Thursday, December 09, 2021

PLoS Path: H9N2 Virus-derived M1 Protein Promotes H5N6 Virus Release in Mammalian Cells


For a ubiquitous, low pathogenic, and relatively innocuous avian flu virus - one that rarely infects, and usually only mildly sickens humans - we spend an awful lot of time looking at, and trying to analyze, LPAI H9N2.

On its own, H9N2 has shown only limited pandemic potential, and is not expected to have the same impact as an H5 or H7 avian flu. Still, in May of 2020 the CDC added a new (H9N2 Y280 lineage [A/Anhui-Lujiang/13/2018]) to their short list of zoonotic novel flu viruses they are monitoring. 

Where H9N2 shines is in its ability to reassort with other novel flu viruses, frequently lending its internal genes to some of the most dangerous avian flu viruses (H5N1, H7N9, H10N8) on the planet. A few (of dozens) of past studies include:

H9N2's promiscuity has helped to create a diverse and growing array of H5 and H7 viruses, some (even of the same subtype) being more pathogenic in humans than others (see Differences In Virulence Between Closely Related H5N1 Strains).

H9N2 continues to spread globally, having expanded its range to both the Middle East and Africa over the past decade.  

This spread, and continual evolution of H9N2 - often aided and abetted by the use of older, less effective poultry vaccines (see J. Virus Erad.: Ineffective Control Of LPAI H9N2 By Inactivated Poultry Vaccines - China) - provides more opportunities for H9N2 to encounter, and reassort with, new influenza viruses. 

The Asian avian H5N6 virus is a novel avian flu virus with a far more virulent track record in humans, although it has yet to develop the ability to transmit efficiently from human-to-human. So far there are only 53 known human infections, 52 of which have occurred in Mainland China. 

Not all avian H5N6 viruses are able to infect, or sicken, humans (see Nature Sci Rpts: H5N6 Viruses Exhibit Varying Pathogenicity & Transmissibility In Mammals)But several subclades can, and the virus continues to evolve, primarily through reassortment. 

Last September the CCDC Weekly published a new study looking at two human H5N6 infections this year in Guangxi Zhuang Autonomous Region, which occurred 6 months apart (Feb. & July).  

Both viruses were of different subclades ( &, were the product of recent (and different) ressortments, and showed worrisome mutations which could increase their threat. 

Given the recent increased level of H5N6 activity in China, this avian flu virus is suddenly back on everyone's radar (see CDC Adds A New H5N6 Avian Flu Virus To IRAT List). 

Today we have a new study, published in PloS Pathogens, that looks at the apparent impact of H9N2's matrix protein 1 (M1) gene on H5N6's ability to infect mammals, including humans. 

I've only posted the Abstract, and a few excerpts, from a much longer, and more technical discussion below.  Follow the link to read it in its entirety. 

H9N2 virus-derived M1 protein promotes H5N6 virus release in mammalian cells: Mechanism of avian influenza virus inter-species infection in humans

Fangtao Li, Jiyu Liu, Jizhe Yang, Haoran Sun, Zhimin Jiang, Chenxi Wang, Xin Zhang, Yinghui Yu, Chuankuo Zhao, Juan Pu, Yipeng Sun, Kin-Chow Chang, Jinhua Liu , Honglei Sun

Published: December 3, 2021


H5N6 highly pathogenic avian influenza virus (HPAIV) clade not only exhibits unprecedented intercontinental spread in poultry, but can also cause serious infection in humans, posing a public health threat.
Phylogenetic analyses show that 40% (8/20) of H5N6 viruses that infected humans carried H9N2 virus-derived internal genes. However, the precise contribution of H9N2 virus-derived internal genes to H5N6 virus infection in humans is unclear.
Here, we report on the functional contribution of the H9N2 virus-derived matrix protein 1 (M1) to enhanced H5N6 virus replication capacity in mammalian cells. Unlike H5N1 virus-derived M1 protein, H9N2 virus-derived M1 protein showed high binding affinity for H5N6 hemagglutinin (HA) protein and increased viral progeny particle release in different mammalian cell lines. Human host factor, G protein subunit beta 1 (GNB1), exhibited strong binding to H9N2 virus-derived M1 protein to facilitate M1 transport to budding sites at the cell membrane. GNB1 knockdown inhibited the interaction between H9N2 virus-derived M1 and HA protein, and reduced influenza virus-like particles (VLPs) release.
Our findings indicate that H9N2 virus-derived M1 protein promotes avian H5N6 influenza virus release from mammalian, in particular human cells, which could be a major viral factor for H5N6 virus cross-species infection.

Author summary

H9N2 avian influenza viruses (AIVs), through reassortment of their internal genes with other circulating AIVs subtypes (H5N6, H7N9, H10N8 and H10N3 viruses), are known to enable the resulting novel reassortants to infect humans. However, the precise roles of H9N2 virus-derived internal genes in human infection remain unknown.
Here, we found that H9N2 virus-derived (but not H5N1 virus-derived) M1 protein showed strong interaction with host GNB1 protein to enhance M1 protein transport to budding sites at the cell membrane to promote progeny virus release, hence facilitating H5N6 reassortants to infect human cells.



H9N2 AIVs are a major threat to human public health by acting as donors of viral genes through reassortment with co-circulating influenza viruses. Here, we demonstrated that H9N2 virus-derived M1, but not H5N1 virus-derived M1, enhances interaction with human host factor GNB1, to promote M1 transport to the cell membrane which, in turn, facilitates the interaction between M1 and HA proteins, ultimately improving viral assembly and release. Our results suggest that H9N2 virus-derived M1 protein, along with surface HA, is key to increased incidence of H5N6 influenza virus infection in humans.

          (Continue . . . )

While it may never happen - or it may take years before HPAI H5N6 evolves enough to pose a legitimate public health threat - another pandemic is considered inevitable (see PNAS Research: Intensity and Frequency of Extreme Novel Epidemics), and it could easily be worse than COVID.

Which is why we should be preparing seriously now, lest we risk being caught flat footed and unprepared.