Genetic engineering employing MPB70 and its promoter enables efficient secretion and expression of foreign antigen in bacillus Calmette Guérin (BCG) Tokyo

IF 1.9 4区 医学 Q4 IMMUNOLOGY Microbiology and Immunology Pub Date : 2024-01-31 DOI:10.1111/1348-0421.13116
Atsuki Takeishi, Amina K. Shaban, Taichi Kakihana, Hayato Takihara, Shujiro Okuda, Hidekazu Osada, Desak Nyoman Surya Suameitria Dewi, Yuriko Ozeki, Yutaka Yoshida, Akihito Nishiyama, Yoshitaka Tateishi, Yuki Aizu, Yasushi Chuma, Kazuyo Onishi, Daisuke Hayashi, Saburo Yamamoto, Tetsu Mukai, Manabu Ato, Duong Huu Thai, Huynh Thi Thao Nhi, Tsuyoshi Shirai, Satoshi Shibata, Fumiko Obata, Jun Fujii, Seiya Yamayoshi, Maki Kiso, Sohkichi Matsumoto
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Abstract

Vaccination is an important factor in public health. The recombinant bacillus Calmette Guérin (rBCG) vaccine, which expresses foreign antigens, is expected to be a superior vaccine against infectious diseases. Here, we report a new recombination platform in which the BCG Tokyo strain is transformed with nucleotide sequences encoding foreign protein fused with the MPB70 immunogenic protein precursor. By RNA-sequencing, mpb70 was found to be the most transcribed among all known genes of BCG Tokyo. Small oligopeptide, namely, polyhistidine tag, was able to be expressed in and secreted from rBCG through a process in which polyhistidine tag fused with intact MPB70 were transcribed by an mpb70 promoter. This methodology was applied to develop an rBCG expressing the receptor binding domain (RBD) of severe acute respiratory syndrome coronavirus 2. Immunoblotting images and mass spectrometry data showed that RBD was also secreted from rBCG. Sera from mice vaccinated with the rBCG showed a tendency of weak neutralizing capacity. The secretion was retained even after a freeze-drying process. The freeze-dried rBCG was administered to and recovered from mice. Recovered rBCG kept secreting RBD. Collectively, our recombination platform offers stable secretion of foreign antigens and can be applied to the development of practical rBCGs.

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利用 MPB70 及其启动子的基因工程技术,可以在东京卡介苗(BCG)中高效分泌和表达外来抗原。
疫苗接种是公共卫生的一个重要因素。表达外来抗原的重组卡介苗(rBCG)有望成为预防传染病的优质疫苗。在这里,我们报告了一种新的重组平台,在该平台中,卡介苗东京株被转化为编码与 MPB70 免疫原蛋白前体融合的外来蛋白的核苷酸序列。通过 RNA 测序,我们发现 mpb70 是东京卡介苗所有已知基因中转录最多的基因。通过聚组氨酸标签与完整的 MPB70 相融合,在 mpb70 启动子的转录过程中,小的寡肽(即聚组氨酸标签)能够在 rBCG 中表达并从 rBCG 中分泌出来。这种方法被用于开发表达严重急性呼吸系统综合征冠状病毒 2 受体结合域(RBD)的 rBCG。免疫印迹图像和质谱数据显示,RBD 也从 rBCG 中分泌。接种了 rBCG 疫苗的小鼠血清显示出弱中和能力的趋势。即使经过冷冻干燥过程,这种分泌物仍能保留下来。将冻干的 rBCG 给小鼠注射并从小鼠体内回收。回收的 rBCG 继续分泌 RBD。总之,我们的重组平台能稳定分泌外来抗原,可用于开发实用的 rBCG。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Microbiology and Immunology
Microbiology and Immunology 医学-免疫学
CiteScore
5.20
自引率
3.80%
发文量
78
审稿时长
1 months
期刊介绍: Microbiology and Immunology is published in association with Japanese Society for Bacteriology, Japanese Society for Virology, and Japanese Society for Host Defense Research. It is peer-reviewed publication that provides insight into the study of microbes and the host immune, biological and physiological responses. Fields covered by Microbiology and Immunology include:Bacteriology|Virology|Immunology|pathogenic infections in human, animals and plants|pathogenicity and virulence factors such as microbial toxins and cell-surface components|factors involved in host defense, inflammation, development of vaccines|antimicrobial agents and drug resistance of microbes|genomics and proteomics.
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