title: HP-PRRSV is attenuated by de-optimization of codon pair bias in its RNA-dependent RNA polymerase nsp9 gene.
abstract: There is an urgent need to develop new vaccines against highly pathogenic PRRS virus (HP-PRRSV) variant in China. The actual use of each codon pairs is more or less frequent than that of the statistical prediction and codon pair bias (CPB) usage affects gene translation. We "shuffled" the existing codons in HP-PRRSV genes GP5, M, nsp2 and nsp9, so that the CPB of these genes could be more negative. De-optimization of nsp9, the RNA-dependent RNA polymerase, significantly decreased PRRSV replication in porcine alveolar macrophages (PAMs). In vitro study showed that HV-nsp9(min) and HV-nsp29(min) were remarkably attenuated in PAMs, and inoculation of pigs with 2 ml⁎10(5.0) TCID50/ml of HV-nsp9(min) or HV-nsp29(min) did not cause PRRS. Importantly, pigs immunized with HV-nsp29(min) were fully protected against different HP-PRRSV strains׳ lethal challenges. Our results imply that the CPB de-optimized HV-nsp29(min) has the potential to be used as a live vaccine candidate against HP-PRRSV.
authors.affiliation: China Academy of Medicine Sciences, Peking Union Medical College, Institute of Medicinal Plant Development, Beijing 100193, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Agrobiotechnology, Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Lianghai
authors.initials: L
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Agrobiotechnology, Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Huang
authors.affiliation: The Hamner Institutes for Health Sciences, Research Triangle Park, NC 27709-2137, USA.
authors.forename: Longlong
authors.initials: L
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Agrobiotechnology, Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Xue-Kun
authors.initials: XK
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Agrobiotechnology, Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Zhibin
authors.initials: Z
authors.lastname: Yu
authors.affiliation: State Key Laboratory of Agrobiotechnology, Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China.
authors.forename: Yihao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Beijing Senkang Biotech Development Co., Ltd, Beijing 101400, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Agrobiotechnology, Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China; Department of Microbiology and Immunology, College of Biological Sciences, China Agricultural University, Beijing 100193, China. Electronic address: whfeng@cau.edu.cn.
authors.forename: Wen-Hai
authors.initials: WH
authors.lastname: Feng
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis; D003062:Codon / Q000737:chemistry* / Q000276:immunology; D005818:Genetic Engineering; D007114:Immunization; D016676:Macrophages, Alveolar / Q000187:drug effects* / Q000276:immunology / Q000821:virology; D019318:Porcine Reproductive and Respiratory Syndrome / Q000276:immunology / Q000401:mortality / Q000517:prevention & control* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000187:drug effects* / Q000235:genetics / Q000276:immunology; D014176:Protein Biosynthesis; D012324:RNA-Dependent RNA Polymerase / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D016019:Survival Analysis; D013552:Swine; D014613:Vaccines, Attenuated; D019562:Viral Load; D014764:Viral Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D003062:Codon; D014613:Vaccines, Attenuated; D014764:Viral Proteins; D014765:Viral Vaccines; D012324:RNA-Dependent RNA Polymerase
keywords: Attenuation; Codon pair bias; HP-PRRSV; RNA dependent RNA polymerase; Vaccine