J Bacteriol Virol.  2012 Dec;42(4):330-338. 10.4167/jbv.2012.42.4.330.

Generation and Biological Characterization of a Neutralization-Resistant Mutant of Newcastle Disease Virus

• Park MJ
• Kye SJ
• Kim JY
• Kim S
• Seul HJ
• Park CK
• Choi KS

• KMID: 2168668
• DOI: http://doi.org/10.4167/jbv.2012.42.4.330

Abstract

A neutralization-resistant mutant of Newcastle disease virus (NDV) Kr005 strain belonging to class II genotype VII was generated using a neutralizing monoclonal antibody and its biological effects were assessed. The mutant showed single amino acid substitution (E to K) at position 347 of the hemagglutinin-neuraminidase (HN) protein (E347K mutant). The E347K mutant exhibited marked rounding of the cells and few syncytia in infected chicken embryofibroblast (CEF) cells. The hemadsorption and neuraminidase activities of the E347K mutant of the wild-type virus were 118% and 166%, respectively. The mutant produced a rapid elution pattern whereas the wild type had a slow elution pattern. Growth kinetics studies showed that the E347K mutant produced an 80-times higher yield of extracellular virus in CEF cells compared with the wild-type virus. The time-course virus titer showed a marked increase in mutant-infected cells from 6 h to 12 h post infection (pi), which was consistent with the titer pattern time-course for NA activity. The E347K mutant virus showed a slight decrease in virulence compared to the wild-type virus, but there was no change in pathotype when measured by in vivo pathogenicity testing. These results suggest that an E347K mutation in HN protein might be associated with increased NA activity and subsequent enhancement of virus release from infected cells without change in viral pathotype.

Keyword

Newcastle disease virus; HN protein; Neuraminidase activity; Neutralization resistant escape mutant

MeSH Terms

Amino Acid Substitution
Animals
Chickens
Genotype
Giant Cells
Hemadsorption
HN Protein
Kinetics
Neuraminidase
Newcastle Disease
Newcastle disease virus
Sprains and Strains
Viral Load
Virus Release
Viruses
HN Protein
Neuraminidase

Figure

• 

  Figure 1 Cytopathic effects induced by wild-type and E347K mutant viruses at 48 dpi in CEF cells. (A) wild type Kr005-infected cells; (B) E347K mutant-infected cells; (C) mock-infected control. E347K mutant virus was generated after 10 passages of wild-type NDV Kr005 strain in the primary CEF cells in the presence of mAb 10F11. Arrow represents cytopathic effects. E347K mutant-infected cells were rounded and granulated with few small-sized few syncytia while the wild type Kr005- infected cells developed large-sized syncytia in primary CEF cells.

• 

  Figure 2 Growth curve kinetics (A) and time-course neuraminidase activity (B) of extracellular viruses released from CEF cells infected with E347K mutant or wild-type viruses. Wild-type and E347K mutant viruses were generated after 10 passages of NDV Kr005 strain in the primary CEF cells in the absence or presence of mAb 10F11, respectively. The E347K mutant virus showed higher growth and enhanced NA activity in primary CEF cells compared to the wild-type virus.

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