A highly pathogenic porcine reproductive and respiratory syndrome virus candidate vaccine based on Japanese encephalitis virus replicon system

Hu, Ping; Chen, Xiaoming; Huang, Lihong; Liu, Shukai; Zang, Fuyu; Xing, Jinchao; Zhang, Youyue; Liang, Jiaqi; Zhang, Guihong; Liao, Ming; Qi, Wenbao

doi:10.7717/peerj.3514

Cited by 3 publications

(2 citation statements)

References 36 publications

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“…The membrane was washed three times with TBST. Then, the membrane was analyzed by using an Odyssey Infrared Imaging System (LI-COR Biosciences, USA) (Xie et al, 2014;Hu et al, 2017). The samples were stored at −80°C.…”

Section: Western Blotmentioning

confidence: 99%

Serine 105 and 120 are important phosphorylation sites for porcine reproductive and respiratory syndrome virus N protein function

Yao

Xing

et al. 2018

Veterinary Microbiology

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The nucleocapsid (N) protein is the most abundant protein of porcine reproductive and respiratory syndrome virus (PRRSV). It has been shown to be multiphosphorylated. However, the phosphorylation sites are still unknown. In this study, we used liquid chromatography tandem mass spectrometry (LC-MS/MS) to analyze the phosphorylation sites of N protein expressed in Sf9 cells. The results showed that N protein contains two phosphorylation sites. Since N protein can regulate IL-10, which may facilitate PRRSV replication, we constructed four plasmids (pCA-XH-GD, pCA-A105, pCA-A120 and pCA-A105-120) and transfected them into Pig alveolar macrophages (PAMs,3D4/2). The qPCR results showed that mutations at residues 105 and 120 were associated with down-regulation of the IL-10 mRNA level, potentially decreasing the viral growth ability. Then, we mutated the phosphorylation sites (S105A and S120A) and rescued three mutated viruses, namely, A105, A120 and A105-120. Compared with wild-type virus titers, the titers of the mutated viruses at 48 hpi were significantly decreased. The Nsp(non-structural protein) 9 qPCR results were consistent with the multistep growth kinetics results. The infected PAMs (primary PAMs) results were similar with Marc-145.The findings indicated that the mutations impaired the viral replication ability. The confocal microscopy results suggested that mutations to residues 105 and 120 did not affect N protein distribution. Whether the mutations affected other functions of N protein and what the underlying mechanisms are need further investigation. In conclusion, our results show that residues 105 and 120 are phosphorylation sites and are important for N protein function and for viral replication ability.

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Section: Western Blotmentioning

confidence: 99%

Serine 105 and 120 are important phosphorylation sites for porcine reproductive and respiratory syndrome virus N protein function

Yao

Xing

et al. 2018

Veterinary Microbiology

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“…The results showed that recombinant PRRSV rHuN4-F112-Δ508-532 could be used as a potential marker vaccine for PRRS. Hu et al [ 100 ] constructed a live Japanese encephalitis virus (JEV) vaccine, SA14-14-2, with a CMV promoter and inserted PRRSV GP5/M into the deletion site of the replicon, based on JEV DNA, to develop the Balb/c chimeric replicon vaccine candidates pJEV-REP-G-2A-M-IRES and pJEV-REP-G-2A-M for animal immunoassays. ELISA data analysis showed that GP5/M replicons induce better immune responses; therefore, GP5/M JEV DNA-based replicons could be further developed into a new and safe PRRSV vaccine candidate.…”

Section: Gene-deletion Vaccinesmentioning

confidence: 99%

Research Progress on the Development of Porcine Reproductive and Respiratory Syndrome Vaccines

Zhang,

Luo,

et al. 2023

Veterinary Sciences

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Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious disease in the pig industry, but its pathogenesis is not yet fully understood. The disease is caused by the PRRS virus (PRRSV), which primarily infects porcine alveolar macrophages and disrupts the immune system. Unfortunately, there is no specific drug to cure PRRS, so vaccination is crucial for controlling the disease. There are various types of single and combined vaccines available, including live, inactivated, subunit, DNA, and vector vaccines. Among them, live vaccines provide better protection, but cross-protection is weak. Inactivated vaccines are safe but have poor immune efficacy. Subunit vaccines can be used in the third trimester of pregnancy, and DNA vaccines can enhance the protective effect of live vaccines. However, vector vaccines only confer partial protection and have not been widely used in practice. A PRRS vaccine that meets new-generation international standards is still needed. This manuscript provides a comprehensive review of the advantages, disadvantages, and applicability of live-attenuated, inactivated, subunit, live vector, DNA, gene-deletion, synthetic peptide, virus-like particle, and other types of vaccines for the prevention and control of PRRS. The aim is to provide a theoretical basis for vaccine research and development.

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Immune responses and protective effects against Japanese encephalitis induced by a DNA vaccine encoding the prM/E proteins of the attenuated SA14-14-2 strain

Zheng

Wang

et al. 2020

Infection, Genetics and Evolution

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A highly pathogenic porcine reproductive and respiratory syndrome virus candidate vaccine based on Japanese encephalitis virus replicon system

Cited by 3 publications

References 36 publications

Serine 105 and 120 are important phosphorylation sites for porcine reproductive and respiratory syndrome virus N protein function

Serine 105 and 120 are important phosphorylation sites for porcine reproductive and respiratory syndrome virus N protein function

Research Progress on the Development of Porcine Reproductive and Respiratory Syndrome Vaccines

Immune responses and protective effects against Japanese encephalitis induced by a DNA vaccine encoding the prM/E proteins of the attenuated SA14-14-2 strain

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