PMID- 29442228 OWN - NLM STAT- MEDLINE DCOM- 20180529 LR - 20191210 IS - 1432-8798 (Electronic) IS - 0304-8608 (Linking) VI - 163 IP - 6 DP - 2018 Jun TI - Effect of an 88-amino-acid deletion in nsp2 of porcine reproductive and respiratory syndrome virus on virus replication and cytokine responses in vitro. PG - 1489-1501 LID - 10.1007/s00705-018-3760-7 [doi] AB - Previously, a spontaneous 88-amino-acid (aa) deletion in nsp2 was associated with cell-adaptation of porcine reproductive and respiratory syndrome virus (PRRSV) strain JXM100, which arose during passaging of the highly pathogenic PRRSV (HP-PRRSV) strain JX143 in MARC-145 cells. Here, to elucidate the biological role of this deletion, we specifically deleted the region of a cDNA clone of HP-PRRSV strain JX143 (pJX143) corresponding to these 88 amino acids. The effect of the deletion on virus replication in cultured cells and transcriptional activation of inflammatory cytokines and chemokines in pulmonary alveolar macrophages (PAMs) was examined. Mutant virus with the 88-aa deletion in nsp2 (rJX143-D88) had faster growth kinetics and produced larger plaques in MARC-145 cells than the parental virus (rJX143), suggesting that the deletion enhanced virus replication in MARC-145 cells. In contrast, the overall yield of rJX143 was almost 1 log higher than that of rJX143-D88, suggesting that the 88-aa deletion in nsp2 decreased the production of infectious viruses in PAMs. Infection with the mutant virus with the 88-aa deletion resulted in increased mRNA expression of type I interferon (IFN-alpha and IFN-beta) and chemokines genes. In addition, the mRNA expression of antiviral genes (ISG15, ISG54 and PKR) regulated by the IFN response was upregulated in PAMs infected with the mutant virus rJX143-D88. Our results demonstrate that virus-specific host immunity can be enhanced by modifying certain nsp2 epitope regions. These findings provide important insights for understanding virus pathogenesis and development of future vaccines. FAU - He, Wei AU - He W AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Wei, Ying AU - Wei Y AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Yao, Jing AU - Yao J AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Xie, Xin AU - Xie X AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Huang, Jiabin AU - Huang J AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Lin, Siyuan AU - Lin S AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Ouyang, Kang AU - Ouyang K AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Chen, Ying AU - Chen Y AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. FAU - Huang, Weijian AU - Huang W AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. weijianghuang@163.com. FAU - Wei, Zuzhang AU - Wei Z AUID- ORCID: 0000-0002-6317-2315 AD - Laboratory of Animal infectious Diseases and Molecular Immunology, College of Animal Science and Technology, Guangxi University, Nanning, 530005, People's Republic of China. zuzhangwei@gxu.edu.cn. LA - eng GR - 31372444/National Natural Science Foundation of China/ GR - 31660716/National Natural Science Foundation of China (CN)/ GR - 2016JJA130049/Guangxi Natural Science Foundation/ GR - XGZ130959/Guangxi University Scientific Research Foundation/ PT - Journal Article DEP - 20180213 PL - Austria TA - Arch Virol JT - Archives of virology JID - 7506870 RN - 0 (Chemokines) RN - 0 (Interferon-alpha) RN - 0 (Transcription Factors) RN - 0 (Ubiquitins) RN - 77238-31-4 (Interferon-beta) RN - EC 2.7.11.1 (eIF-2 Kinase) RN - EC 3.4.22.- (Cysteine Endopeptidases) RN - EC 3.4.22.- (nsP2 proteinase) SB - IM MH - *Amino Acid Sequence MH - Animals MH - Cell Line MH - Cells, Cultured MH - Chemokines/genetics/immunology MH - Chlorocebus aethiops MH - Cysteine Endopeptidases/*genetics/immunology MH - Epithelial Cells/immunology/virology MH - Gene Expression Regulation MH - *Host-Pathogen Interactions MH - Interferon-alpha/genetics/immunology MH - Interferon-beta/genetics/immunology MH - Macrophages, Alveolar/immunology/*virology MH - Porcine respiratory and reproductive syndrome virus/*genetics/growth & development/pathogenicity MH - *Sequence Deletion MH - Signal Transduction MH - Swine MH - Transcription Factors/genetics/immunology MH - Ubiquitins/genetics/immunology MH - Virus Replication/*genetics MH - eIF-2 Kinase/genetics/immunology EDAT- 2018/02/15 06:00 MHDA- 2018/05/31 06:00 CRDT- 2018/02/15 06:00 PHST- 2017/10/25 00:00 [received] PHST- 2018/01/22 00:00 [accepted] PHST- 2018/02/15 06:00 [pubmed] PHST- 2018/05/31 06:00 [medline] PHST- 2018/02/15 06:00 [entrez] AID - 10.1007/s00705-018-3760-7 [pii] AID - 10.1007/s00705-018-3760-7 [doi] PST - ppublish SO - Arch Virol. 2018 Jun;163(6):1489-1501. doi: 10.1007/s00705-018-3760-7. Epub 2018 Feb 13.