The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies
| Autor: | Chizuru Yamamoto, Masataka Nakamura, M Iwasaki, Hiroaki Mizukami, Hiroyuki Ido, Masashi Urabe, Tomonori Tsukahara, Takeshi Teruya, Koichi Kawakami, Yasushi Saga, Akihiro Kume, Renier J. Brentjens, Keiya Ozawa, Ryosuke Uchibori, N Iwase, Ken Ohmine |
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| Rok vydání: | 2014 |
| Předmět: |
Adoptive cell transfer
Lymphoma B-Cell Recombinant Fusion Proteins T-Lymphocytes Transgene Antigens CD19 Molecular Sequence Data Receptors Antigen T-Cell chemical and pharmacologic phenomena Immunotherapy Adoptive Article CD19 Viral vector Mice Cell Line Tumor hemic and lymphatic diseases Genetics Animals Humans Molecular Biology Transposase Mice Knockout Mice Inbred BALB C biology hemic and immune systems Genetic Therapy Molecular biology Coculture Techniques Chimeric antigen receptor Raji cell DNA Transposable Elements NIH 3T3 Cells biology.protein Molecular Medicine Transposon mutagenesis Genetic Engineering Neoplasm Transplantation |
| Zdroj: | Gene Therapy. 22:209-215 |
| ISSN: | 1476-5462 0969-7128 |
| DOI: | 10.1038/gt.2014.104 |
| Popis: | Engineered T-cell therapy using a CD19-specific chimeric antigen receptor (CD19-CAR) is a promising strategy for the treatment of advanced B-cell malignancies. Gene transfer of CARs to T-cells has widely relied on retroviral vectors, but transposon-based gene transfer has recently emerged as a suitable nonviral method to mediate stable transgene expression. The advantages of transposon vectors compared with viral vectors include their simplicity and cost-effectiveness. We used the Tol2 transposon system to stably transfer CD19-CAR into human T-cells. Normal human peripheral blood lymphocytes were co-nucleofected with the Tol2 transposon donor plasmid carrying CD19-CAR and the transposase expression plasmid and were selectively propagated on NIH3T3 cells expressing human CD19. Expanded CD3(+) T-cells with stable and high-level transgene expression (~95%) produced interferon-γ upon stimulation with CD19 and specifically lysed Raji cells, a CD19(+) human B-cell lymphoma cell line. Adoptive transfer of these T-cells suppressed tumor progression in Raji tumor-bearing Rag2(-/-)γc(-/-) immunodeficient mice compared with control mice. These results demonstrate that the Tol2 transposon system could be used to express CD19-CAR in genetically engineered T-cells for the treatment of refractory B-cell malignancies. |
| Databáze: | OpenAIRE |
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