猪胸膜肺炎放线杆菌OmpD、LppB蛋白的原核表达及免疫原性分析

doi:10.11843/j.issn.0366-6964.2025.01.032

摘要/Abstract

摘要：

本研究旨在评价猪胸膜肺炎放线杆菌OmpD和LppB蛋白的免疫原性和保护效果。通过同源性分析，发现19个血清型的APP均含有ompD和lppB基因，氨基酸序列相似性分别为98.2%~100%和99.3%~100%。利用基因工程大肠杆菌表达纯化了1型菌株的rOmpD和rLppB蛋白，并进行了免疫原性分析。结果显示，两种蛋白均能有效表达，与猪康复血清反应明显。小鼠免疫rOmpD、rLppB蛋白后，ELISA抗体水平显著提高，对国内流行的1型、7型菌株攻毒保护率为70%~80%。进一步制备含rOmpD和rLppB的油乳剂，免疫仔猪后，ELISA抗体水平显著提高，对1型、7型菌株攻毒具有部分保护作用和交叉保护能力，但未能完全阻止感染和死亡。因此OmpD、LppB有作为APP亚单位疫苗开发候选抗原的潜力，本研究为进一步开发具有良好交叉保护效果的疫苗提供了数据支持。

关键词: 猪胸膜肺炎放线杆菌, OmpD, LppB, 免疫原性, 免疫效果

Abstract:

This study evaluated the immunogenicity and protective efficacy of OmpD and LppB proteins from Actinobacillus pleuropneumoniae. Homology analysis revealed that all 19 serotypes of APP have ompD and lppB genes, with amino acid sequence identities of 98.2%-100% and 99.3%-100%, respectively. Recombinant OmpD (rOmpD) and LppB (rLppB) proteins from serotype 1 were expressed and purified using engineered E. coli and subjected to immunogenicity analysis. The results showed that both proteins were effectively expressed and exhibited obvious reactivity with swine convalescent serum. Mice immunized with rOmpD or rLppB leaded to significantly elevated ELISA antibody titers and protection rates of 70%-80% against challenge with prevalent serotype 1 and 7 strains. Further, an oil emulsion containing rOmpD and rLppB was prepared, and immunization of piglets elicited significantly elevated ELISA antibody titers and conferred partial protection and cross-protection against serotype 1 and 7 challenge. However, complete protection against infection and death was not achieved. These findings suggest that OmpD and LppB are promising candidates for subunit vaccine development against Actinobacillus pleuropneumoniae, and provide valuable data for the further development of vaccines with cross-protective efficacy.

Key words: Actinobacillus pleuropneumoniae, OmpD, LppB, immunogenicity, efficacy

中图分类号:

S852.619

曾焱, 欧祥龙, 闫晓阳, 刘灿, 廖永洪. 猪胸膜肺炎放线杆菌OmpD、LppB蛋白的原核表达及免疫原性分析[J]. 畜牧兽医学报, 2025, 56(1): 343-352.

ZENG Yan, OU Xianglong, YAN Xiaoyang, LIU Can, LIAO Yonghong. Prokaryotic Expression and Immunogenicity Analysis of OmpD and LppB of Actinobacillus pleuropneumoniae[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(1): 343-352.

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参考文献 17

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扩增基因 Amplified gene	引物 Primer	引物序列(5′→3′) Primer sequence	片段长度/bp Segment length/bp	酶切位点 Cleavage sites
ompD	上游引物F	TCAGAATTCATGAAAAAATTCTTACTT	2 382	EcoRⅠ
ompD	下游引物R	CGACTCGAGGAACGAGCTACCGATACT	2 382	XhoⅠ
lppB	上游引物F	GATGAATTCATGAAGAAATCATTTTTT	1 239	EcoRⅠ
lppB	下游引物R	CAACTCGAGTTTTCTTGGTAAATAGC	1 239	XhoⅠ

组别 Groups	攻毒菌株 Challenge strain	小鼠数量 Number of mice	存活数量 Survival	死亡数量 Death	保护率/% Protection rate(%)
对照组 Control	Serotype 1 WZ2201	10	1	9	/
rOmpD疫苗组 rOmpD Vaccine		10	7	3	70
rLppB疫苗组 rLppB Vaccine		10	7	3	70
对照组 Control	Serotype 7 RC2103	10	2	8	/
rOmpD疫苗组 rOmpD Vaccine		10	7	3	70
rLppB疫苗组 rLppB Vaccine		10	8	2	80

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