Infection of Differentiated Porcine Airway Epithelial Cells by Influenza Virus: Differential Susceptibility to Infection by Porcine and Avian Viruses

Punyadarsaniya, Darsaniya; Liang, Chi-Jung; Winter, Christine; Petersen, Henning; Rautenschlein, Silke; Hennig-Pauka, Isabel; Schwegmann-Weßels, Christel; Wu, Chung‐Yi; Wong, Chi‐Huey; Herrler, Georg

doi:10.1371/journal.pone.0028429

Cited by 47 publications

(65 citation statements)

References 25 publications

(38 reference statements)

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“…These results correlate with recent reports showing that H9N2 G1 virus replicates efficiently in human cells (117) and that a virus from the G1 lineage can replicate in 4-week-old pigs and in mice (118). In addition, H9N2 viruses from various genotypes can replicate in ferrets (119), and an H9N2 virus grew to a high titer in a porcine differentiated respiratory epithelial cell precision-cut lung slice system (120). Thus, we speculate that H9N2 G1 polymerase is already adapted to mammals.…”

Section: Discussionsupporting

confidence: 91%

Investigation of Influenza Virus Polymerase Activity in Pig Cells

Moncorgé

Long

Cauldwell

et al. 2013

J Virol

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bReassortant influenza viruses with combinations of avian, human, and/or swine genomic segments have been detected frequently in pigs. As a consequence, pigs have been accused of being a "mixing vessel" for influenza viruses. This implies that pig cells support transcription and replication of avian influenza viruses, in contrast to human cells, in which most avian influenza virus polymerases display limited activity. Although influenza virus polymerase activity has been studied in human and avian cells for many years by use of a minigenome assay, similar investigations in pig cells have not been reported. We developed the first minigenome assay for pig cells and compared the activities of polymerases of avian or human influenza virus origin in pig, human, and avian cells. We also investigated in pig cells the consequences of some known mammalian host range determinants that enhance influenza virus polymerase activity in human cells, such as PB2 mutations E627K, D701N, G590S/Q591R, and T271A. The two typical avian influenza virus polymerases used in this study were poorly active in pig cells, similar to what is seen in human cells, and mutations that adapt the avian influenza virus polymerase for human cells also increased activity in pig cells. In contrast, a different pattern was observed in avian cells. Finally, highly pathogenic avian influenza virus H5N1 polymerase activity was tested because this subtype has been reported to replicate only poorly in pigs. H5N1 polymerase was active in swine cells, suggesting that other barriers restrict these viruses from becoming endemic in pigs.

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Section: Discussionsupporting

confidence: 91%

Investigation of Influenza Virus Polymerase Activity in Pig Cells

Moncorgé

Long

Cauldwell

et al. 2013

J Virol

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“…As shown in Fig. 1B, there was no difference in the maximum titer between P1 to P3 viruses, which was about 10 6 PFU/ml, similar to the value reported for the parental virus (32). However, the growth cycle of the P3 virus was shorter than that of the viruses from the previous passages (Fig.…”

supporting

confidence: 81%

“…culture system for differentiated respiratory epithelial cells from the porcine lung that, upon infection by swine influenza viruses (SIV), reflects the viral virulence properties (32,33). To investigate the adaptation of H9N2 avian influenza viruses to growth in the respiratory epithelium of pigs, we passaged an avian influenza virus, A/chicken/Saudi Arabia/CP7/98 (H9N2), three times (P1, P2, and P3) in PLCS.…”

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confidence: 99%

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Mutations during the Adaptation of H9N2 Avian Influenza Virus to the Respiratory Epithelium of Pigs Enhance Sialic Acid Binding Activity and Virulence in Mice

Yang

Punyadarsaniya

Lambertz

et al. 2017

J Virol

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The natural reservoir for influenza viruses is waterfowl, and from there they succeeded in crossing the barrier to different mammalian species. We analyzed the adaptation of avian influenza viruses to a mammalian host by passaging an H9N2 strain three times in differentiated swine airway epithelial cells. Using precisioncut slices from the porcine lung to passage the parental virus, isolates from each of the three passages (P1 to P3) were characterized by assessing growth curves and ciliostatic effects. The only difference noted was an increased growth kinetics of the P3 virus. Sequence analysis revealed four mutations: one each in the PB2 and NS1 proteins and two in the HA protein. The HA mutations, A190V and T212I, were characterized by generating recombinant viruses containing either one or both amino acid exchanges. Whereas the parental virus recognized ␣2,3-linked sialic acids preferentially, the HA190 mutant bound to a broad spectrum of glycans with ␣2,6/8/9-linked sialic acids. The HA212 mutant alone differed only slightly from the parental virus; however, the combination of both mutations (HA190ϩHA212) increased the binding affinity to those glycans recognized by the HA190 mutant. Remarkably, only the HA double mutant showed a significantly increased pathogenicity in mice. In contrast, none of those mutations affected the ciliary activity of the epithelial cells which is characteristic for virulent swine influenza viruses. Taken together, our results indicate that shifts in the HA receptor affinity are just an early adaptation step of avian H9N2 strains; further mutational changes may be required to become virulent for pigs.IMPORTANCE Swine play an important role in the interspecies transmission of influenza viruses. Avian influenza A viruses (IAV) of the H9N2 subtype have successfully infected hosts from different species but have not established a stable lineage. We have analyzed the adaptation of IAV-H9N2 virus to target cells of a new host by passaging the virus three times in differentiated porcine respiratory epithelial cells. Among the four mutations detected, the two HA mutations were analyzed by generating recombinant viruses. Depending on the infection system used, the mutations differed in their phenotypic expression, e.g., sialic acid binding activity, replication kinetics, plaque size, and pathogenicity in inbred mice. However, none of the mutations affected the ciliary activity which serves as a virulence marker. Thus, early

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“…Furthermore, the NS1 protein can increase virus replication by activating the cellular phosphatidylinositol 3-kinase (PI3K) and by downregulation of apoptosis (22,23). The nuclear export protein (NS2/ NEP), which is translated from spliced NS segment mRNA, mediates viral ribonucleoprotein (RNP) export from the nucleus of IV-infected cells via binding to the viral M1 protein (24). Furthermore, the NS2/NEP protein has the ability to modify virus RNA levels by regulation of IV transcription and replication (25).…”

mentioning

confidence: 99%