表达非洲猪瘟病毒基因B602L-B646L的重组病毒免疫原性的初步分析

doi:10.11843/j.issn.0366-6964.2025.06.025

摘要/Abstract

摘要：

本研究旨在分析表达非洲猪瘟病毒基因B602L-B646L的重组腺病毒和杆状病毒的免疫原性。联合CpG佐剂注射3 d后或单独免疫rAd-F-Hsp70 21 d后用rBac-F加强免疫，或以rBac-F为初免21 d后用rAd-F-Hsp70进行加强免疫，免疫剂量1×10⁸·只^-1。在初免21和42 d后采集血清检测针对ASFV pB602L和p72蛋白的抗体水平。检测初免42 d后的由抗原刺激后外周血单核细胞(PBMCs)上清中干扰素-γ(IFN-γ)和白细胞介素-4(IL-4)的含量。结果显示：自免疫后第21天起，所有免疫猪的血清中均能检测到特异性IgG抗体，且在加强免疫后，抗体水平显著上升。CpG联合rAd-F-Hsp70/rBac-F免疫组血清中，pB602L和p72抗体水平均显著高于rAd-F-Hsp70/rBac-F(P < 0.01)和rBac-F/rAd-F-Hsp70(P < 0.01)，其中rAd-F-Hsp70/rBac-F免疫组特异性抗体水平也显著高于rBac-F/rAd-F-Hsp70(P < 0.01)。用ASFV pB602L和p72重组抗原刺激PBMCs后，CpG佐剂联合rAd-F-Hsp70/rBac-F免疫组细胞上清中IFN-γ和IL-4含量显著高于其他免疫组(P < 0.01)，表明以rAd-F-Hsp70作为首免载体优于rBac-F以及CpG佐剂显著提高体液免疫和细胞免疫效果。综上，以rAd-F-Hsp70作为初免疫苗和以rBac-F作为加强疫苗，同时联合使用CpG佐剂显著增强了针对pB602L和p72特异性抗体和细胞因子水平，为进一步优化非洲猪瘟病毒相关疫苗策略提供了数据支持和参考依据。

关键词: 非洲猪瘟病毒, 重组腺病毒, 重组杆状病毒, CpG分子

Abstract:

This study aimed to analyze the immunogenicity of recombinant adenoviruses and baculoviruses expressing African Swine Fever Virus (ASFV) genes B602L-B646L. Piglets were immunized with rAd-F-Hsp70, followed by a booster with rBac-F either 21 days later or 3 days after co-administration with the CpG adjuvant. Alternatively, the immunization strategy involved rBac-F as the initial vaccine, followed by a booster with rAd-F-Hsp70 after 21 days. Each piglet received a dose of 1×10⁸ PFU. Serum was collected on days 21 and 42 post-primary immunization to measure antibody levels against ASFV pB602L and p72 proteins. Additionally, the levels of interferon-γ (IFN-γ) and interleukin-4 (IL-4) in the supernatants of antigen-stimulated peripheral blood mononuclear cells (PBMCs) were assessed on day 42 post-primary immunization. Results were as follows: Specific IgG antibodies were detected in the serum of all immunized pigs from day 21 onwards, with antibody levels significantly increasing after the booster immunization. The CpG adjuvant combined with rAd-F-Hsp70/rBac-F group showed significantly higher levels of pB602L and p72 antibodies compared to both the rAd-F-Hsp70/rBac-F (P < 0.01) and rBac-F/rAd-F-Hsp70 groups (P < 0.01). Among these, the rAd-F-Hsp70/rBac-F group exhibited significantly higher specific antibody levels compared to the rBac-F/rAd-F-Hsp70 group (P < 0.01). After stimulation of PBMCs with ASFV pB602L and p72 recombinant antigens, the supernatants from the CpG adjuvant combined with the rAd-F-Hsp70/rBac-F group contained significantly higher levels of IFN-γ and IL-4 compared to the other immunized groups (P < 0.01). This indicates that using rAd-F-Hsp70 as the primary immunization vector, combined with CpG adjuvant, significantly enhances both humoral and cellular immune responses. Using rAd-F-Hsp70 as the primary vaccine and rBac-F as the booster, along with the CpG adjuvant, significantly enhances the levels of specific antibodies and cytokines against pB602L and p72. These findings provide data to optimize ASFV vaccine strategies for future development.

Key words: African swine fever virus, recombinant adenovirus, recombinant baculovirus, CpG

中图分类号:

S852.659.1

卢会鹏, 曹世诺, 吴植, 陈长春, 陈文玉, 成玉婷, 周晓慧, 孙怀昌, 朱善元. 表达非洲猪瘟病毒基因B602L-B646L的重组病毒免疫原性的初步分析[J]. 畜牧兽医学报, 2025, 56(6): 2816-2825.

LU Huipeng, CAO Shinuo, WU Zhi, CHEN Changchun, CHEN Wenyu, CHENG Yuting, ZHOU Xiaohui, SUN Huaichang, ZHU Shanyuan. The Preliminary Analysis of the Immunogenicity of Recombinant Viruses Expressing African Swine Fever Virus Genes B602L-B646L[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(6): 2816-2825.

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引物Primer	引物序列(5′→3′) Sequence	酶切位点Enzyme cleavage site
B646L For	TAAGAAGGAGATATACATATGAGCAACATCAAGAACGTC	NdeⅠ
B646L Rev	GTGGTGGTGGTGGTGCTCGAGGTCGGTGATGGGGGTGAT	XhoⅠ
B602L For	TAAGAAGGAGATATACATATGGACTCTAGTAATCAGCAGA	NdeⅠ
B602L Rev	GTGGTGGTGGTGGTGCTCGAGCAGCTCGGCCTTGGTGTA	XhoⅠ

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