A Single Amino Acid Substitution in Porcine Reproductive and Respiratory Syndrome Virus Glycoprotein 2 Significantly Impairs Its Infectivity in Macrophages

被引:8
作者
Chaudhari, Jayeshbhai [1 ,2 ]
Leme, Raquel Arruda [1 ,3 ]
Durazo-Martinez, Kassandra [1 ,4 ]
Sillman, Sarah [2 ]
Workman, Aspen M. [5 ]
Vu, Hiep L. X. [1 ,4 ]
机构
[1] Univ Nebraska Lincoln, Nebraska Ctr Virol, Lincoln, NE 68583 USA
[2] Univ Nebraska Lincoln, Sch Vet Med & Biomed Sci, Lincoln, NE 68583 USA
[3] Dechra Pharmaceut, Clinal Res Dept, Londrina, Brazil
[4] Univ Nebraska Lincoln, Dept Anim Sci, Lincoln, NE 68583 USA
[5] United State Dept Agr Agr Res Serv, US Meat Anim Res Ctr, Clay Ctr, NE 68933 USA
来源
VIRUSES-BASEL | 2022年 / 14卷 / 12期
基金
美国食品与农业研究所;
关键词
PRRSV; tropism; PAMs; GP2; CD163; PRRS VIRUS; RECEPTOR; IDENTIFICATION; SIALOADHESIN; INVOLVEMENT; ADAPTATION; VIRULENT; PROTEIN; PIGS;
D O I
10.3390/v14122822
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Porcine reproductive and respiratory syndrome virus (PRRSV) has a restricted tropism for macrophages and CD163 is a key receptor for infection. In this study, the PRRSV strain NCV1 was passaged on MARC-145 cells for 95 passages, and two plaque-clones (C1 and C2) were randomly selected for further analysis. The C1 virus nearly lost the ability to infect porcine alveolar macrophages (PAMs), as well as porcine kidney cells expressing porcine CD163 (PK15-pCD163), while the C2 virus replicates well in these two cell types. Pretreatment of MARC-145 cells with an anti-CD163 antibody nearly blocked C1 virus infection, indicating that the virus still required CD163 to infect cells. The C1 virus carried four unique amino acid substitutions: three in the nonstructural proteins and a K160I in GP2. The introduction of an I160K substitution in GP2 of the C1 virus restored its infectivity in PAMs and PK15-pCD163 cells, while the introduction of a K160I substitution in GP2 of the low-passaged, virulent PRRSV strain NCV13 significantly impaired its infectivity. Importantly, pigs inoculated with the rNCV13-K160I mutant exhibited lower viremia levels and lung lesions than those infected with the parental rNCV13. These results demonstrated that the K160 residue in GP2 is one of the key determinants of PRRSV tropism.
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页数:22
相关论文
共 34 条
[1]   Positively Charged Residues at the Five-Fold Symmetry Axis of Cell Culture-Adapted Foot-and-Mouth Disease Virus Permit Novel Receptor Interactions [J].
Berryman, Stephen ;
Clark, Stuart ;
Kakker, Naresh K. ;
Silk, Rhiannon ;
Seago, Julian ;
Wadsworth, Jemma ;
Chamberlain, Kyle ;
Knowles, Nick J. ;
Jackson, Terry .
JOURNAL OF VIROLOGY, 2013, 87 (15) :8735-8744
[2]   Porcine Alveolar Macrophage-like cells are pro-inflammatory Pulmonary Intravascular Macrophages that produce large titers of Porcine Reproductive and Respiratory Syndrome Virus [J].
Bordet, Elise ;
Maisonnasse, Pauline ;
Renson, Patricia ;
Bouguyon, Edwige ;
Crisci, Elisa ;
Tiret, Mathieu ;
Descamps, Delphyne ;
Bernelin-Cottet, Cindy ;
Urien, Celine ;
Lefevre, Francois ;
Jouneau, Luc ;
Bourry, Olivier ;
Leplat, Jean-Jacques ;
Schwartz-Cornil, Isabelle ;
Bertho, Nicolas .
SCIENTIFIC REPORTS, 2018, 8
[3]   ICTV Virus Taxonomy Profile: Arteriviridae 2021 [J].
Brinton, Margo A. ;
Gulyaeva, Anastasia A. ;
Balasuriya, Udeni B. R. ;
Dunowska, Magda ;
Faaberg, Kay S. ;
Goldberg, Tony ;
Leung, Frederick C. C. ;
Nauwynck, Hans J. ;
Snijder, Eric J. ;
Stadejek, Tomasz ;
Gorbalenya, Alexander E. .
JOURNAL OF GENERAL VIROLOGY, 2021, 102 (08)
[4]  
Burkard C, 2018, J VIROL, V92, DOI [10.1128/JVI.00415-18, 10.1128/jvi.00415-18]
[5]   Precision engineering for PRRSV resistance in pigs: Macrophages from genome edited pigs lacking CD163 SRCR5 domain are fully resistant to both PRRSV genotypes while maintaining biological function [J].
Burkard, Christine ;
Lillico, Simon G. ;
Reid, Elizabeth ;
Jackson, Ben ;
Mileham, Alan J. ;
Ait-Ali, Tahar ;
Whitelaw, C. Bruce A. ;
Archibald, Alan L. .
PLOS PATHOGENS, 2017, 13 (02)
[6]   CD163 expression confers susceptibility to porcine reproductive and respiratory syndrome viruses [J].
Calvert, Jay G. ;
Slade, David E. ;
Shields, Shelly L. ;
Jolie, Rika ;
Mannan, Ramasamy M. ;
Ankenbauer, Robert G. ;
Welch, Siao-Kun W. .
JOURNAL OF VIROLOGY, 2007, 81 (14) :7371-7379
[7]   Identification of Cryptic Promoter Activity in cDNA Sequences Corresponding to PRRSV 5′ Untranslated Region and Transcription Regulatory Sequences [J].
Chaudhari, Jayeshbhai ;
Nguyen, The Nhu ;
Vu, Hiep L. X. .
VIRUSES-BASEL, 2022, 14 (02)
[8]   Porcine Reproductive and Respiratory Syndrome Virus Infection Upregulates Negative Immune Regulators and T-Cell Exhaustion Markers [J].
Chaudhari, Jayeshbhai ;
Liew, Chia-Sin ;
Riethoven, Jean-Jack M. ;
Sillman, Sarah ;
Vu, Hiep L. X. .
JOURNAL OF VIROLOGY, 2021, 95 (21)
[9]   The Minor Envelope Glycoproteins GP2a and GP4 of Porcine Reproductive and Respiratory Syndrome Virus Interact with the Receptor CD163 [J].
Das, Phani B. ;
Dinh, Phat X. ;
Ansari, Israrul H. ;
de Lima, Marcelo ;
Osorio, Fernando A. ;
Pattnaik, Asit K. .
JOURNAL OF VIROLOGY, 2010, 84 (04) :1731-1740
[10]   Involvement of the matrix protein in attachment of porcine reproductive and respiratory syndrome virus to a heparinlike receptor on porcine alveolar macrophages [J].
Delputte, PL ;
Vanderheijden, N ;
Nauwynck, HJ ;
Pensaert, MB .
JOURNAL OF VIROLOGY, 2002, 76 (09) :4312-4320