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

被引：0

作者：

Takeishi, Atsuki ^{[1
]}

Shaban, Amina K. ^{[1
]}

Kakihana, Taichi ^{[2
]}

Takihara, Hayato ^{[3
]}

Okuda, Shujiro ^{[3
]}

Osada, Hidekazu ^{[1
,4
]}

Dewi, Desak Nyoman Surya Suameitria ^{[1
,5
]}

Ozeki, Yuriko ^{[1
]}

Yoshida, Yutaka ^{[1
]}

Nishiyama, Akihito ^{[1
]}

Tateishi, Yoshitaka ^{[1
]}

Aizu, Yuki ^{[6
]}

Chuma, Yasushi ^{[6
]}

Onishi, Kazuyo ^{[6
]}

Hayashi, Daisuke ^{[6
]}

Yamamoto, Saburo ^{[6
,7
]}

Mukai, Tetsu ^{[7
]}

Ato, Manabu ^{[7
]}

Thai, Duong Huu ^{[8
]}

Nhi, Huynh Thi Thao ^{[9
]}

Shirai, Tsuyoshi ^{[10
]}

Shibata, Satoshi ^{[11
]}

Obata, Fumiko ^{[11
]}

Fujii, Jun ^{[11
]}

Yamayoshi, Seiya ^{[12
]}

Kiso, Maki ^{[12
]}

Matsumoto, Sohkichi ^{[1
,13
,14
]}

机构：

[1] Niigata Univ, Sch Med, Dept Bacteriol, Niigata, Japan

[2] Niigata Univ, Sch Med, Dept Immunol, Niigata, Japan

[3] Niigata Univ, Med AI Ctr, Sch Med, Niigata, Japan

[4] NIPPON ZENYAKU KOGYO CO LTD, Fukushima, Japan

[5] Univ Ciputra, Microbiol, Surabaya, Indonesia

[6] Div Res & Dev, Japan BCG Lab, Tokyo, Japan

[7] Natl Inst Infect Dis, Leprosy Res Ctr, Dept Mycobacteriol, Tokyo, Japan

[8] Inst Vaccines & Med Biol, Nha Trang, Vietnam

[9] Inst Vaccines & Med Biol, Nha Trang, Vietnam

[10] Nagahama Inst Biosci & Technol, Dept Biosci, Nagahama, Shiga, Japan

[11] Tottori Univ, Fac Med, Dept Microbiol & Immunol, Div Bacteriol, Tottori, Japan

[12] Univ Tokyo, Inst Med Sci, Tokyo, Japan

[13] Univ Airlangga, Fac Med, Dept Med Microbiol, Surabaya, Indonesia

[14] Hokkaido Univ, Inst Vaccine Res & Dev, Div Res Aids, Sapporo, Hokkaido, Japan

来源：

MICROBIOLOGY AND IMMUNOLOGY | 2024年 / 68卷 / 04期

关键词：

BCG; freeze-drying; MPB70; recombinant BCG; RNA-seq; MYCOBACTERIUM-BOVIS BCG; HOMOLOGOUS RECOMBINATION; ESCHERICHIA-COLI; IMMUNE-RESPONSE; TUBERCULOSIS; PROTEIN; SUBSTRAINS; STRAINS; MPB83; IDENTIFICATION;

D O I：

10.1111/1348-0421.13116

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Vaccination is an important factor in public health. The recombinant bacillus Calmette Guerin (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.

引用

页码：130 / 147

页数：18