HA N193D substitution in the HPAI H5N1 virus alters receptor binding affinity and enhances virulence in mammalian hosts.

Seung-Gyu Jang¹ Young-Il Kim² Mark Anthony B Casel^1,3 Jeong Ho Choi¹ Ju Ryeon Gil¹ Rare Rollon¹ Eun-Ha Kim² Se-Mi Kim² Ho Young Ji² Dong Bin Park² Jungwon Hwang⁴ Jae-Woo Ahn² Myung Hee Kim⁴ Min-Suk Song^1,3 Young Ki Choi^1,2,3

Affiliations 4 institutions

College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Republic of Korea.
Center for Study of Emerging and Re-emerging Viruses, Korea Virus Research Institute, Institute for Basic Science (IBS), Daejeon, Republic of Korea.
Zoonotic Infectious Diseases Research Center, Chungbuk National University, Cheongju, Republic of Korea.
Microbiome Convergence Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, Republic of Korea.

PMID38189114 DOI10.1080/22221751.2024.2302854 Year2024 Journal Emerging microbes & infections Emerg Microbes Infect Languageeng Publication statusActive

Active Last checked 2026-10-02 03:30:39 Source: NCBI PubMed

Abstract

During the 2021/2022 winter season, we isolated highly pathogenic avian influenza (HPAI) H5N1 viruses harbouring an amino acid substitution from Asparagine(N) to Aspartic acid (D) at residue 193 of the hemagglutinin (HA) receptor binding domain (RBD) from migratory birds in South Korea. Herein, we investigated the characteristics of the N193D HA-RBD substitution in the A/CommonTeal/Korea/W811/2021[CT/W811] virus by using recombinant viruses engineered via reverse genetics (RG). A receptor affinity assay revealed that the N193D HA-RBD substitution in CT/W811 increases α2,6 sialic acid receptor binding affinity. The rCT/W811-HA_193N virus caused rapid lethality with high virus titres in chickens compared with the rCT/W811-HA_193D virus, while the rCT/W811-HA_193D virus exhibited enhanced virulence in mammalian hosts with multiple tissue tropism. Surprisingly, a ferret-to-ferret transmission assay revealed that rCT/W811-HA_193D virus replicates well in the respiratory tract, at a rate about 10 times higher than that of rCT/W811-HA_193N, and all rCT/W811-HA_193D direct contact ferrets were seroconverted at 10 days post-contact. Further, competition transmission assay of the two viruses revealed that rCT/W811-HA_193D has enhanced growth kinetics compared with the rCT/W811-HA_193N, eventually becoming the dominant strain in nasal turbinates. Further, rCT/W811-HA_193D exhibits high infectivity in primary human bronchial epithelial (HBE) cells, suggesting the potential for human infection. Taken together, the HA-193D containing HPAI H5N1 virus from migratory birds showed enhanced virulence in mammalian hosts, but not in avian hosts, with multi-organ replication and ferret-to-ferret transmission. Thus, this suggests that HA-193D change increases the probability of HPAI H5N1 infection and transmission in humans.

Supporting text Virus Host Location

Clade 2.3.4.4 3 HPAI H5N8 virus 1 Influenza infections 1 pathogenecity 1 reassortant 13 South Korea 5 Influenza A virus 191 Influenza A Virus, H5N1 Subtype 308 Influenza in Birds 352 Animals 1981 Chickens 149 Ferrets 79 Hemagglutinins 24 Humans 1455 Virulence 110

Evidence records

5 total

Zoonotic Surveillance

1 records · 1 evidence types

Evidence type Virus Isolation

1 records

OVE7692 Virus Isolation

Key finding

Highly pathogenic avian influenza H5N1 viruses carrying the HA N193D substitution were isolated from migratory birds in South Korea during the 2021/2022 winter season.

Virus

Host

Location

Supporting text

Experimental Infection

2 records · 2 evidence types

Evidence type Pathogenicity Experiment

1 records

OVE7694 Pathogenicity Experiment

Key finding

The recombinant rCT/W811-HA193N H5N1 virus caused rapid lethality with high virus titres in chickens under controlled infection conditions.

Virus

Host

Location

Supporting text

The rCT/W811-HA193N virus caused rapid lethality with high virus titres in chickens compared with the rCT/W811-HA193D virus.

Method: in vivo infection comparison | viral titre measurement in infected chickens
Experimental system: chicken infection model

Evidence type Experimental Transmission

1 records

OVE7696 Experimental Transmission

Key finding

The rCT/W811-HA193D virus transmitted between ferrets in a direct-contact transmission assay and induced seroconversion in all contact ferrets.

Virus

Host

Location

Supporting text

a ferret-to-ferret transmission assay revealed that rCT/W811-HA193D virus replicates well in the respiratory tract, at a rate about 10 times higher than that of rCT/W811-HA193N, and all rCT/W811-HA193D direct contact ferrets were seroconverted at 10 days post-contact.

Method: ferret-to-ferret transmission assay | direct contact transmission model | seroconversion assessment
Experimental system: ferret-to-ferret direct contact transmission assay

Functional Mechanism

2 records · 2 evidence types

Evidence type Receptor Usage

1 records

OVE7693 Receptor Usage

Key finding

The N193D substitution in the HA receptor binding domain of A/CommonTeal/Korea/W811/2021 (H5N1) increases viral affinity for the α2,6-linked sialic acid receptor.

Virus

Host

Location

Supporting text

A receptor affinity assay revealed that the N193D HA-RBD substitution in CT/W811 increases α2,6 sialic acid receptor binding affinity.

Method: receptor affinity assay
Receptors: α2,6 sialic acid receptor
Host factors: hemagglutinin (HA) receptor binding domain

Evidence type Molecular Adaptation

1 records

OVE7699 Molecular Adaptation

Key finding

The HA N193D substitution in HPAI H5N1 increases α2,6 sialic acid receptor binding affinity and enhances mammalian adaptation and transmission potential.

Virus

Host

Location

Supporting text

A receptor affinity assay revealed that the N193D HA-RBD substitution in CT/W811 increases α2,6 sialic acid receptor binding affinity. ... the rCT/W811-HA193D virus exhibited enhanced virulence in mammalian hosts with multiple tissue tropism. Surprisingly, a ferret-to-ferret transmission assay revealed that rCT/W811-HA193D virus replicates well in the respiratory tract ... Taken together, the HA-193D containing HPAI H5N1 virus from migratory birds showed enhanced virulence in mammalian hosts, but not in avian hosts, with multi-organ replication and ferret-to-ferret transmission.

Genes or proteins: hemagglutinin (HA) | HA receptor binding domain (RBD)
Receptors: α2,6 sialic acid receptor
Host factors: primary human bronchial epithelial (HBE) cells
Mutations: N193D | HA-193D
Mechanism types: receptor binding | host-range expansion | tissue tropism | virulence adaptation | transmission fitness | replication adaptation

References

54 references

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