A Triple Amino Acid Substitution at Position 88/94/95 in Glycoprotein GP2a of Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV1) Is Responsible for Adaptation to MARC-145 Cells

Jiexiong Xie, Ivan Trus, Dayoung Oh, Lise Kirstine Kvisgaard, Julie C F Rappe, Nicolas Ruggli, Nathalie Vanderheijden, Lars Erik Larsen, François Lefèvre, Hans J Nauwynck*

*Corresponding author for this work

    Research output: Contribution to journalJournal articleResearchpeer-review

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    Abstract

    The Meat Animal Research Center-145 (MARC-145) cell line has been proven to be valuable for viral attenuation regarding vaccine development and production. Cell-adaptation is necessary for the efficient replication of porcine reproductive and respiratory syndrome virus (PRRSV) in these cells. Multiple sequence analysis revealed consistent amino acid substitutions in GP2a (V88F, M94I, F95L) of MARC-145 cell-adapted strains. To investigate the putative effect of these substitutions, mutations at either position 88, 94, 95, and their combinations were introduced into two PRRSV1 (13V091 and IVI-1173) infectious clones followed by the recovery of viable recombinants. When comparing the replication kinetics in MARC-145 cells, a strongly positive effect on the growth characteristics of the 13V091 strain (+2.1 log10) and the IVI-1173 strain (+1.7 log10) compared to wild-type (WT) virus was only observed upon triple amino acid substitution at positions 88 (V88F), 94 (M94I), and 95 (F95L) of GP2a, suggesting that the triple mutation is a determining factor in PRRSV1 adaptation to MARC-145 cells.
    Original languageEnglish
    Article number36
    JournalViruses
    Volume11
    Issue number1
    Number of pages17
    ISSN1999-4915
    DOIs
    Publication statusPublished - 2019

    Keywords

    • GP2a
    • MARC-145
    • PRRSV
    • adaptation mutation

    Cite this

    Xie, Jiexiong ; Trus, Ivan ; Oh, Dayoung ; Kvisgaard, Lise Kirstine ; Rappe, Julie C F ; Ruggli, Nicolas ; Vanderheijden, Nathalie ; Larsen, Lars Erik ; Lefèvre, François ; Nauwynck, Hans J. / A Triple Amino Acid Substitution at Position 88/94/95 in Glycoprotein GP2a of Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV1) Is Responsible for Adaptation to MARC-145 Cells. In: Viruses. 2019 ; Vol. 11, No. 1.
    @article{9eb384223a9049819c15f9d056e8477f,
    title = "A Triple Amino Acid Substitution at Position 88/94/95 in Glycoprotein GP2a of Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV1) Is Responsible for Adaptation to MARC-145 Cells",
    abstract = "The Meat Animal Research Center-145 (MARC-145) cell line has been proven to be valuable for viral attenuation regarding vaccine development and production. Cell-adaptation is necessary for the efficient replication of porcine reproductive and respiratory syndrome virus (PRRSV) in these cells. Multiple sequence analysis revealed consistent amino acid substitutions in GP2a (V88F, M94I, F95L) of MARC-145 cell-adapted strains. To investigate the putative effect of these substitutions, mutations at either position 88, 94, 95, and their combinations were introduced into two PRRSV1 (13V091 and IVI-1173) infectious clones followed by the recovery of viable recombinants. When comparing the replication kinetics in MARC-145 cells, a strongly positive effect on the growth characteristics of the 13V091 strain (+2.1 log10) and the IVI-1173 strain (+1.7 log10) compared to wild-type (WT) virus was only observed upon triple amino acid substitution at positions 88 (V88F), 94 (M94I), and 95 (F95L) of GP2a, suggesting that the triple mutation is a determining factor in PRRSV1 adaptation to MARC-145 cells.",
    keywords = "GP2a, MARC-145, PRRSV, adaptation mutation",
    author = "Jiexiong Xie and Ivan Trus and Dayoung Oh and Kvisgaard, {Lise Kirstine} and Rappe, {Julie C F} and Nicolas Ruggli and Nathalie Vanderheijden and Larsen, {Lars Erik} and Fran{\cc}ois Lef{\`e}vre and Nauwynck, {Hans J}",
    year = "2019",
    doi = "10.3390/v11010036",
    language = "English",
    volume = "11",
    journal = "Viruses",
    issn = "1999-4915",
    publisher = "M D P I AG",
    number = "1",

    }

    A Triple Amino Acid Substitution at Position 88/94/95 in Glycoprotein GP2a of Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV1) Is Responsible for Adaptation to MARC-145 Cells. / Xie, Jiexiong; Trus, Ivan; Oh, Dayoung; Kvisgaard, Lise Kirstine; Rappe, Julie C F; Ruggli, Nicolas; Vanderheijden, Nathalie; Larsen, Lars Erik; Lefèvre, François; Nauwynck, Hans J.

    In: Viruses, Vol. 11, No. 1, 36, 2019.

    Research output: Contribution to journalJournal articleResearchpeer-review

    TY - JOUR

    T1 - A Triple Amino Acid Substitution at Position 88/94/95 in Glycoprotein GP2a of Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV1) Is Responsible for Adaptation to MARC-145 Cells

    AU - Xie, Jiexiong

    AU - Trus, Ivan

    AU - Oh, Dayoung

    AU - Kvisgaard, Lise Kirstine

    AU - Rappe, Julie C F

    AU - Ruggli, Nicolas

    AU - Vanderheijden, Nathalie

    AU - Larsen, Lars Erik

    AU - Lefèvre, François

    AU - Nauwynck, Hans J

    PY - 2019

    Y1 - 2019

    N2 - The Meat Animal Research Center-145 (MARC-145) cell line has been proven to be valuable for viral attenuation regarding vaccine development and production. Cell-adaptation is necessary for the efficient replication of porcine reproductive and respiratory syndrome virus (PRRSV) in these cells. Multiple sequence analysis revealed consistent amino acid substitutions in GP2a (V88F, M94I, F95L) of MARC-145 cell-adapted strains. To investigate the putative effect of these substitutions, mutations at either position 88, 94, 95, and their combinations were introduced into two PRRSV1 (13V091 and IVI-1173) infectious clones followed by the recovery of viable recombinants. When comparing the replication kinetics in MARC-145 cells, a strongly positive effect on the growth characteristics of the 13V091 strain (+2.1 log10) and the IVI-1173 strain (+1.7 log10) compared to wild-type (WT) virus was only observed upon triple amino acid substitution at positions 88 (V88F), 94 (M94I), and 95 (F95L) of GP2a, suggesting that the triple mutation is a determining factor in PRRSV1 adaptation to MARC-145 cells.

    AB - The Meat Animal Research Center-145 (MARC-145) cell line has been proven to be valuable for viral attenuation regarding vaccine development and production. Cell-adaptation is necessary for the efficient replication of porcine reproductive and respiratory syndrome virus (PRRSV) in these cells. Multiple sequence analysis revealed consistent amino acid substitutions in GP2a (V88F, M94I, F95L) of MARC-145 cell-adapted strains. To investigate the putative effect of these substitutions, mutations at either position 88, 94, 95, and their combinations were introduced into two PRRSV1 (13V091 and IVI-1173) infectious clones followed by the recovery of viable recombinants. When comparing the replication kinetics in MARC-145 cells, a strongly positive effect on the growth characteristics of the 13V091 strain (+2.1 log10) and the IVI-1173 strain (+1.7 log10) compared to wild-type (WT) virus was only observed upon triple amino acid substitution at positions 88 (V88F), 94 (M94I), and 95 (F95L) of GP2a, suggesting that the triple mutation is a determining factor in PRRSV1 adaptation to MARC-145 cells.

    KW - GP2a

    KW - MARC-145

    KW - PRRSV

    KW - adaptation mutation

    U2 - 10.3390/v11010036

    DO - 10.3390/v11010036

    M3 - Journal article

    VL - 11

    JO - Viruses

    JF - Viruses

    SN - 1999-4915

    IS - 1

    M1 - 36

    ER -