SVCV纳米疫苗的传递和跨膜转运机制

Han Zhang , Pan-pan Zhang , Xin-Xin Liu , Hai-hua Peng , Qun Liu , Jun Wang , Bin Zhu , Yong-Can Zhou , Yun Sun , Chen Zhang
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摘要

鲤鱼病毒春季病毒血症(SVCV)是多种鲤科鱼类的致命微生物,在淡水鱼养殖中造成巨大的经济损失。为了减轻损失,浸入式疫苗接种是预防和控制SVCV感染的极有希望的方法。在我们之前的研究中,我们设计了一种针对SVCV的模块化浸泡纳米疫苗(LSG-TDH),发现它不仅可以诱导斑马鱼的粘膜免疫,还可以诱导全身免疫反应。然而,我们之前的研究无法详细说明LSG-TDH是如何被细胞吸收的。因此,本研究以cyprini丘疹上皮瘤(EPC)细胞和巨噬细胞为模型,探讨其体外跨膜转运机制。通过细胞荧光、流式细胞术和化学抑制剂实验分析LSG-TDH对EPC细胞和巨噬细胞的跨膜转运机制。结果表明,LSG-TDH进入细胞具有时间依赖性和能量依赖性。氯丙嗪显著抑制LSG-TDH的跨膜转运。氯丙嗪处理后,巨噬细胞和巨噬细胞中LSG-TDH含量分别下降80 %和71 %。提示LSG-TDH的跨膜转运主要通过网格蛋白介导的胞吞途径实现。本研究为水生纳米疫苗的递送机制提供了参考,具有重要的科学意义和应用前景。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
The delivery and transmembrane transport mechanism of SVCV nanovaccine
Spring viremia of carp virus (SVCV) is a fatal microorganism for a variety of cyprinid fish species, sparking off enormous economic losses in freshwater fish aquaculture. For alleviating losses, immersion vaccination is an extremely promising maneuver to prevent and curb on SVCV infection. In our previous research, we designed a modular immersion nanovaccine (LSG-TDH) targeting SVCV, and found that it could induce not merely mucosal immunity but systemic immune responses in zebrafish. Nevertheless, our previous research was unable to elaborate on how LSG-TDH were taken up by the cells. Thereby, this study aimed to investigate its transmembrane transport mechanism in vitro using epithelioma papulosum cyprini (EPC) cells and macrophages as models. The transmembrane transport mechanism of LSG-TDH on EPC cells and macrophages was analyzed by cell fluorescence, flow cytometry and chemical inhibitor experiments. The results showed that the entry of LSG-TDH into cells was time-dependent and energy-dependent. Chlorpromazine significantly inhibited the transmembrane transport of LSG-TDH. The contents of LSG-TDH into EPC cells and macrophages after treated with chlorpromazine decreased by 80 % and 71 %, respectively. It is indicated that the transmembrane transport of LSG-TDH was mainly achieved through clathrin-mediated endocytosis pathway. This study provides a reference for the delivery mechanism of aquatic nanovaccine, which has important scientific significance and application prospect.
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