden the applicability of this approach a broad repertoire of clinically relevant TCRs is required. This thesis describes a high throughput methodology for the isolation of TCRs useful in the treatment of patients suffering from B-cell malignancies. To induce strong immune responses against tumor-expressed antigens, the immunogencity of allogeneic (non-self) HLA molecules is exploited. Around this fundamental knowledge about T-cell reactivity a pipeline is developed that consists of the selection of targetable tumor-expressed antigens by mining gene expression databases, discovery of HLA-presented peptides from peptide elution studies describing the HLA ligandome of B lymphocytes, a peptide-MHC tetramer based high throughput methodology for the isolation and characterization of T cells expressing TCRs specific for tumor-expressed antigens. Several TCRs were isolated targeting antigens that are expressed on several B-cell malignancies including multiple myeloma. This is of particular interest since novel immunotherapies for multiple myeloma are currently lacking. The work described in this thesis can significantly broaden the applicability of immunotherapy by providing novel tools and reagents for strategies relying on tumor recognition through antigen presentation in the context of HLA. Show less
