Details
- Autor(en) / Beteiligte
- Titel
- A transgene-encoded cell surface polypeptide for selection, in vivo tracking, and ablation of engineered cells
- Ist Teil von
- Blood, 2011-08, Vol.118 (5), p.1255-1263
- Ort / Verlag
- Washington, DC: Elsevier Inc
- Erscheinungsjahr
- 2011
- Link zum Volltext
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- An unmet need in cell engineering is the availability of a single transgene encoded, functionally inert, human polypeptide that can serve multiple purposes, including ex vivo cell selection, in vivo cell tracking, and as a target for in vivo cell ablation. Here we describe a truncated human EGFR polypeptide (huEGFRt) that is devoid of extracellular N-terminal ligand binding domains and intracellular receptor tyrosine kinase activity but retains the native amino acid sequence, type I transmembrane cell surface localization, and a conformationally intact binding epitope for pharmaceutical-grade anti-EGFR monoclonal antibody, cetuximab (Erbitux). After lentiviral transduction of human T cells with vectors that coordinately express tumor-specific chimeric antigen receptors and huEGFRt, we show that huEGFRt serves as a highly efficient selection epitope for chimeric antigen receptor+ T cells using biotinylated cetuximab in conjunction with current good manufacturing practices (cGMP)-grade anti-biotin immunomagnetic microbeads. Moreover, huEGFRt provides a cell surface marker for in vivo tracking of adoptively transferred T cells using both flow cytometry and immunohistochemistry, and a target for cetuximab-mediated antibody-dependent cellular cytotoxicity and in vivo elimination. The versatility of huEGFRt and the availability of pharmaceutical-grade reagents for its clinical application denote huEGFRt as a significant new tool for cellular engineering.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0006-4971eISSN: 1528-0020DOI: 10.1182/blood-2011-02-337360Titel-ID: cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3152493
- Format
- –
- Schlagworte
- Animals, Antigens, Surface - genetics, Apoptosis - genetics, Biological and medical sciences, Cell Separation - methods, Cell Tracking - methods, Cells - metabolism, Cells - pathology, Cells, Cultured, Gene Therapy, Genes, erbB-1, Genes, Reporter, Genes, Transgenic, Suicide - physiology, Hematologic and hematopoietic diseases, Humans, Interleukin Receptor Common gamma Subunit - genetics, Medical sciences, Mice, Mice, Inbred NOD, Mice, SCID, Mice, Transgenic, Peptides - genetics, Tissue Engineering - methods, Transgenes - genetics