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• W1637441631 abstract "AACR Annual Meeting-- Apr 18-22, 2009; Denver, COPurpose: It is well documented that activation, expansion and differentiation of naive T lymphocytes results in the appearance of perforin+/granzyme B+/IFN-gamma+ effectors. This study examined whether expanded T lymphocytes may radiosensitize tumor cells through the release of IFN-gamma, and whether CD4+ and CD8+ T cells display distinct IFN-gamma and radiosensitizing profiles. Methods: Naive T lymphocytes from Balb/c mice were activated with IL12 plus IL18. T-cell expansion was achieved by a 2-day priming with bound anti-CD3 plus soluble anti-CD28 antibodies, further growth in the presence of IL-2 for 7 days, and restimulation through CD3/CD28. CD8+ and CD4+ T cells were selected by immunomagnetic MACS beads and analyzed for intracellular IFN-gamma by FACS, RT-PCR and ELISA. To radiosensitize target EMT-6 tumor cells, subconfluent cultures were incubated with activated T cells or their conditioned medium (CM), irradiated at 5-15 Gy in 1% oxygen, and reseeded for colony formation assay. The expression of inducible nitric oxide synthase (iNOS), a downstream target of IFN-gamma in tumor cells, was analyzed by RT-PCR and production of NO/nitrite. Results: Expanded and restimulated T cells displayed a perforin+/granzyme B+/IFN-gamma+ phenotype, and enhanced the radiosensitivity of hypoxic EMT-6 cells up to 1.8-fold at an effector/target ratio 1:1. Similar effects were induced by purified CD8+ T cells, which produced IFN-gamma at high levels (2.9 ng/103 cells), and expressed 80-90% IFN-gamma+ phenotype. In contrast, CD4+ T cells revealed reduced levels (by 3-5-times) of IFN-gamma expression and secretion, and a 1.4-fold radiosensitization only. Similar radiosensitizing effects could be induced by CM diluted 1:30, and were abrogated by anti-IFN-gamma antibodies and by aminoguanidine, a metabolic inhibitor of iNOS. These results indicate that the observed radiosensitization does not require direct immune-tumor cell interaction, but is linked to the release of IFN-gamma, a potent activator of iNOS in tumor cells. In line, iNOS up-regulation in EMT-6 cells was confirmed at transcriptional and protein levels. Naive T cells secreted marginal levels of IFN-gamma+ and did not affect the radiosensitivity of EMT-6 tumor cells. Following activation with IL12/IL18, naive T cells showed 8-10% IFN-gamma+ phenotype, and were able to radiosensitize EMT-6 cells at an effector/target ratio 10:1, or at a dilution of CM 1:3. Conclusion: The radiosensitizing potential of activated T cells is primarily linked to IFN-gamma+ CD8+ T cells, and is drastically increased along T-cell expansion.Citation Information: In: Proc Am Assoc Cancer Res; 2009 Apr 18-22; Denver, CO. Philadelphia (PA): AACR; 2009. Abstract nr 2306." @default.
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• W1637441631 date "2009-05-01" @default.
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• W1637441631 title "Abstract #2306: Radiosensitizing potential of T lymhocytes: expanded CD4+ and CD8+ T cells display distinct patterns of IFN-gamma expression and radiosensitizing activity" @default.
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