表达猪流行性腹泻病毒S1基因重组罗伊氏黏液乳杆菌诱导仔猪特异性免疫应答的研究

doi:10.11843/j.issn.0366-6964.2025.09.036

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (9): 4581-4592.doi: 10.11843/j.issn.0366-6964.2025.09.036

表达猪流行性腹泻病毒S1基因重组罗伊氏黏液乳杆菌诱导仔猪特异性免疫应答的研究

国桂海¹(), 马茹梦¹, 尹方洁¹, 刘芯孜¹, 王梓³, 孟伟静⁴, 李佳璇¹^,², 崔文¹^,², 姜艳平¹^,², 唐丽杰¹^,², 赵海渊¹^,²^,*(), 王晓娜¹^,²^,*()

1. 东北农业大学动物医学学院, 哈尔滨 150030
2. 动物疾病防控技术与制剂创制省重点实验室, 哈尔滨 150030
3. 通辽市农牧科学研究所, 通辽 028000
4. 通辽市农牧业发展中心, 通辽 028000

收稿日期:2024-11-18 出版日期:2025-09-23 发布日期:2025-09-30
通讯作者: 赵海渊,王晓娜 E-mail:ggh2646859744@163.com;zhywxn1925@163.com;xiaonawang0319@163.com
作者简介:国桂海(2000-)，男，汉族，山东淄博人，硕士生，主要从事动物微生物学与免疫学研究，E-mail: ggh2646859744@163.com
基金资助:
国家自然科学基金青年项目(32102707)；黑龙江省自然科学基金联合引导项目(LH2021C043)

Study on the Expression of Recombinant Limosilactobacillus reuteri Expressing the S1 Gene of Porcine Epidemic Diarrhea Virus to Induce Specific Immune Responses in Piglets

GUO Guihai¹(), MA Rumeng¹, YIN Fangjie¹, LIU Xinzi¹, WANG Zi³, MENG Weijing⁴, LI Jiaxuan¹^,², CUI Wen¹^,², JIANG Yanping¹^,², TANG Lijie¹^,², ZHAO Haiyuan¹^,²^,*(), WANG Xiaona¹^,²^,*()

1. College of Veterinary Medicines, Northeast Agricultural University, Harbin 150030, China
2. Provincial Key Laboratory of Animal Disease Prevention and Control Technology and Preparation Development, Harbin 150030, China
3. Tongliao Institute of Animal Husbandry and Veterinary Science, Tongliao 028000, China
4. Tongliao Agricultural and Animal Husbandry Development Center, Tongliao 028000, China

Received:2024-11-18 Online:2025-09-23 Published:2025-09-30
Contact: ZHAO Haiyuan, WANG Xiaona E-mail:ggh2646859744@163.com;zhywxn1925@163.com;xiaonawang0319@163.com

摘要/Abstract

摘要：

旨在研究猪源罗伊氏黏液乳杆菌(Limosilactobacillus reuteri)作为活菌载体表达猪流行性腹泻病毒保护性抗原S1，口服免疫新生仔猪后进行免疫效果评价，以期研制安全高效的口服黏膜疫苗制剂。本研究构建表达PEDV S1的重组乳酸菌pPG-T7g10-S1/L. reuteri J 31，进行生长特性、重组质粒的遗传稳定性、体外IPEC-J2细胞黏附特性，并进行新生仔猪口服重组菌免疫效果的评估。结果显示，重组菌pPG-T7g10-S1/L. reuteri J 31相对于亲本株其生长特性未发生显著改变，质粒能够稳定遗传，在仔猪空肠具有较强的定植能力；新生仔猪口服免疫重组菌后，产生黏膜特异性SIgA抗体与血清特异性IgG抗体，且均具有中和PEDV的活性，能够诱导仔猪产生Th1、Th2和Th17型细胞免疫应答反应。综上所述，本研究成功构建重组乳酸菌pPG-T7g10-S1/L. reuteri J 31，口服免疫重组菌可诱导仔猪显著产生体液免疫、黏膜免疫和Th1、Th2和Th17型细胞免疫应答，为开发PEDV的口服疫苗候选制剂奠定了理论和实验基础。

关键词: 猪流行性腹泻, 猪源罗伊氏黏液乳杆菌, 口服疫苗

Abstract:

The aim of this study was to express the protective antigen S1 of porcine epidemic diarrhea virus (PEDV) using porcine Limosilactobacillus reuteri as a live bacterial vector, and to evaluate the immune effect after oral immunization of newborn piglets, in order to develop a safe and efficient oral mucosal vaccine preparation. In this study, the recombinant lactic acid strain pPG-T7g10-S1/L. ruteri J 31 expressing PEDV S1 was constructed. The growth characteristics, genetic stability of the recombinant plasmid, and adhesion characteristics of IPEC-J2 cells in vitro were evaluated. The results showed that the growth characteristics of the recombinant strain pPG-T7g10-S1/L. ruteri J 31 did not change significantly compared with the parental strain. The plasmid could be inherited stably and had a strong ability to colonize in the jejunum of piglets. After oral immunization of newborn piglets with the recombinant PEDV, the level of mucosa-specific SIgA antibody and serum specific IgG antibody were produced, and both of them had the activity of neutralizing PEDV, and could induce Th1, Th2 and Th17 types of cellular immune responses in piglets. In conclusion, in this study, the recombinant Limosilactobacillus reuteri pPG-T7g10-S1/L. ruteri J 31 was successfully constructed. Oral immunization with the pPG-T7G10-S1/L. ruteri J 31 could induce significant humoral immunity, mucosal immunity and Th1, Th2 and Th17 cellular immune responses in piglets. These results laid a theoretical and experimental foundation for the development of oral PEDV vaccine candidates.

Key words: porcine epidemic diarrhea, Limosilactobacillus reuteri, oral vaccine

中图分类号:

S859.797

国桂海, 马茹梦, 尹方洁, 刘芯孜, 王梓, 孟伟静, 李佳璇, 崔文, 姜艳平, 唐丽杰, 赵海渊, 王晓娜. 表达猪流行性腹泻病毒S1基因重组罗伊氏黏液乳杆菌诱导仔猪特异性免疫应答的研究[J]. 畜牧兽医学报, 2025, 56(9): 4581-4592.

GUO Guihai, MA Rumeng, YIN Fangjie, LIU Xinzi, WANG Zi, MENG Weijing, LI Jiaxuan, CUI Wen, JIANG Yanping, TANG Lijie, ZHAO Haiyuan, WANG Xiaona. Study on the Expression of Recombinant Limosilactobacillus reuteri Expressing the S1 Gene of Porcine Epidemic Diarrhea Virus to Induce Specific Immune Responses in Piglets[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4581-4592.

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菌株Strains	黏附率/% Rate of adhesion
L. reuteri J 31	39.24±1.11
pPG-T7g10-S1/L. reuteri J 31	41.18±1.44^*

组别Group	IgG	SIgA
PBS	0	0
pPG-T7g10-S1/L. reuteri J 31	1∶44.67 ^*	1∶17.78^*

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表达猪流行性腹泻病毒S1基因重组罗伊氏黏液乳杆菌诱导仔猪特异性免疫应答的研究

Study on the Expression of Recombinant Limosilactobacillus reuteri Expressing the S1 Gene of Porcine Epidemic Diarrhea Virus to Induce Specific Immune Responses in Piglets

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