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Journal of Acquired Immune Deficiency Syndromes & Human Retrovirology:
National AIDS Malignancy Conference

EXPANSION OF EPSTEIN-BARR VIRUS (EBV)-SPECIFIC CYTOTOXIC T CELLS (CTLs) IN GAS PERMEABLE CELL CULTURE BAGS: 127

Davis, J. M.; Lemas, M. V.; Orentas, R.; Eby, L. L.; Fabian, D.; Flinn, I.; Noga, S. J.; Ambinder, R. F.

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The Johns Hopkins Oncology Center. Baltimore, Maryland

EBV-specific CTLs (CD8 effector T cells) cultured in 24-well plates have been used to treat post-transplant lymphoproliferative disease or prevent its development in childhood recipients of bone marrow transplants. Expansion in"open" systems, particularly in multiwell plates, is cumbersome and associated with risks of contamination. Therefore we have investigated the feasibility of expansion of EBV CTL's in closed gas-permeable bags.

Methods: Lymphoblastoid cell lines (LCL) were produced by immortalizing peripheral blood lymphocytes with EBV. Autologous mononuclear cells collected by apheresis were incubated with irradiated LCL's. These incubations were set up in parallel in 24 well plates and in gas-permeable polyolefin-polystyrene bags (PL2417) from Baxter. After 3 weeks of stimulation with weekly addition of irradiated LCL's. IL-2 (25 units/ml) was added to the media. On day 28 the expanded products were evaluated. The T cell products expanded in bags were administered to patients in a pilot study.

Results: Expansion of cells in 24 well plates required 19 and 27 plates versus 2 bags. EBV-specific cytolytic activity was evaluated by chromium release assay using autologous LCL's as targets. At various effector to target ratios lytic activity against autologous LCLs were similar. Flow cytometric analysis also showed similar profiles with >95% of cells marking with CD3 and >80% of cells marking with CD8. Two patients were treated with weekly CTL infusions(107/m2) for four consecutive weeks without adverse effects.

Conclusions: Gas-permeable polyolefin polystyrene bags are suitable for use in the expansion of EBV-specific CTLs for clinical purposes.

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National AIDS Malignancy Conference

Bethesda, Maryland April 28-30, 1997

sponsored by the National Cancer Institute

© Lippincott-Raven Publishers.

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