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Establishment of systems to enable isolation of porcine monoclonal antibodies broadly neutralizing the porcine reproductive and respiratory syndrome virus

Goldeck, David, Perry, Dana, Hayes, Jack, Johnson, Luke, Young, Jordan, Roychoudhury, Parimal, McLuskey, Elle, Moffat, Katy, Bakker, Arjen, Kwakkenbos, Mark , Frossard, Jean-Pierre, Rowland, Bob, Murtaugh, Michael and Graham, Simon P. (2019) Establishment of systems to enable isolation of porcine monoclonal antibodies broadly neutralizing the porcine reproductive and respiratory syndrome virus Frontiers in Immunology.

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Abstract

The rapid evolution of porcine reproductive and respiratory syndrome viruses (PRRSV) poses a major challenge to effective disease control since available vaccines show variable efficacy against divergent strains. Knowledge of the antigenic targets of virus-neutralizing antibodies that confer protection against heterologous PRRSV strains would be a catalyst for the development of next-generation vaccines. Key to discovering these epitopes is the isolation of neutralizing monoclonal antibodies (mAbs) from immune pigs. To address this need, we sought to establish systems to enable the isolation of PRRSV neutralizing porcine mAbs. We experimentally produced a cohort of immune pigs by sequential challenge infection with four heterologous PRRSV strains spanning PRRSV-1 subtypes and PRRSV species. Whilst priming with PRRSV-1 subtype 1 did not confer full protection against a subsequent infection with a PRRSV-1 subtype 3 strain, animals were protected against a subsequent PRRSV-2 infection. The infection protocol resulted in high serum neutralizing antibody titers against PRRSV-1 Olot/91 and significant neutralization of heterologous PRRSV-1/-2 strains. Enriched memory B cells isolated at the termination of the study were genetically programmed by transduction with a retroviral vector expressing the Bcl-6 transcription factor and the anti-apoptotic Bcl-xL protein, a technology we demonstrated efficiently converts porcine memory B cells into proliferating antibody-secreting cells. Pools of transduced memory B cells were cultured and supernatants containing PRRSV-specific antibodies identified by flow cytometric staining of infected MARC-145 cells and in vitro neutralization of PRRSV-1. Collectively, these data suggest that this experimental system may be further exploited to produce a panel of PRRSV-specific mAbs, which will contribute both to our understanding of the antibody response to PRRSV and allow epitopes to be resolved that may ultimately guide the design of immunogens to induce cross-protective immunity.

Item Type: Article
Divisions : Faculty of Health and Medical Sciences > School of Veterinary Medicine
Authors :
NameEmailORCID
Goldeck, David
Perry, Danad.perry@surrey.ac.uk
Hayes, Jack
Johnson, Lukel.p.johnson@surrey.ac.uk
Young, Jordan
Roychoudhury, Parimal
McLuskey, Elle
Moffat, Katy
Bakker, Arjen
Kwakkenbos, Mark
Frossard, Jean-Pierre
Rowland, Bob
Murtaugh, Michael
Graham, Simon P.s.graham@surrey.ac.uk
Date : 2019
DOI : 10.3389/fimmu.2019.00572
Copyright Disclaimer : © 2019 Goldeck, Perry, Hayes, Johnson, Young, Roychoudhury, McLuskey, Moffat, Bakker, Kwakkenbos, Frossard, Rowland, Murtaugh and Graham. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Uncontrolled Keywords : Porcine Reproductive and Respiratory Syndrome Virus; B cell; Antibody; Heterologous protection; Genetic programming
Depositing User : Clive Harris
Date Deposited : 21 Mar 2019 10:09
Last Modified : 21 Mar 2019 10:09
URI: http://epubs.surrey.ac.uk/id/eprint/850827

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