表达非洲猪瘟pp62与Hsp70蛋白重组腺病毒对小鼠的免疫原性分析

doi:10.11843/j.issn.0366-6964.2025.06.044

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (6): 3027-3031.doi: 10.11843/j.issn.0366-6964.2025.06.044

表达非洲猪瘟pp62与Hsp70蛋白重组腺病毒对小鼠的免疫原性分析

陈长春(), 吴植, 任冠宇, 陈文玉, 曹世诺, 朱睿, 张力, 成玉婷, 朱善元, 卢会鹏*()

江苏农牧科技职业学院，泰州 225300

收稿日期:2024-07-26 出版日期:2025-06-23 发布日期:2025-06-25
通讯作者: 卢会鹏 E-mail:chenchangchun66@126.com;luhuipeng@jsahvc.edu.cn
作者简介:陈长春(1975-)，男，安徽金寨人，硕士，主要从事病毒学研究，E-mail：chenchangchun66@126.com
基金资助:
江苏农牧科技职业学院自然科学基金储备项目(NSF2022CB22)；江苏省重点研发计划(现代农业)(BE2020407)；江苏农牧科技职业学院科技创新团队资助项目(NSF2023TC01)

Immunogenicity Analysis of Recombinant Adenovirus Expressing African Swine Fever Virus (ASFV) pp62 and Hsp70 Proteins in Mice

CHEN Changchun(), WU Zhi, REN Guanyu, CHEN Wenyu, CAO Shinuo, ZHU Rui, ZHANG Li, CHENG Yuting, ZHU Shanyuan, LU Huipeng*()

Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China

Received:2024-07-26 Online:2025-06-23 Published:2025-06-25
Contact: LU Huipeng E-mail:chenchangchun66@126.com;luhuipeng@jsahvc.edu.cn

摘要/Abstract

摘要：

本研究旨在开发一种针对非洲猪瘟病毒(African swine fever virus, ASFV)的重组腺病毒疫苗，为ASFV疫苗的临床应用和研发提供科学依据。利用pAdEasy-1系统构建融合表达ASFV CP530R基因和分支结核杆菌热休克蛋白70(Hsp70)的重组腺病毒(rAd-CP530R-Hsp70)。通过间接免疫荧光和Western blot验证目的蛋白的表达。将rAd-CP530R-Hsp70重组腺病毒免疫小鼠，检测小鼠体内产生的抗体和细胞因子水平。结果显示：重组腺病毒成功诱导小鼠产生针对pp62的特异性抗体，免疫后35 d抗体滴度达到1.33。免疫后小鼠脾细胞培养上清中IL-2、IL-4和IFN-γ水平显著升高，显示该疫苗能够有效激活体液和细胞免疫应答。构建的重组腺病毒疫苗可有效诱导小鼠产生免疫应答，为非洲猪瘟疫苗的研发提供数据支持。

关键词: 非洲猪瘟病毒, CP530R, 重组腺病毒, Hsp

Abstract:

The aim of this study was to develop a recombinant adenovirus vaccine against African swine fever virus (ASFV) to provide a scientific basis for the clinical application and development of ASFV vaccines. We constructed a recombinant adenovirus (rAd-CP530R-Hsp70) expressing the fusion of ASFV CP530R gene and Mycobacterium tuberculosis heat shock protein 70 (Hsp70) using the pAdEasy-1 system. The expression of the target protein was verified by indirect immunofluorescence and Western blot. Mice were immunized with the rAd-CP530R-Hsp70 recombinant adenovirus, and the levels of antibodies and cytokines produced in the mice were detected. Results were as follows: The recombinant adenovirus successfully induced mice to produce specific antibodies against pp62, with antibody titers reaching 1.33 at 35 days post-immunization. The levels of IL-2, IL-4, and IFN-γ in the supernatant of mouse splenocyte cultures were significantly increased after immunization, indicating that the vaccine could effectively activate both humoral and cellular immune responses. The constructed recombinant adenovirus vaccine can effectively induce an immune response in mice, providing data support for the development of African swine fever vaccines.

Key words: African swine fever virus, CP530R, recombinant adenovirus, Hsp70

中图分类号:

S852.659.1

陈长春, 吴植, 任冠宇, 陈文玉, 曹世诺, 朱睿, 张力, 成玉婷, 朱善元, 卢会鹏. 表达非洲猪瘟pp62与Hsp70蛋白重组腺病毒对小鼠的免疫原性分析[J]. 畜牧兽医学报, 2025, 56(6): 3027-3031.

CHEN Changchun, WU Zhi, REN Guanyu, CHEN Wenyu, CAO Shinuo, ZHU Rui, ZHANG Li, CHENG Yuting, ZHU Shanyuan, LU Huipeng. Immunogenicity Analysis of Recombinant Adenovirus Expressing African Swine Fever Virus (ASFV) pp62 and Hsp70 Proteins in Mice[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 3027-3031.

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载体 Vector	引物名称 Primer designation	引物序列(5′→3′) Primer sequence	酶切位点 Restriction site
pShuttle-CMV	CP530R-F1	ACCGTCGACGCCACCATGCCCAGCAACATGAAG	SalⅠ
	CP530R-R1	AGGCTCGAGCTCCTGCAGCAGCTTGAG	XhoⅠ
pET28a	CP530R-F2	CGCGGATCCTACAAGACCCACGGCGAC	BamHⅠ
	CP530R-R2	CTCGAATTCTCCTGCAGCAGCTTGAGG	EcoRⅠ

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表达非洲猪瘟pp62与Hsp70蛋白重组腺病毒对小鼠的免疫原性分析

Immunogenicity Analysis of Recombinant Adenovirus Expressing African Swine Fever Virus (ASFV) pp62 and Hsp70 Proteins in Mice

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