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基于金纳米颗粒的截短型ORF2疫苗对小鼠抗戊型肝炎病毒的免疫原性。

Immunogenicity of gold nanoparticle-based truncated ORF2 vaccine in mice against Hepatitis E virus.

作者信息

Rani Dibya, Nayak Baibaswata, Srivastava Sudha

机构信息

Department of Biotechnology, Jaypee Institute of Information Technology, Noida, Uttar Pradesh 201309 India.

Department of Gastroenterology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, 110029 India.

出版信息

3 Biotech. 2021 Feb;11(2):49. doi: 10.1007/s13205-020-02573-y. Epub 2021 Jan 11.

Abstract

This study presents nanoparticle-based vaccine development for Hepatitis E virus (HEV). Gold nanoparticles (GNP) of average size 12 nm were synthesized by citrate reduction method followed by functionalization with cysteamine hydrochloride for nano-conjugation. Immune response of nano-conjugates of GNP with 26 kDa protein (368-606 amino acids) and 54 kDa protein (112-606 amino acids) were evaluated. In vitro release kinetics of GNP-conjugated 54 kDa (GNP54) and 26 kDa (GNP26) proteins showed slower rate of release of 54 kDa protein as compared to 26 kDa protein. Humoral immune response of mice immunized intramuscularly with GNP54, GNP26 and GNP alone, exhibited HEV-specific IgG titer of 7.9 ± 2.9, 5.686 ± 4.098 and 0.698 ± 0.089, respectively, after 14 days of booster immunization. In addition to this, HEV-specific cell-mediated immune response was demonstrated by splenocyte proliferation assay. Analysis of results using one-way ANOVA, showed statistically significant ( value < 0.05) increase in splenocyte proliferation for GNP54- and GNP26-immunized mice in comparison to GNP alone immunized mice. Stimulation index of HEV ORF2 proteins in GNP54/GNP26-immunized mice were comparable to Concanavalin A-treated positive control. These results indicate GNP-based vaccine as a promising candidate for efficiently mediating both humoral and cell-mediated immune response against HEV.

摘要

本研究介绍了基于纳米颗粒的戊型肝炎病毒(HEV)疫苗的开发。通过柠檬酸盐还原法合成平均尺寸为12纳米的金纳米颗粒(GNP),随后用盐酸半胱胺进行功能化以实现纳米共轭。评估了GNP与26 kDa蛋白(368 - 606个氨基酸)和54 kDa蛋白(112 - 606个氨基酸)的纳米共轭物的免疫反应。GNP共轭的54 kDa(GNP54)和26 kDa(GNP26)蛋白的体外释放动力学表明,与26 kDa蛋白相比,54 kDa蛋白的释放速率较慢。用GNP54、GNP26和单独的GNP肌肉注射免疫的小鼠,在加强免疫14天后,其体液免疫反应分别表现出HEV特异性IgG滴度为7.9±2.9、5.686±4.098和0.698±0.089。除此之外,通过脾细胞增殖试验证明了HEV特异性细胞介导的免疫反应。使用单因素方差分析对结果进行分析,结果显示,与单独用GNP免疫的小鼠相比,用GNP54和GNP26免疫的小鼠脾细胞增殖有统计学显著增加(p值<0.05)。GNP54/GNP26免疫的小鼠中HEV ORF2蛋白的刺激指数与伴刀豆球蛋白A处理的阳性对照相当。这些结果表明基于GNP的疫苗是有效介导针对HEV的体液免疫和细胞介导免疫反应的有前景的候选疫苗。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3c5/7801584/2db6cb709b55/13205_2020_2573_Fig1_HTML.jpg

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