Bacterial outer membrane vesicles as nano carriers to study immunological activities.
- Author:
Chen QI
1
;
W U MIN
1
;
Bai HONGZHEN
1
;
Guo ZELING
1
;
Zhou JUN
1
;
Wang QINGQING
1
;
Tang GUPING
2
Author Information
1. Department of Chemistry, Zhejiang University, Hangzhou 310028, China.
2. Department of Chemistry, Zhejiang University, Hangzhou 310028, China. tangguping@zju.edu.cn.
- Publication Type:Journal Article
- From:
Journal of Zhejiang University. Medical sciences
2017;46(2):144-150
- CountryChina
- Language:Chinese
-
Abstract:
To prepare a nano-carrier based on combining bacterial outer membrane vesicles (OMV) with three block polymer pluronic F127 (PEO-PPO-PEO) (OMV-F127) and to investigate its immunological activity.Attenuated salmonella (sal) was cultivated. OMV were separated by centrifugal ultrafiltration or ultrasonication, and OMV-F127 was prepared by mechanical extrudation method. The protein contents and compositions were tested with BCA and SDS-PAGE; the morphology of OMV, F127 and OMV-F127 were observed with FM and TEM; the particle sizes and their zeta potential were determined with DLS. Mouse macrophage RAW246.7 cells were treated with OMV-F127 (50 μg/mL, 100 μg/mL) in vitro, and the concentrations of IL-12, TNF-α and IFN-γ in culture supernatant were measured with ELISA kits.The contents of protein in separated OMV by centrifugal ultrafiltration and ultrasonication were 2.8 mg/mL and 2.7 mg/mL, respectively. SDS-PAGE showed the marker protein OmpF/C in OMV. Under the FM and TEM, ball-like structure of F127 and OMV-F127 was observed. Size analysis revealed that the diameters of OMV, F127 and OMV-F127 were 72±2 nm, 90±3 nm and 92±2 nm, respectively. ELISA tests revealed that OMV-F127 significantly stimulated the secretion of IL-12, TNF-α and IFN-γ in RAW246.7 cells.A nano-carrier based on bacterial outer membrane vesicles has been prepared, which can stimulate the secretion of cytokines and may have immunomodulatory effects.
