OmniVira

Cattle and human organoids reveal 2.3.4.4b H5N1 cross-species transmission potential and neuraminidase-specific neutralizing antibodies in humans.

Cun Li1 Yifei Yu1,2 Zhixin Wan1 Jian-Piao Cai1 Jingjing Huang1,2 Xiaoxin Zhu1,2 Jiali Wu1 Wei Xue1 Ying Zhou1,2 Man Chun Chiu1,2 Qiaoshuai Lan1,2 Shuxin Zhang1 Zijun Zhao1 Yidong Yang1 Yuhong Liu1,2 Pui Wang1 Shaofeng Deng1 Ming Yue1 Longchao Zhu3 Keshan Zhang4 Mengmeng Zhao4 Jieshi Yu5 Wenkang Wei5 Xiang Gao6 Lin Huang7 Hin Chu1,2,8,9 Gang Wu10 Honglin Chen1,2,8,9 Kwok Yung Yuen11,12,13,14,15,16,17 Richard Webby18 Jie Zhou19,20,21,22,23
Affiliations 23 institutions
  1. Department of Microbiology, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China.
  2. Centre for Virology, Vaccinology and Therapeutics, Hong Kong Science and Technology Park, Hong Kong Special Administrative Region, China.
  3. School of Life and Health Technology, Dongguan University of Technology, Dongguan, China.
  4. Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, School of Animal Science and Technology, Foshan University, Foshan, China.
  5. State Key Laboratory of Swine and Poultry Breeding Industry, Agro-Biological Gene Research Center, Guangdong Academy of Agricultural Sciences, Guangzhou, China.
  6. Laboratory of Advanced Biotechnology, Beijing Institute of Biotechnology, Beijing, China.
  7. BiomOrgan Ltd, Hong Kong, China.
  8. State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China.
  9. Pandemic Research Alliance Unit at The University of Hong Kong, Hong Kong Special Administrative Region, China.
  10. Department of Host Microbe Interactions, St. Jude Children's Research Hospital, Memphis, TN, USA.
  11. Department of Microbiology, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
  12. Centre for Virology, Vaccinology and Therapeutics, Hong Kong Science and Technology Park, Hong Kong Special Administrative Region, China. [email protected].
  13. State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
  14. Pandemic Research Alliance Unit at The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
  15. Carol Yu Centre for Infection, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
  16. Department of Infectious Disease and Microbiology, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China. [email protected].
  17. Department of Microbiology, Queen Mary Hospital, Pokfulam, Hong Kong Special Administrative Region, China. [email protected].
  18. Department of Host Microbe Interactions, St. Jude Children's Research Hospital, Memphis, TN, USA. [email protected].
  19. Department of Microbiology, School of Clinical Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
  20. Centre for Virology, Vaccinology and Therapeutics, Hong Kong Science and Technology Park, Hong Kong Special Administrative Region, China. [email protected].
  21. BiomOrgan Ltd, Hong Kong, China. [email protected].
  22. State Key Laboratory of Emerging Infectious Diseases, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
  23. Pandemic Research Alliance Unit at The University of Hong Kong, Hong Kong Special Administrative Region, China. [email protected].
PMID42420291 DOI10.1038/s41467-026-74345-w Year2026 Journal Nature communications Nat Commun Languageeng Statusepublish

Abstract

The unexpected circulation of clade 2.3.4.4b H5N1 influenza viruses in dairy cattle and the transmission to diverse mammalian species poses a pandemic risk. We sought to explore cattle and human respiratory susceptibility to the 2.3.4.4b H5N1 virus. We establish long-term expandable cattle airway and mammary organoids. The 2.3.4.4b H5N1 virus exhibits high replicative fitness in cattle mammary organoids, recapitulating its remarkable mammary tropism. The virus also replicates robustly in cattle airway organoids, suggesting an underrecognized respiratory component in ongoing outbreaks. Interestingly, human airway and nasal organoids are highly susceptible to the 2.3.4.4b H5N1 virus. Yet, a novel organoid-based neutralization assay reveals that N1 antibodies in human sera had cross-neutralizing activity against the 2.3.4.4b H5N1 and ancestral H5N1-VN1194 viruses. The cross-neutralization, exclusively manifested in the organoid-based assay, is enhanced after seasonal influenza vaccination and diminished after depleting N1-specific antibodies. Therefore, cross-neutralizing N1 antibodies are likely limiting zoonotic infection by H5N1 viruses in humans.

Supporting text Virus Host Location
Antibodies, Neutralizing 80 Antibodies, Viral 212 Influenza A Virus, H5N1 Subtype 300 Influenza, Human 286 Neuraminidase 62 Organoids 4 Orthomyxoviridae Infections 228 Animals 1948 Cattle 126 Cross Reactions 21 Female 289 Humans 1440 Neutralization Tests 31

Evidence records

6 total
Zoonotic Surveillance
3 records · 1 evidence types
Evidence type Serological Evidence
3 records
OVE11742 Serological Evidence
Key finding

Human sera contain N1 antibodies that cross-neutralize clade 2.3.4.4b H5N1 and ancestral H5N1-VN1194 in an organoid-based neutralization assay.

Virus
Influenza A virus H5N1 H5N1-VN1194
Host
human Homo sapiens Natural host
Location
Not specified
Supporting text

a novel organoid-based neutralization assay reveals that N1 antibodies in human sera had cross-neutralizing activity against the 2.3.4.4b H5N1 and ancestral H5N1-VN1194 viruses.

Method
organoid-based neutralization assay
Sample type
sera
OVE11743 Serological Evidence
Key finding

Cross-neutralizing activity in human sera is enhanced after seasonal influenza vaccination in the organoid-based assay.

Virus
Influenza A virus H5N1 H5N1-VN1194
Host
human Homo sapiens Natural host
Location
Not specified
Supporting text

The cross-neutralization, exclusively manifested in the organoid-based assay, is enhanced after seasonal influenza vaccination

Method
organoid-based neutralization assay
Sample type
sera
OVE11744 Serological Evidence
Key finding

Depletion of N1-specific antibodies from human sera diminishes cross-neutralization against H5N1 in the organoid-based assay.

Virus
Influenza A virus H5N1 H5N1-VN1194
Host
human Homo sapiens Natural host
Location
Not specified
Supporting text

and diminished after depleting N1-specific antibodies.

Method
organoid-based neutralization assay | antibody depletion
Sample type
sera
Experimental Infection
3 records · 1 evidence types
Evidence type Host Range Experiment
3 records
OVE11739 Host Range Experiment
Key finding

Clade 2.3.4.4b H5N1 replicates with high fitness in cattle mammary organoids, indicating strong mammary tropism.

Virus
Influenza A virus H5N1
Host
cattle mammary organoids Experimental system
Location
Not specified
Supporting text

The 2.3.4.4b H5N1 virus exhibits high replicative fitness in cattle mammary organoids, recapitulating its remarkable mammary tropism.

Method
organoid infection assay | viral replication assessment
Experimental system
organoid culture (cattle mammary organoids)
OVE11740 Host Range Experiment
Key finding

Clade 2.3.4.4b H5N1 replicates robustly in cattle airway organoids.

Virus
Influenza A virus H5N1
Host
cattle airway organoids Experimental system
Location
Not specified
Supporting text

The virus also replicates robustly in cattle airway organoids, suggesting an underrecognized respiratory component in ongoing outbreaks.

Method
organoid infection assay | viral replication assessment
Experimental system
organoid culture (cattle airway organoids)
OVE11741 Host Range Experiment
Key finding

Human airway and nasal organoids are highly susceptible to clade 2.3.4.4b H5N1.

Virus
Influenza A virus H5N1
Host
human Homo sapiens Experimental system
Location
Not specified
Supporting text

Interestingly, human airway and nasal organoids are highly susceptible to the 2.3.4.4b H5N1 virus.

Method
organoid infection assay | viral susceptibility/replication assessment
Experimental system
organoid culture (human airway and nasal organoids)