An experimental model of the fas and fas ligand (fasL) interaction in malignant glioma was developed.
Using plasmid-based delivery, 36B10 rat glioma cells were modified to express fas (36B10-fas), and a delivery fibroblast cell line was modified to produce fasL, resulting in the FR-fasL cell line. Evaluation of fas expression was performed with flow cytometry and expression of fasL confirmed by Western blot analysis. Once the cell lines were created and partially characterized, fas-induced cytotoxicity was evaluated using an antibody-mediated assay for 36B10-fas that demonstrated significant toxicity at 24 and 48 hours. To evaluate the potential for activating the fas molecule by using cell-mediated delivery, coculture cytotoxicity studies were performed with a target cell line (36B10-fas) and effector cell line (FR-fasL). Using a series of culture ratios, increasing cytotoxicity was noted, suggesting that activation of the transfected fas receptor by fasL expression on the carrier cell was occurring.
Based on their experiments, the authors describe a model for evaluating the interaction of fas and fasL in a cellular model of malignant glioma.