在体外和体内用磷酸钙纳米粒子对乙型肝炎病毒进行基因免

StephenW

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).

Copyright © 2020. Published by Elsevier Ltd.
KEYWORDS:

Calcium phosphate; Hepatitis B virus; Immunization; Nanomedicine; Nanoparticles; Virology

PMID:
    32344172
DOI:
    10.1016/j.actbio.2020.04.021

StephenW

Acta Biomater。 2020年4月25日。pii：S1742-7061（20）30218-X。 doi：10.1016 / j.actbio.2020.04.021。 [Epub提前发行]
在体外和体内用磷酸钙纳米粒子对乙型肝炎病毒进行基因免疫。
Rojas-SánchezL1，Zhang E2，Sokolova V1，Zhong M2，Yan H2，Lu M3，Li Q3，Yan H4，Epple M5。
作者信息

1个
    杜伊斯堡-埃森大学，无机化学与纳米整合中心杜伊斯堡-埃森（CeNIDE），Universitaetsstr。 5-7，45117埃森，德国。
2
    中国科学院武汉病毒研究所（WIV）病毒学国家重点实验室粘膜免疫研究小组，中国武汉430071。
3
    杜伊斯堡埃森大学埃森大学医院病毒研究所，Hufelandstr。 55，45147 Essen，Germany。
4
    中国科学院武汉病毒研究所（WIV）病毒学国家重点实验室粘膜免疫研究小组，中国武汉430071。电子地址：[email protected]。
5
    杜伊斯堡-埃森大学，无机化学与纳米整合中心杜伊斯堡-埃森（CeNIDE），Universitaetsstr。 5-7，45117埃森，德国。电子地址：[email protected]。

抽象

磷酸钙纳米颗粒上装有质粒DNA和Toll样受体配体（TLR），即CpG或鞭毛蛋白，以激活树突状细胞（DC）等抗原呈递细胞（APC）。在HeLa，C2C12和BHK-21细胞系上体外研究了功能化的纳米颗粒，重点研究了两种特定蛋白质的表达。编码增强型绿色荧光蛋白（EGFP）的EGFP-DNA被用作模型质粒，以通过荧光显微镜和流式细胞术优化体外转染效率。通过体外和体内免疫实验评估了装载有TLR配体的磷酸钙纳米颗粒和编码乙型肝炎病毒表面抗原（pHBsAg）的质粒DNA，以确定预防性乙型肝炎疫苗的可能候选者。纳米粒子在三种细胞系中诱导了HBsAg的强烈表达。在脾细胞中，共刺激分子CD80和CD86的表达增强。小鼠肌肉内注射后，纳米颗粒诱导HBsAg的表达，抗原特异性T细胞反应和抗原特异性抗体反应（IgG1）。

版权所有©2020。由Elsevier Ltd.发布。
关键字：

磷酸钙；乙型肝炎病毒;免疫；纳米医学纳米颗粒；病毒学

PMID：
    32344172
DOI：
    10.1016 / j.actbio.2020.04.021