Interaction of fas and fas ligand in a rat 36b10 glioma model

Abstract

Object. An experimental model of the fas and fas ligand (fasL) interaction in malignant glioma was developed. Methods. 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 expres- sion 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, sug- gesting that activation of the transfected fas receptor by fasL expression on the carrier cell was occurring. Conclusion. Based on their experiments, the authors describe a model for evaluating the interaction of fas and fasL in a cellular model of malignant glioma.