title: Soluble T-cell receptors produced in human cells for targeted delivery.
abstract: Recently, technology has become available to generate soluble T-cell receptors (sTCRs) that contain the antigen recognition part. In contrast to antibodies, sTCRs recognize intracellular in addition to extracellular epitopes, potentially increasing the number of applications as reagents for target detection and immunotherapy. Moreover, recent data show that they can be used for identification of their natural peptide ligands in disease. Here we describe a new and simplified expression method for sTCRs in human cells and show that these sTCRs can be used for antigen-specific labeling and elimination of human target cells. Four different TCRs were solubilized by expression of constructs encoding the TCR alpha (α) and beta (β) chains lacking the transmembrane and intracellular domains, linked by a ribosomal skipping 2A sequence that facilitates equimolar production of the chains. Cell supernatants containing sTCRs labeled target cells directly in a peptide (p)-human leukocyte antigen (HLA)-specific manner. We demonstrated that a MART-1p/HLA-A*02:01-specific sTCR fused to a fluorescent protein, or multimerized onto magnetic nanoparticles, could be internalized. Moreover, we showed that this sTCR and two sTCRs recognizing CD20p/HLA-A*02:01 could mediate selective elimination of target cells expressing the relevant pHLA complex when tetramerized to streptavidin-conjugated toxin, demonstrating the potential for specific delivery of cargo. This simple and efficient method can be utilized to generate a wide range of minimally modified sTCRs from the naturally occurring TCR repertoire for antigen-specific detection and targeting.
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway.
authors: Even
authors: E
authors: Walseng
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway; Department of Cell Therapy, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway.
authors: Sébastien
authors: S
authors: Wälchli
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway.
authors: Lars-Egil
authors: LE
authors: Fallang
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway; K. G. Jebsen Center for Cancer Immunotherapy and K.G. Jebsen Center for Inflammation Research, Institute for Clinical Medicine, University of Oslo, Oslo, Norway.
authors: Weiwen
authors: W
authors: Yang
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway; K. G. Jebsen Center for Cancer Immunotherapy and K.G. Jebsen Center for Inflammation Research, Institute for Clinical Medicine, University of Oslo, Oslo, Norway.
authors: Anette
authors: A
authors: Vefferstad
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway.
authors: Ali
authors: A
authors: Areffard
authors: Department of Immunology, Institute for Cancer Research, Oslo University Hospital Radiumhospitalet, Oslo, Norway; K. G. Jebsen Center for Cancer Immunotherapy and K.G. Jebsen Center for Inflammation Research, Institute for Clinical Medicine, University of Oslo, Oslo, Norway.
authors: Johanna
authors: J
authors: Olweus
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular / Q000379:methods*; D016503:Drug Delivery Systems; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D000922:Immunotoxins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D058965:MART-1 Antigen / Q000378:metabolism; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D011948:Receptors, Antigen, T-Cell / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054789:Ribosome Inactivating Proteins, Type 1 / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000076985:Saporins; D012995:Solubility
chemical_list: D000922:Immunotoxins; D058965:MART-1 Antigen; D010455:Peptides; D011948:Receptors, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins