来源于β-葡聚糖的胶囊作为DNA疫苗的递送系统。

-Derived β-Glucan Capsules as a Delivery System for DNA Vaccines.

作者信息

Pinho Samara Sousa de, Invenção Maria da Conceição Viana, Silva Anna Jéssica Duarte, Macêdo Larissa Silva de, Espinoza Benigno Cristofer Flores, Leal Lígia Rosa Sales, da Gama Marco Antonio Turiah Machado, Moura Ingrid Andrêssa de, Silva Micaela Evellin Dos Santos, Souza Débora Vitória Santos de, Lara Marina Linhares, Alves Julia Nayane Soares Azevedo, Freitas Antonio Carlos de

机构信息

Laboratory of Molecular Studies and Experimental Therapy-LEMTE, Department of Genetics, Federal University of Pernambuco, Recife 50670-901, Brazil.

出版信息

Vaccines (Basel). 2024 Dec 18;12(12):1428. doi: 10.3390/vaccines12121428.

DOI:10.3390/vaccines12121428

PMID:39772088

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11728682/

Abstract

BACKGROUND/OBJECTIVES: DNA vaccines are rapidly produced and adaptable to different pathogens, but they face considerable challenges regarding stability and delivery to the cellular target. Thus, effective delivery methods are essential for the success of these vaccines. Here, we evaluated the efficacy of capsules derived from the cell wall of the yeast as a delivery system for DNA vaccines.

METHODS

The capsules were extracted from the yeast strain GS115, previously grown in a YPD medium. pVAX1 expression vector was adopted to evaluate the DNA vaccine insertion and delivery. Three encapsulation protocols were tested to identify the most effective in internalizing the plasmid. The presence of plasmids inside the capsules was confirmed by fluorescence microscopy, and the encapsulation efficiency was calculated by the difference between the initial concentration of DNA used for insertion and the concentration of unencapsulated DNA contained in the supernatant. The capsules were subjected to different temperatures to evaluate their thermostability and were co-cultured with macrophages for phagocytosis analysis. HEK-293T cells were adopted to assess the cytotoxicity levels by MTT assay.

RESULTS

The microscopy results indicated that the macrophages successfully phagocytosed the capsules. Among the protocols tested for encapsulation, the one with 2% polyethylenimine for internalization showed the highest efficiency, with an encapsulation rate above 80%. However, the vaccine capsules obtained with the protocol that used 5% NaCl showed better thermal stability and encapsulation efficiency above 63% without induction of cell viability loss in HEK 293T.

CONCLUSIONS

We successfully described a vaccine delivery system using yeast capsules derived from , demonstrating its potential for DNA vaccine delivery for the first time. Additional studies will be needed to characterize and improve this delivery strategy.

摘要

背景/目的：DNA疫苗生产迅速且可适应不同病原体，但在稳定性和递送至细胞靶点方面面临巨大挑战。因此，有效的递送方法对这些疫苗的成功至关重要。在此，我们评估了源自酵母细胞壁的胶囊作为DNA疫苗递送系统的功效。

方法

从先前在YPD培养基中培养的酵母菌株GS115中提取胶囊。采用pVAX1表达载体评估DNA疫苗的插入和递送。测试了三种包封方案以确定内化质粒最有效的方案。通过荧光显微镜确认胶囊内质粒的存在，并通过用于插入的DNA初始浓度与上清液中未包封DNA浓度之间的差异计算包封效率。将胶囊置于不同温度下评估其热稳定性，并与巨噬细胞共培养以进行吞噬分析。采用HEK-293T细胞通过MTT法评估细胞毒性水平。