Dataset Information

Preparation and Identification of a Monoclonal Antibody against the Pseudorabies Virus gE Glycoprotein through a Novel Strategy.

ABSTRACT: Since 2011, pseudorabies virus (PRV) has recurred in several vaccinated pig farms in China. PRV variants with high virulence were found to be the main cause of the outbreaks. In the face of the PRV epidemic, detection of the wild strain is as important as vaccine immunization, so we hoped to achieve differential diagnosis of PRV by obtaining a monoclonal antibody (mAB) that could be used to identify the wild strain. In this study, we used a novel immunization and screening strategy to prepare an mAB and obtained mAB 1H5 against the gE glycoprotein. An immunofluorescence assay (IFA) revealed that this mAB was specific to both classic and variant strains of PRV. Subsequently, we further identified the linear epitopes of B cells recognized using the mAB. The mAB 1H5 bound at ⁶⁷RRAG⁷⁰, which is a novel epitope and is conserved in almost all PRV strains. These findings provide novel insight into the structure and function of PRV proteins, the analysis of antigenic epitope characteristics, and the establishment of antigen or antibody detection methods.

SUBMITTER: Guo Z

PROVIDER: S-EPMC9968200 | biostudies-literature | 2023 Feb

REPOSITORIES: biostudies-literature

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Preparation and Identification of a Monoclonal Antibody against the Pseudorabies Virus gE Glycoprotein through a Novel Strategy.

Guo Zhenyang Z Zhang Siyu S Xu Hu H Li Wansheng W Li Chao C Zhao Jing J Gong Bangjun B Sun Qi Q Xiang Lirun L Zhao Hongyuan H Wang Qian Q Zhou Guohui G Tang Yandong Y An Tongqing T Cai Xuehui X Tian Zhijun Z Zhang Hongliang H Peng Jinmei J

Veterinary sciences 20230209 2

Since 2011, pseudorabies virus (PRV) has recurred in several vaccinated pig farms in China. PRV variants with high virulence were found to be the main cause of the outbreaks. In the face of the PRV epidemic, detection of the wild strain is as important as vaccine immunization, so we hoped to achieve differential diagnosis of PRV by obtaining a monoclonal antibody (mAB) that could be used to identify the wild strain. In this study, we used a novel immunization and screening strategy to prepare an ...[more]

PMID: 36851437

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Project description:BackgroundSince the end of 2011 an outbreak of pseudorabies affected Chinese pig herds that had been vaccinated with the commercial vaccine made of Bartha K61 strain. It is now clear that the outbreak was caused by an emergent PRV variant. Even though vaccines made of PRV Bartha K61 strain can confer certain cross protection against PRV variants based on experimental data, less than optimal clinical protection and virus shedding reduction were observed, making the control or eradication of this disease difficult.ResultsAn infectious clone of PRV AH02LA strain was constructed to generate a gE deletion mutant PRV(LA-AB) strain. PRV(LA-AB) strain can reach a titer of 108.43 TCID50 /mL (50% tissue culture infectious dose) on BHK-21 cells. To evaluate the efficiency of the inactivated vaccine made of PRV(LA-AB) strain, thirty 3-week-old PRV-negative piglets were divided randomly into six groups for vaccination and challenge test. All five piglets in the challenge control showed typical clinical symptoms of pseudorabies post challenge. Sneezing and nasal discharge were observed in four and three piglets in groups C(vaccinated with inactivated PRV Bartha K61 strain vaccine) and D(vaccinated with live PRV Bartha K61 strain vaccine) respectively. In contrast, piglets in both groups A(vaccinated with inactivated PRV LA-AB strain vaccine) and B(vaccinated with inactivated PRV LA-AB strain vaccine with adjuvant) presented mild or no clinical symptoms. Moreover, viral titers detected via nasal swabs were approximately 100 times lower in group B than in the challenge control, and the duration of virus shedding (3-4 days) was shorter than in either the challenge control (5-10 days) or groups C and D (5-6 days).ConclusionsThe infectious clone constructed in this study harbors the whole genome of the PRV variant AH02LA strain. The gE deletion mutant PRV(LA-AB)strain generated from PRV AH02LA strain can reach a high titer on BHK-21 cells. An inactivated vaccine of PRV LA-AB provides clinical protection and significantly reduces virus shedding post challenge, especially if accompanied by the adjuvant CVC1302. While Inactivated or live vaccines made of PRV Barth K61 strain can provide only partial protection in this test.

Dataset Information

Preparation and Identification of a Monoclonal Antibody against the Pseudorabies Virus gE Glycoprotein through a Novel Strategy.

Publications

Preparation and Identification of a Monoclonal Antibody against the Pseudorabies Virus gE Glycoprotein through a Novel Strategy.

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