Acta Biomater. 2020 Apr 25. pii: S1742-7061(20)30218-X. doi: 10.1016/j.actbio.2020.04.021. [Epub ahead of print]
Genetic immunization against hepatitis B virus with calcium phosphate nanoparticles in vitro and in vivo.
Rojas-Sánchez L1, Zhang E2, Sokolova V1, Zhong M2, Yan H2, Lu M3, Li Q3, Yan H4, Epple M5.
Author information
1
Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitaetsstr. 5-7, 45117 Essen, Germany.
2
Mucosal Immunity Research Group, State Key Laboratory of Virology, Wuhan Institute of Virology (WIV), Chinese Academy of Sciences (CAS), Wuhan 430071, P.R. China.
3
Institute of Virology, University Hospital Essen, University of Duisburg-Essen, Hufelandstr. 55, 45147 Essen, Germany.
4
Mucosal Immunity Research Group, State Key Laboratory of Virology, Wuhan Institute of Virology (WIV), Chinese Academy of Sciences (CAS), Wuhan 430071, P.R. China. Electronic address: [email protected].
5
Inorganic Chemistry and Center for Nanointegration Duisburg-Essen (CeNIDE), University of Duisburg-Essen, Universitaetsstr. 5-7, 45117 Essen, Germany. Electronic address: [email protected].
Abstract
Calcium phosphate nanoparticles were loaded with plasmid DNA and toll-like receptor ligands (TLR), i.e. CpG or flagellin, to activate antigen-presenting cells (APCs) like dendritic cells (DCs). The functionalized nanoparticles were studied in vitro on HeLa, C2C12 and BHK-21 cell lines, focusing on the expression of two specific proteins. EGFP-DNA, encoding for enhanced green fluorescent protein (EGFP), was used as a model plasmid to optimize the transfection efficiency in vitro by fluorescence microscopy and flow cytometry. Calcium phosphate nanoparticles loaded with TLR ligands and plasmid DNA encoding for the hepatitis B virus surface antigen (pHBsAg) were evaluated by in vitro and in vivo immunization experiments to identify a possible candidate for a prophylactic hepatitis B virus (HBV) vaccine. The nanoparticles induced a strong expression of HBsAg in the three cell lines. In splenocytes, the expression of the co-stimulatory molecules CD80 and CD86 was enhanced. After intramuscular injection in mice, the nanoparticles induced the expression of HBsAg, the antigen-specific T cell response, and the antigen-specific antibody response (IgG1).