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. 2016 Mar;10(2):122-6.
doi: 10.1111/irv.12356. Epub 2016 Jan 29.

Impact of a large deletion in the neuraminidase protein identified in a laninamivir-selected influenza A/Brisbane/10/2007 (H3N2) variant on viral fitness in vitro and in ferrets

Julie Ann  1 , Yacine Abed  1 , Edith Beaulieu  2 , Xavier Bouhy  1 , Marie-Hélène Joly  2 , Karen Dubé  2 , Julie Carbonneau  1 , Marie-Eve Hamelin  1 , Corey Mallett  2 , Guy Boivin  1
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Impact of a large deletion in the neuraminidase protein identified in a laninamivir-selected influenza A/Brisbane/10/2007 (H3N2) variant on viral fitness in vitro and in ferrets

Julie Ann et al. Influenza Other Respir Viruses. 2016 Mar.

Abstract

Viral fitness of a laninamivir-selected influenza A/Brisbane/10/2007-like (H3N2) isolate (LRVp9) containing a 237-amino acid neuraminidase deletion and a P194L hemagglutinin mutation was evaluated in vitro and in ferrets. LRVp9 and the wild-type (WT) virus showed comparable replication kinetics in MDCK-ST6GalI cells. Cultured virus was recovered between days 2 and 5 post-infection in nasal washes (NW) from the 4 WT-infected ferrets whereas no virus was recovered from the LRVp9-infected animals. There was a ≥1 log reduction in viral RNA copies/μl of NW for LRVp9 compared to WT at most time points. The large neuraminidase deletion compromises viral infectivity in vivo.

Keywords: A(H3N2); deletion; influenza; neuraminidase; resistance.

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Figures

Figure 1
Figure 1
In vitro replicative capacities of influenza A/Brisbane/10/2007 (H3N2) WT virus and its LRVp9 variant. Viral titers were determined at the indicated time points from supernatants of ST6GalI‐MDCK cells infected at a multiplicity of infection (MOI) of 0·0001. Mean viral titers ± standard deviations from triplicate values of two independent experiments (n = 6) were determined using standard plaque assays.
Figure 2
Figure 2
Nasal wash viral titers of ferrets infected with influenza A/Brisbane/10/2007 (H3N2) WT virus. Viral titers for each WT‐infected ferret were determined using standard plaque assays from nasal wash samples collected at the indicated days post‐inoculation. Note that no infectious virus was recovered from LRVp9‐infected ferrets.
Figure 3
Figure 3
Quantification of viral RNA in nasal wash samples of ferrets infected with influenza A/Brisbane/10/2007 (H3N2) WT virus (A) and its LRVp9 variant (B). The number of viral RNA copies in nasal wash samples collected at the indicated days post‐inoculation was determined by real‐time RT‐PCR targeting the influenza matrix (M) gene.
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