Recombinant yeast-cell microcapsules carrying the DNA vaccine against enterotoxigenic Escherichia coli

被引：0

作者：

Zhao, Xiafang ^{[1
]}

Du, Lihong ^{[1
]}

Ma, Baoxia ^{[1
]}

Jia, Shaona ^{[1
]}

Liu, Yufei ^{[1
]}

Zhu, Yufei ^{[2
]}

Ma, Xiaotao ^{[2
]}

Yang, Xiaojun ^{[1
]}

Xu, Kun ^{[1
]}

机构：

[1] College of Animal Science and Technology, Northwest A&F University, Shaanxi, Yangling

[2] Shanxi Dayu Bioengineering Co., Ltd., Shanxi, Yuncheng

来源：

Shengwu Gongcheng Xuebao/Chinese Journal of Biotechnology | 2025年 / 41卷 / 06期

关键词：

DNA vaccines; enterotoxigenic Escherichia coli; intestinal immunity; intestinal microbiota; yeast-cell microcapsules;

D O I：

10.13345/j.cjb.240753

中图分类号：

学科分类号：

摘要：

The enterotoxigenic Escherichia coli (ETEC) infection is a major factor restricting the development of animal husbandry. However, the abuse of antibiotics will lead to the antibiotic residues and emergence of antibiotic-resistant bacteria. The existing vaccines face challenges in stimulating intestinal immunity, demonstrating limited prevention effects. Therefore, it is indispensable to develop a new vaccine that is safe and suitable as a feed additive to activate intestinal immunity. This study constructed yeast-cell microcapsules (YCM) carrying the DNA vaccine against ETEC by genetic engineering. Furthermore, animal experiments were carried out to explore the regulatory effects of feeding YCM on the intestinal immune system and intestinal microbiota. Saccharomyces cerevisiae was selected as the oral delivery vehicle (microcapsules) of the DNA vaccine. The codon-optimized nucleic acid sequence of K88, the main antigen of mammal-derived ETEC, was synthesized, and the yeast shuttle vector containing the corresponding DNA vaccine expression cassette was constructed by DNA recombination. The recombinant strain of YCM was prepared by transforming JMY1. Additionally, the characteristics of the YCM strain and its feasibility as an oral vaccine were comprehensively evaluated by the fluorescence reporter assay, gastrointestinal fluid tolerance assay, intestinal epithelial cell adhesion assay, intestinal retention assessment, antiserum detection, and intestinal microbiota detection. The experimental results showed that the DNA vaccine expression cassette was expressed in mammals, and the recombinant strain of YCM could tolerate up to 8 hours of gastrointestinal fluid digestion and had good adhesion to intestinal epithelial cells. The results of mouse feeding experiments indicated that the recombinant strain of YCM could stay in the intestinal tract for at least two weeks, and the DNA vaccine expression cassette carried by YCM entered the intestinal immune system and triggered an immune response to induce the production of specific antibodies. Moreover, feeding YCM recombinant bacteria also improved the abundance of gut microbiota in mice, demonstrating a positive effect in regulating intestinal flora. In summary, we prepared the recombinant strain of YCM carrying the DNA vaccine against ETEC and comprehensively evaluated its characteristics and feasibility as an oral vaccine. Feeding the recombinant YCM could induce specific immune responses and regulate intestinal microbiota. The findings provide a reference for the immunoprevention of ETEC-related animal diseases. ©2025 Chin J Biotech, All rights reserved.

引用

页码：2388 / 2404

页数：16

共 30 条

[1]

CHEN ZX, YONG TY, WEI ZH, ZHANG XQ, LI X, QIN JQ, LI JY, HU J, YANG XL, GAN L., Engineered probiotic-based personalized cancer vaccine potentiates antitumor immunity through initiating trained immunity, Advanced Science, 11, 3, (2024)

[2]

RILEY LW., Distinguishing pathovars from nonpathovars: Escherichia coli, Microbiology Spectrum, 8, 4, (2020)

[3]

SIITONEN A., Escherichia coli in fecal flora of healthy adults: serotypes, P and type 1C fimbriae, non-P mannose-resistant adhesins, and hemolytic activity, The Journal of Infectious Diseases, 166, 5, pp. 1058-1065, (1992)

[4]

KNUTTON S, SHAW R, PHILLIPS AD, SMITH HR, WILLSHAW GA, WATSON P, PRICE E., Phenotypic and genetic analysis of diarrhea-associated Escherichia coli isolated from children in the United Kingdom, Journal of Pediatric Gastroenterology and Nutrition, 33, 1, pp. 32-40, (2001)

[5]

TAN C, TANG XB, ZHANG X, DING Y, ZHAO ZQ, WU B, CAI XW, LIU ZF, HE QG, CHEN HC., Serotypes and virulence genes of extraintestinal pathogenic Escherichia coli isolates from diseased pigs in China, The Veterinary Journal, 192, 3, pp. 483-488, (2012)

[6]

FRANCIS DH, ERICKSON AK, GRANGE PA., K88 adhesins of enterotoxigenic Escherichia coli and their porcine enterocyte receptors, Advances in Experimental Medicine and Biology, 473, pp. 147-154, (1999)

[7]

BUTT S, GAGNON J, SALEH M., A protective role for glucagon-like peptide-2 in heat-stable enterotoxin b (STb)-induced L-cell toxicity, Endocrinology, 163, 4, (2022)

[8]

ZEGEYE ED, GOVASLI ML, SOMMERFELT H, PUNTERVOLL P., Development of an enterotoxigenic Escherichia coli vaccine based on the heat-stable toxin, Human Vaccines & Immunotherapeutics, 15, 6, pp. 1379-1388, (2019)

[9]

ZHANG HH, XU YP, ZHANG ZJ, YOU JS, YANG YY, LI XY., Protective immunity of a Multivalent Vaccine Candidate against piglet diarrhea caused by enterotoxigenic Escherichia coli (ETEC) in a pig model, Vaccine, 36, 5, pp. 723-728, (2018)

[10]

HOLMGREN J, CZERKINSKY C., Mucosal immunity and vaccines, Nature Medicine, 11, 4, pp. S45-853, (2005)

← 1 2 3 →