Establishment of Systems to Enable Isolation of Porcine Monoclonal Antibodies Broadly Neutralizing the Porcine Reproductive and Respiratory Syndrome Virus

Autor: Jordan E. Young, Jack W.P. Hayes, David Goldeck, Luke P.M. Johnson, Elle L. McLuskey, Katy Moffat, Simon P. Graham, Michael P. Murtaugh, Parimal Roychoudhury, Dana M. Perry, Jean-Pierre Frossard, Raymond R. R. Rowland, Mark J. Kwakkenbos, Arjen Q. Bakker
Jazyk: angličtina
Rok vydání: 2019
Předmět:
0301 basic medicine
lcsh:Immunologic diseases. Allergy
medicine.drug_class
Swine
viruses
animal diseases
Immunology
Porcine Reproductive and Respiratory Syndrome
bcl-X Protein
Heterologous
Monoclonal antibody
Antibodies
Viral
Epitope
Viral vector
Cell Line
03 medical and health sciences
Epitopes
0302 clinical medicine
Antigen
Neutralization Tests
antibody
medicine
heterologous protection
Immunology and Allergy
Animals
Porcine respiratory and reproductive syndrome virus
Neutralizing antibody
Original Research
B-Lymphocytes
B cell
biology
Antibodies
Monoclonal
virus diseases
Viral Vaccines
respiratory system
porcine reproductive and respiratory syndrome virus
Porcine reproductive and respiratory syndrome virus
biology.organism_classification
Virology
Antibodies
Neutralizing
3. Good health
030104 developmental biology
biology.protein
Proto-Oncogene Proteins c-bcl-6
genetic programming
Antibody
lcsh:RC581-607
Immunologic Memory
030215 immunology
Zdroj: Frontiers in Immunology, Vol 10 (2019)
Frontiers in Immunology
ISSN: 1664-3224
Popis: 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.
Databáze: OpenAIRE
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