A novel 'sort-suicide' fusion gene vector for T cell manipulation.
Retroviral suicide gene vectors have successfully been used in clinical studies to improve the safety of adoptive immunotherapy with allogeneic T lymphocytes in the treatment of malignant and viral diseases. At the same time these studies have revealed several problems that are yet to be resolved including impaired T cell function due to long ex vivo culture. Here we present new retroviral vectors co-expressing truncated CD34, a gene transfer marker which ensures rapid enrichment of transduced cells using commercially available GMP-approved devices, and a splice-corrected variant of Herpes simplex virus thymidine kinase (scHSVtk) which confers high sensitivity to the prodrug ganciclovir. We show that a retroviral hybrid vector, MP71, based on the myeloproliferative sarcoma virus (MPSV) and the murine embryonic stem cell virus (MESV), encoding a tCD34/scHSVtk fusion protein mediates high expression of the 'sort-suicide' selection marker, thereby allowing for highly efficient purification and selective elimination of transduced cells.
Fehse
B
B
Bone Marrow Transplantation, University Hospital Hamburg-Eppendorf, Germany.
Kustikova
O S
OS
Li
Z
Z
Wahlers
A
A
Bohn
W
W
Beyer
W R
WR
Chalmers
D
D
Tiberghien
P
P
Kühlcke
K
K
Zander
A R
AR
Baum
C
C
D018952:Antigens, CD34 / Q000235:genetics; D015774:Ganciclovir / Q000494:pharmacology; D005819:Genetic Markers; D005822:Genetic Vectors; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D019169:Jurkat Cells; D008212:Lymphocyte Depletion / Q000379:methods*; D018613:Microscopy, Confocal; D012190:Retroviridae / Q000235:genetics; D013601:T-Lymphocytes / Q000187:drug effects / Q000637:transplantation*; D014161:Transduction, Genetic
D018952:Antigens, CD34; D005819:Genetic Markers; D015774:Ganciclovir