Virulence of H5N1 virus in mice attenuates after in vitro serial passages

Li, Jing; Liu, Bohua; Chang, Guohui; Hu, Yi; Zhan, Dawei; Xia, Yukun; Li, Yongqiang; Yang, Yinhui; Zhu, Qingqing

doi:10.1186/1743-422x-8-93

Cited by 10 publications

(11 citation statements)

References 36 publications

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“…The mutations of H5 RBD affect not only the viral binding avidity but also the pathogenicity of virus, though the determinants of virulence were usually focused on the NA‐1 and polymerase genes. Our previous studies have indicated the specific mutations in PB1 and PB2 of H5N1 viruses contributed to the upregulated proliferation and attenuation in virulence to mice [Li et al, , ]. In the present study, a recombinant VN1194 virus which carrying mouse lung‐adapted mutation E190G in HA was confirmed to reduce viral replication in vitro and attenuated VN1194 virus in mice.…”

Section: Discussionsupporting

confidence: 68%

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Mouse lung‐adapted mutation of E190G in hemagglutinin from H5N1 influenza virus contributes to attenuation in mice

Han

Sun

et al. 2015

Journal of Medical Virology

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The highly pathogenic H5N1 avian influenza virus is one of the greatest influenza pandemic threats since 2003. The association of the receptor binding domain (RBD) with the virulence of influenza virus is rarely addressed, particularly of H5N1 influenza viruses. In this study, BALB/c mice were intranasally infected with A/Vietnam/1194/2004 (VN1194, H5N1). The mouse lung-adapted variants were isolated and the mutation of E190G (H3 numbering) in the RBD was recognized. The recombinant virus, rVN-E190G carrying E190G in hemagglutinin (HA) was designed and rescued using reverse genetics techniques. The receptor binding activity, growth curve and pathogenicity in mice of the rVN-E190G were investigated. Results demonstrated that rVN-E190G virus increased the binding avidity to α2,6 SA (sialic acid) and reduced the affinity to α2,3 SA, meanwhile weakened the viral replication in vitro. Moreover, the virulence assessment demonstrated that rVN-E190G was attenuated in mice. These results indicated that the mutation E190G in HA decreases H5N1 viral replication in vitro and significantly attenuates virulence in vivo. These findings identify one of the determinants in RBD which can be associated with H5N1 virulence in mice.

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Section: Discussionsupporting

confidence: 68%

“…VN1194 is also highly pathogenic in mice. Mouse model is one of the models of mammalian infection used to assess the pathogenic mechanism and virulence of H5N1 influenza viruses [Li et al, 2011b]. In the current study, the prototype of H5N1 clinical isolate, A/Vietnam/ 1194/2004, had no prior history of mouse passage.…”

Section: Introductionmentioning

confidence: 97%

Mouse lung‐adapted mutation of E190G in hemagglutinin from H5N1 influenza virus contributes to attenuation in mice

Han

Sun

et al. 2015

Journal of Medical Virology

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“…This was previously shown, not only in vitro (2), but also for natural isolates (reviewed in reference 26). Different studies provided hints of the impact of the so-called internal proteins on pathogenicity (16,25). The data presented here additionally imply a possible role of the internal proteins in immunoescape.…”

Section: Discussionmentioning

confidence: 80%

“…One drawback of our system, of course, is the attenuation of the isolates, as reflected in the IVPI. Possibly, this attenuation is caused by alterations in the viral polymerase proteins, as recently speculated (25). In addition, transmissibility as a property of the virus may be reduced without undermining the virus' success in infecting cells, because transmission is guaranteed by the passaging system.…”

Section: Discussionmentioning

confidence: 98%

Highly Pathogenic Avian Influenza Virus Subtype H5N1 Escaping Neutralization: More than HA Variation

Höper

Kalthoff

Hoffmann

2012

J Virol

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Influenza A viruses are one of the major threats in modern health care. Novel viruses arise due to antigenic drift and antigenic shift, leading to escape from the immune system and resulting in a serious problem for disease control. In order to investigate the escape process and to enable predictions of escape, we serially passaged influenza A H5N1 virus in vitro 100 times under immune pressure. The generated escape viruses were characterized phenotypically and in detail by full-genome deep sequencing. Mutations already found in natural isolates were detected, evidencing the in vivo relevance of the in vitro-induced amino acid substitutions. Additionally, several novel alterations were triggered. Altogether, the results imply that our in vitro system is suitable to study influenza A virus evolution and that it might even be possible to predict antigenic changes of influenza A viruses circulating in vaccinated populations.

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“…And our previous study showed that the rescued VN1194, i.e. rVN1194, with C 4 at PB2, PB1 and PA, while U 4 at the other five segments was as high pathogenic as the wild VN1194 virus, with an MLD 50 less than 1 PFU [19], [33]. Because the difference in pathogenicity contributed by the U4/C4 variation in PB1 was not large enough to be identified in the background of rVN1194 (data not shown), we re-rescued the VN1194 virus with C 4 in all segments, which was named as rVN-PB1(C).…”

Section: Discussionmentioning

confidence: 92%

U4 at the 3′ UTR of PB1 Segment of H5N1 Influenza Virus Promotes RNA Polymerase Activity and Contributes to Viral Pathogenicity

Sun

Han

et al. 2014

PLoS ONE

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The viral RNA-dependent RNA polymerase has been found to contribute to efficient replication in mammalian systems and to the high pathogenicity of H5N1 influenza A virus in humans and other mammals. The terminal untranslated regions of the viral segments perform functions such as polyadenylation and contain signals for genomic packaging and initiation of RNA synthesis. These sequences are highly conserved, apart from a U/C polymorphism at position 4 of the 3′ end, most often seen in the polymerase gene segments. However, no study has yet tested whether the untranslated regions of H5N1 make any contribution to its high pathogenicity. Herein, the association of the fourth nucleotide at the 3′ end of the untranslated region in segment 2 (PB1), of A/Vietnam/1194/2004 (H5N1), with pathogenicity was examined in mice. To this end, an RNA polymerase reporter system was constructed, and viruses with mutations at this site were rescued. Results showed the U4 in PB1 was found to contribute to greater amounts of RNA-dependent RNA polymerase activity and differentially regulate genomic transcription and replication. Although a recombinant H5N1 virus with the rarer C4 sequence in all eight segments was viable and replicated to high titers in vitro, replacing a single U4 at the 3′ termini of the PB1 gene segment enhanced viral reproduction and more pathogenesis. In this way, these data showed the importance of untranslated regions of H5N1 influenza virus to pathogenicity.

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Virulence of H5N1 virus in mice attenuates after in vitro serial passages

Cited by 10 publications

References 36 publications

Mouse lung‐adapted mutation of E190G in hemagglutinin from H5N1 influenza virus contributes to attenuation in mice

Mouse lung‐adapted mutation of E190G in hemagglutinin from H5N1 influenza virus contributes to attenuation in mice

Highly Pathogenic Avian Influenza Virus Subtype H5N1 Escaping Neutralization: More than HA Variation

U4 at the 3′ UTR of PB1 Segment of H5N1 Influenza Virus Promotes RNA Polymerase Activity and Contributes to Viral Pathogenicity

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