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明胶纳米颗粒增强丙型肝炎病毒重组NS2基因的递送。

Gelatin nanoparticles enhance delivery of hepatitis C virus recombinant NS2 gene.

作者信息

Sabet Salwa, George Marina A, El-Shorbagy Haidan M, Bassiony Heba, Farroh Khaled Y, Youssef Tareq, Salaheldin Taher A

机构信息

Department of Zoology, Faculty of Science, Cairo University, Giza, Egypt.

Department of Photochemistry and Photobiology, National Institute of Laser Enhanced Sciences, Cairo University, Giza, Egypt.

出版信息

PLoS One. 2017 Jul 26;12(7):e0181723. doi: 10.1371/journal.pone.0181723. eCollection 2017.

Abstract

BACKGROUND

Development of an effective non-viral vaccine against hepatitis C virus infection is of a great importance. Gelatin nanoparticles (Gel.NPs) have an attention and promising approach as a viable carrier for delivery of vaccine, gene, drug and other biomolecules in the body.

AIM OF WORK

The present study aimed to develop stable Gel.NPs conjugated with nonstructural protein 2 (NS2) gene of Hepatitis C Virus genotype 4a (HCV4a) as a safe and an efficient vaccine delivery system.

METHODS AND RESULTS

Gel.NPs were synthesized and characterized (size: 150±2 nm and zeta potential +17.6 mv). NS2 gene was successfully cloned and expressed into E. coli M15 using pQE-30 vector. Antigenicity of the recombinant NS2 protein was confirmed by Western blotting to verify the efficiency of NS2 as a possible vaccine. Then NS2 gene was conjugated to gelatin nanoparticles and a successful conjugation was confirmed by labeling and imaging using Confocal Laser Scanning Microscope (CLSM). Interestingly, the transformation of the conjugated NS2/Gel.NPs complex into E. coli DH5-α was 50% more efficient than transformation with the gene alone. In addition, conjugated NS2/Gel.NPs with ratio 1:100 (w/w) showed higher transformation efficiency into E. coli DH5-α than the other ratios (1:50 and 2:50).

CONCLUSION

Gel.NPs effectively enhanced the gene delivery in bacterial cells without affecting the structure of NS2 gene and could be used as a safe, easy, rapid, cost-effective and non-viral vaccine delivery system for HCV.

摘要

背景

开发一种有效的抗丙型肝炎病毒感染的非病毒疫苗具有重要意义。明胶纳米颗粒(Gel.NPs)作为一种可行的载体,用于在体内递送疫苗、基因、药物和其他生物分子,受到关注且前景广阔。

工作目的

本研究旨在开发与丙型肝炎病毒4a基因型(HCV4a)的非结构蛋白2(NS2)基因偶联的稳定明胶纳米颗粒,作为一种安全有效的疫苗递送系统。

方法与结果

合成并表征了明胶纳米颗粒(尺寸:150±2 nm,ζ电位 +17.6 mv)。使用pQE - 30载体成功将NS2基因克隆并表达至大肠杆菌M15中。通过蛋白质免疫印迹法确认重组NS2蛋白的抗原性,以验证NS2作为潜在疫苗的有效性。然后将NS2基因与明胶纳米颗粒偶联,并使用共聚焦激光扫描显微镜(CLSM)进行标记和成像,证实偶联成功。有趣的是,偶联的NS2/Gel.NPs复合物转化至大肠杆菌DH5-α的效率比单独基因转化高50%。此外,比例为1:100(w/w)的偶联NS2/Gel.NPs转化至大肠杆菌DH5-α的效率高于其他比例(1:50和2:50)。

结论

明胶纳米颗粒有效地增强了细菌细胞中的基因递送,且不影响NS2基因的结构,可作为一种安全、简便、快速、经济高效的非病毒丙型肝炎疫苗递送系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a2c2/5528829/4e2f75aaed6b/pone.0181723.g001.jpg

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