Cytotoxic effects against hepatoma cells by DCs loaded with exosome derived from Huh-7 cells
10.3969/j.issn.1000-484X.2016.04.015
- VernacularTitle:T-exo 负载DC诱导抗肝癌细胞免疫效应研究
- Author:
Jieyu LI
;
Mingshui CHEN
;
Shuping CHEN
;
Zhifeng ZHOU
;
Ling WANG
;
Yunbin YE
- Publication Type:Journal Article
- Keywords:
Dendritic cells(DC);
Exosome;
Liver cancer;
Cytotoxic T cells
- From:
Chinese Journal of Immunology
2016;32(4):519-523
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the cytotoxic effects of CTL cell induced by DCs loaded with exosomes derived from hepatoma Huh-7 cells(T-exo).Methods: Exosomes derived from hepatoma Huh-7 cells were isolated and purified by combination of ultrafiltration centrifugation and sucrose density gradient centrifugation.Morphology of exosomes was observed under transmission electron microscopy and the expression of CD 9,CD63,HSP70 and AFP was detected by Western blot.DCs were induced with peripheral blood monocytes isolated from healthy donors.Flow cytometry was used to analysis surface markers of the DCs loaded with T-exo.WST-1 light absorption measurement was adopted to evaluate the T cell proliferation ability.Annexin-V/PI Flow cytometry were respectively used to examined cytotoxicity against the tumor cells.Results:Exosomes isolated and extracted from culture supernatant of Huh-7 cells presented as circular or elliptical vesicle with bilayer membrane , unequal in size , and with diameter of 50 to 100 nm.Western blot showed that the T-exo expressed CD9,CD63,HSP70 and AFP molecules.DCs loaded with T-exo caused significantly higher T cell pro-liferation and cytotoxic effect against AFP positive Huh-7 cells as compare to gainst AFP negative SMMC 7721 cells and un-loaded control group ( P<0.05 ).Conclusion: T-exosome loaded-DC can promote proliferation and induce significant cytotoxic effect of CTL against Huh-7 cells.
