RT Journal Article
SR Electronic
T1 HA Nanocarriers Mediate HepG2 Vaccine Induction of hGM-CSF Gene Antitumor effect study
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 636316
DO 10.1101/636316
A1 Leah Robert
A1 David Gupta
YR 2019
UL http://biorxiv.org/content/early/2019/05/13/636316.abstract
AB This report developed a novel method to observe the anti-tumor effect of HA nanoparticle carrier-mediated HepG2 cell vaccine transfected with hGM-CSF gene in vitro, and provide evidence for the clinical application of hGM-CSF gene-modified HepG2 cell vaccine. HA nanoparticle-mediated hGM-CSF gene transfection of HepG2 cells was used to prepare HepG2 cell vaccine transfected with GMCSF gene. Human PBMC were isolated by density gradient centrifugation and human PBMC were induced in vitro. The proliferative activity of PBMC and the killing effect on HepG2 cells were determined by WST-1 method. The positive expression rates of CD4+ and CD8+ were analyzed by flow cytometry, and the secretion of INF-γ was determined by ELISA. WST-1 results showed that the transgenic HepG2 vaccine induced PBMC proliferation, and its proliferation rate was better than that of wild-type vaccine. The induced PBMC had a higher killing rate against HepG2 than the wild-type vaccine group and each blank. In the control group, FCM results showed that the positive expression rates of CD4+ and CD8+ in the transgenic HepG2 vaccine group were higher than those in the wild-type vaccine group and each blank control group. The ELISA results showed that the IFN-γ content in the transgenic PBMC culture supernatant was 1989.76. +/− 254.21 pg/ml, higher than the wild-type vaccine group and each blank control group.