T-cell receptor transfer into human T cells with ecotropic retroviral vectors.
Adoptive T-cell transfer for cancer immunotherapy requires genetic modification of T cells with recombinant T-cell receptors (TCRs). Amphotropic retroviral vectors (RVs) used for TCR transduction for this purpose are considered safe in principle. Despite this, TCR-coding and packaging vectors could theoretically recombine to produce replication competent vectors (RCVs), and transduced T-cell preparations must be proven free of RCV. To eliminate the need for RCV testing, we transduced human T cells with ecotropic RVs so potential RCV would be non-infectious for human cells. We show that transfection of synthetic messenger RNA encoding murine cationic amino-acid transporter 1 (mCAT-1), the receptor for murine retroviruses, enables efficient transient ecotropic transduction of human T cells. mCAT-1-dependent transduction was more efficient than amphotropic transduction performed in parallel, and preferentially targeted naive T cells. Moreover, we demonstrate that ecotropic TCR transduction results in antigen-specific restimulation of primary human T cells. Thus, ecotropic RVs represent a versatile, safe and potent tool to prepare T cells for the adoptive transfer.
III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
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Koste
TRON (Translational Oncology at the University Medical Center), Johannes Gutenberg University Mainz, Mainz, Germany.
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Beissert
TRON (Translational Oncology at the University Medical Center), Johannes Gutenberg University Mainz, Mainz, Germany.
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H
Hoff
III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
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Pretsch
III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
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Türeci
1] III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany [2] TRON (Translational Oncology at the University Medical Center), Johannes Gutenberg University Mainz, Mainz, Germany.
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Sahin
D019264:Adoptive Transfer; D000818:Animals; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000637:transplantation; D018414:CD8-Positive T-Lymphocytes / Q000166:cytology* / Q000637:transplantation; D027201:Cationic Amino Acid Transporter 1 / Q000235:genetics*; D002460:Cell Line; D018274:Electroporation; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D016219:Immunotherapy, Adoptive; D019169:Jurkat Cells; D017989:Leukemia Virus, Gibbon Ape / Q000235:genetics; D008562:Membrane Glycoproteins / Q000235:genetics; D051379:Mice; D010957:Plasmids / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics*; D014161:Transduction, Genetic; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000235:genetics
D027201:Cationic Amino Acid Transporter 1; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D014759:Viral Envelope Proteins