Bacterial outer membrane vesicles as nano carriers to study immunological activities
10.3785/j.issn.1008-9292.2017.04.02
- VernacularTitle:细菌外膜囊泡纳米载体的制备及其免疫调节作用
- Author:
Qi CHEN
1
;
Min WU
;
Hongzhen BAI
;
Zeling GUO
;
Jun ZHOU
;
Qingqing WANG
;
Guping TANG
Author Information
1. 浙江大学化学系
- Keywords:
Salmonella vaccines;
Adjuvants,immunologic;
Bacterial outer membrane proteins;
Nanostructures;
Polymers/pharmacology;
Macrophages;
Cytokines;
Cells,cultured
- From:
Journal of Zhejiang University. Medical sciences
2017;46(2):118-124
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare a nano-carrier based on combining bacterial outer membrane vesicles (OMV) with three block polymer pluronic F127 (PEO100-PPOss-PEO100) (OMV-F127) and to investigate its immunological activity.Methods: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-γin culture supernatant were measured with ELISA kits.Results: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.Conclusion:A nano-carrier based on bacterial OMV has been prepared,which can stimulate the secretion of cytokines and may have immunomodulatory effects.
