| Abstract: |
The functional activity and feasibility of noninvasive monitoring of tumor-specific T-cells, transduced with a reporter gene of human origin, were evaluated. Donor T-cells were transduced with a chimeric antigen receptor (CAR) specific for human prostate-specific membrane antigen (hPSMA) and a new mutant of human deoxycitidine kinase (dCKDM) reporter gene. The sensitivities of transduced T-cells to gancyclovir and cytarabine were evaluated by the WST-test (measurements of mitochondrial dehydrogenase activity), specific cytotoxicity of transduced T-cells was evaluated by aCella-TOX method (measurement of glyceraldehyde-3-phosphate dehydrogenase release in vitro). Lung prostate cancer tumors formed in mice after injection of human prostatic cancer cells, expressing hPSMA. Positron emission tomography (PET) with 2-fluoro-2’deoxy-1-beta-D-arabinofuranoside-5-ethyluracyl, labeled by18 F (18 F-FEAU) isotope, was carried out on the day of T-cell injection and on day 3 after injection. The expression of anti-hPSMA CAR and dCKDM genes was detected in 75% transduced lymphocytes. Co-transduction of T-cells with CAR and dCKDM reporter gene did not affect their cytolytic activity against hPSMA+-target cells. T-cells transduced with dCKDM reporter gene were sensitive to cytarabine but not gancyclovir. PET imaging revealed distinct accumulation of T-cells in lung tumors of animals after intravenous injection of dCKDM-transduced lymphocytes on the day of injection and on day 3 after it, which was confirmed by the immunohistochemical method. © 2012, Fund - Doctors, Innovations,Science for Children. All rights reserved. |
