Preparation and identification of the renal-cancer vaccine of exosome modified to express GPI-anchored IL-12
Abstract：In the study, we aimed to prepare IL-12-anchored exosomes derived from renal cancer cells and identify their structure and function. Firstly, we constructed a mammalian co-expression plasmid of GPI-anchor-IL-12 by cloning fusion gene of GPI-anchor signal sequence and p40 cDNA in pBudCE4.1. Then we analyzed the expression of fusion protein by using confocal laser scanning microscopy and flow cytometry, and confirmed the IL-12 expression on the cell surface. Ultrafiltration and sucrose gradient centrifugation were employed to purify the exosomes, then TEM and Western blotting showed the exosomes were 30-80 nm in diameter with typical saucer-shape morphology, and expressing HSP70, ICAM-1, G250 and GPI-IL-12. About (80±9.6) pg/ml of IL-12 was detected in 10 n,g/ml exosome which could significantly induce the release of IFN-7. The results suggest that the vaccine of exosomes-GPI-IL-12 can be obtained from the culture supernatant of renal cancer cells modified to express anchored IL-12, which expressing IL-12 and tumor associated antigen G250, and might be a new strategy for treatment of renal cancer.
Keywords：Renal cell carcinomaExosomeInterleukin 12Tumor vaccine
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 344-348 )