基于乳酸乳球菌的表达HIV gp120 DNA疫苗的免疫学分析

Gregers J Gram, Anders Fomsgaard, Mette Thorn, Søren M Madsen, Jacob Glenting
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引用次数: 17

摘要

由于效率的原因,大肠杆菌今天被用作生产质粒DNA疫苗的微生物工厂。为了避免目前使用的质粒系统产生有害的抗生素耐药基因和内毒素,我们开发了一种基于食品级乳酸乳球菌和不含抗生素耐药基因的质粒的系统。我们将乳酸菌系统与大肠杆菌中的传统系统进行了比较,使用相同的疫苗结构编码HIV-1的gp120。转染研究表明,两种载体系统的gp120表达水平相当。乳杆菌载体肌内免疫小鼠产生的gp120抗体滴度与大肠杆菌载体免疫小鼠的gp120抗体滴度相当。相反,L. lactis载体对细胞溶解反应的诱导作用较低。当使用大肠杆菌而不是乳杆菌载体时,在质粒中包含CpG基序更能提高t细胞的活性。这可能是由于载体主干的DNA含量不同。有趣的是,刺激脾细胞显示乳酸菌质粒具有较高的佐剂作用。研究结果表明,该乳酸菌质粒体系可作为一种新的替代DNA疫苗体系,具有较好的安全性。相似的基因表达单元、不同的载体骨架和不同的生产宿主具有不同的免疫诱导特性,这为沉默质粒骨架的佐剂作用提供了信息。结果还表明,质粒DNA的体外佐剂性与其诱导小鼠细胞和体液免疫反应的能力之间并不是直接相关的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Immunological analysis of a Lactococcus lactis-based DNA vaccine expressing HIV gp120.

Immunological analysis of a Lactococcus lactis-based DNA vaccine expressing HIV gp120.

Immunological analysis of a Lactococcus lactis-based DNA vaccine expressing HIV gp120.

Immunological analysis of a Lactococcus lactis-based DNA vaccine expressing HIV gp120.

For reasons of efficiency Escherichia coli is used today as the microbial factory for production of plasmid DNA vaccines. To avoid hazardous antibiotic resistance genes and endotoxins from plasmid systems used nowadays, we have developed a system based on the food-grade Lactococcus lactis and a plasmid without antibiotic resistance genes. We compared the L. lactis system to a traditional one in E. coli using identical vaccine constructs encoding the gp120 of HIV-1. Transfection studies showed comparable gp120 expression levels using both vector systems. Intramuscular immunization of mice with L. lactis vectors developed comparable gp120 antibody titers as mice receiving E. coli vectors. In contrast, the induction of the cytolytic response was lower using the L. lactis vector. Inclusion of CpG motifs in the plasmids increased T-cell activation more when the E. coli rather than the L. lactis vector was used. This could be due to the different DNA content of the vector backbones. Interestingly, stimulation of splenocytes showed higher adjuvant effect of the L. lactis plasmid. The study suggests the developed L. lactis plasmid system as new alternative DNA vaccine system with improved safety features. The different immune inducing properties using similar gene expression units, but different vector backbones and production hosts give information of the adjuvant role of the silent plasmid backbone. The results also show that correlation between the in vitro adjuvanticity of plasmid DNA and its capacity to induce cellular and humoral immune responses in mice is not straight forward.

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