Transgenic IL-2 expression in Ewing tumor cell lines after transfection with Starburst dendrimers and cationic liposomes.
Immunotherapy with IL-2-transduced cells requires efficient methods of gene transfer. Nonviral methods have been studied intensively in recent years. This study examined whether tumor and fibroblast cell lines established from Ewing tumor patients could be efficiently transfected with the IL-2 gene. Starburst dendrimers (Superfect), a novel transfection reagent, were chosen for a transfection study and optimal conditions for gene transfer were evaluated. Three Ewing tumor cell lines and 3 fibroblast cell lines obtained from Ewing tumor patients were analyzed. The concentration of IL-2 in the supernatant of transfected cells was measured by ELISA. All three Ewing tumor cell lines transfected by Starburst dendrimers yielded higher IL-2 levels than after lipofection. In contrast to lipofection, expression of IL-2 increased with time and peaked later. In one of three tested fibroblast cell lines, transfection using Superfect yielded higher IL-2 levels. IL-2 production was generally lower in fibroblasts as compared to Ewing tumor cell lines. Given the low toxicity of Superfect reagent and the high efficiency of transfection, this method seems to be ideal for clinical studies on the immunotherapy of Ewing tumors.
Ackermann
Birgit
B
Laboratory for Experimental Hematology and Stem Cell Transplantation, Department of Pediatric Hematology/Oncology, Heinrich-Heine University of Düsseldorf, Düsseldorf, Germany.
Engel
Barbara C
BC
Buttlies
Barbara
B
Zibert
Andree
A
Burdach
Stefan
S
D002412:Cations; D050091:Dendrimers; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007167:Immunotherapy / Q000379:methods; D007376:Interleukin-2 / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D008081:Liposomes / Q000493:pharmacokinetics; D011073:Polyamines / Q000493:pharmacokinetics; D012512:Sarcoma, Ewing / Q000473:pathology*; D014162:Transfection / Q000379:methods* / Q000592:standards; D014407:Tumor Cells, Cultured
D002412:Cations; D050091:Dendrimers; D007376:Interleukin-2; D008081:Liposomes; C104700:PAMAM Starburst; D011073:Polyamines