Prior #immunity to seasonal #influenza #H3N2 virus confers varying levels of cross - #protection against challenge with clade 2.3.4.4b #H5N1, #H7N9, or #H9N2 virus in a #ferret model

 

ABSTRACT

Evaluating how prior immunity to seasonal influenza viruses influences subsequent zoonotic influenza A virus (IAV) infection in animal models is critical for pandemic preparedness. In this study, we investigated the cross-protective effect of pre-existing A(H3N2) immunity in ferrets challenged with three distinct subtypes of zoonotic IAVs: low pathogenic A(H7N9) and A(H9N2) viruses, and highly pathogenic clade 2.3.4.4b A(H5N1) virus. Our results show that A(H3N2) preimmunity conferred some protection against A(H5N1) and A(H9N2) virus infection, as evidenced by more rapid viral clearance in the upper respiratory tract, reduced virus shedding in the nasal wash on select days post-inoculation, and a lowered frequency of viral detection in specific tissues compared with naive animals. In contrast, A(H3N2) preimmunity provided minimal cross-protection against A(H7N9) infection, as weight loss and viral dissemination in tissues were not significantly reduced in A(H3N2) preimmune ferrets relative to naive animals. These findings highlight the variable breadth and magnitude of cross-protection elicited by prior seasonal IAV immunity against zoonotic influenza virus challenges in the ferret model. Seasonal influenza A(H3N2) preimmunity provided differing levels of cross-protection against zoonotic influenza A virus infections in ferrets.

Source: 

Link: https://journals.asm.org/doi/10.1128/spectrum.03974-25

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Evaluation of Cross-Immunogenicity of #Ferret #Antisera Following Immunization with #H5N1 #Vaccine Strains

 

Abstract

Background: 

Highly pathogenic avian influenza H5N1 viruses of clade 2.3.4.4b have spread globally since 2021, causing extensive outbreaks in avian populations and repeated spillovers into diverse mammalian hosts, including humans. These cross-species transmission events highlight ongoing pandemic risks and underscore the need for vaccine strategies that reflect viral evolution at the human–animal interface. Despite the availability of licensed H5 vaccines and newly recommended World Health Organization (WHO) candidate vaccine viruses (CVVs), the extent to which these vaccines elicit cross-reactive antibody responses against contemporary clade 2.3.4.4b viruses, including mammalian spillover isolates of avian origin, remains incompletely characterized. 

Method: 

In this study, ferret antisera were generated using four WHO-recommended H5 CVVs, including a clade 1 strain (A/Vietnam/1194/2004) and three clade 2.3.4.4b strains (A/Astrakhan/3212/2020, A/American wigeon/South Carolina/22-000345-001/2021, and A/Ezo red fox/Hokkaido/1/2022), formulated with alum adjuvant to reflect licensed vaccine formulation used in national preparedness programs. Antibody responses and cross-reactive activity were evaluated using hemagglutination inhibition (HI) and microneutralization (MN) assays against homologous vaccine strains and a feline-origin clade 2.3.4.4b H5N1 field isolate from Korea, A/Feline/Korea/SNU-01/2023. 

Results: 

Antisera induced by clade 2.3.4.4b CVVs showed cross-reactive antibody responses against homologous and heterologous clade 2.3.4.4b viruses and demonstrated measurable HI and MN responses against the feline-origin field isolate. In contrast, antisera raised against the clade 1 Vietnam CVV exhibited limited cross-reactivity against clade 2.3.4.4b viruses. Overall, clade 2.3.4.4b CVVs generally showed higher antibody responses than the clade 1 vaccine strain across multiple panels. 

Conclusions: 

These findings provide descriptive insights into antigenic differences between clade 1 and clade 2.3.4.4b viruses and support the antigenic relevance of clade 2.3.4.4b CVVs for contemporary H5N1 strains. This study highlights the importance of ongoing antigenic evaluation to inform vaccine strain selection within a One Health framework.

Source: 

Link: https://www.mdpi.com/2076-393X/14/4/301

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Cross - #protection against highly pathogenic avian #influenza #H5N1 virus from seasonal influenza #vaccines: a systematic review and meta-analysis of #ferret studies

 

ABSTRACT

The recent surge in spillover events of highly pathogenic avian influenza A(H5N1) clade 2.3.4.4b to humans and mammals in North America has raised urgent pandemic concerns. Human H5N1 vaccines are unavailable in most countries. We synthesized data from ferret challenge trials to evaluate whether widely available seasonal influenza vaccines confer cross-protection against lethal H5N1 infection. We systematically searched PubMed, Embase, and Web of Science for ferret studies of lethal H5N1 challenge published up to 5 July 2025 (PROSPERO #CRD42024520346). Random-effects meta-analyses were conducted to compare vaccine efficacy (VE) of seasonal influenza vaccines and H5N1 vaccines against H5N1-related mortality. Seroprotection was defined as a neutralizing antibody titre of ≥1:40. We identified 35 studies (157 trials). Seasonal influenza vaccines without N1 did not confer significant cross-protection (five trials; VE 14.8%, 95% CI –3.6 to 30.0). In contrast, VE was 73% for N1-containing seasonal influenza vaccines (19 trials; 95% CI 54–84) and 77% for H5N1 vaccines overall (133 trials; 95% CI 72–82) (p = 0.52). The VE of N1-containing seasonal influenza vaccines was modestly lower than that of H5N1 vaccines with seroprotection (88%; 66 trials; 95% CI 84–91; p = 0.009), but comparable to H5N1 vaccines that did not achieve seroprotection (63%; 67 trials; 95% CI 52–71; p = 0.29). The VE of seasonal influenza vaccines against H5N1 was robust across sensitivity analyses, with no evidence of publication bias (p = 0.99). Seasonal influenza vaccines significantly reduce H5N1-associated mortality in ferret trials, suggesting the cross-protection potential of currently available vaccines. Human studies are warranted.

Source: 

Link: https://www.tandfonline.com/doi/full/10.1080/22221751.2026.2654278

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MF59-adjuvanted A/Astrakhan #influenza #vaccine induces cross-neutralizing #H5N1 #antibodies in #ferrets against circulating clade 2.3.4.4b viruses

 

Abstract

The continued global spread of highly pathogenic avian influenza A(H5N1) viruses, particularly clade 2.3.4.4b, has increased zoonotic spillover risk and underscored the urgency of pandemic preparedness. Human vaccination is a key strategy for mitigating severe disease and limiting transmission, especially in a setting where avian influenza viruses pose a zoonotic threat. We evaluated the immunogenicity of the MF59-adjuvanted, egg-derived A/Astrakhan/3212/2020 (H5N8) influenza vaccine (CBER-RG8A) in ferrets. To assess cross-reactivity, we generated pseudoviruses bearing HA and NA from circulating A(H5N1) 2.3.4.4b viruses, including North American (B1.13 and D1.1) and Eurasian (DI.2) genotypes. Immunogenicity was assessed using hemagglutination inhibition and microneutralization assays. A single dose elicited robust neutralizing titers (GMT ≥ 160), while a second dose increased titers by ≥3.3-fold. Cross-reactivity was maintained across most strains; however, responses were reduced up to 8-fold against strains harboring the A156T HA mutation, which may introduce a glycosylation site at antigenic site B. Limited responses were detected against divergent clades, with modest titers against clade 2.3.2.1a. These findings suggest broad protection induced by the CSL Seqirus pandemic vaccine against contemporary clade 2.3.4.4b A(H5N1) viruses and underscore the value of ferret immunogenicity data in informing strain selection and regulatory preparedness when human clinical data are unavailable.

Source: 

Link: https://www.nature.com/articles/s41541-026-01438-4

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#Immunity to #hemagglutinin and #neuraminidase results in additive reductions in #airborne #transmission of #influenza #H1N1 virus in #ferrets

 

Abstract

Currently, there is limited knowledge on the impact of immunity to hemagglutinin (HA) and/or neuraminidase (NA) on the transmission of influenza viruses. Therefore, using intramuscular vaccination, intranasal vaccination, or infection with reassortant viruses, we induced immunity to each antigen alone or both antigens combined in ferrets. We then assessed transmission of the 2009 pandemic H1N1 virus from these ferrets to naïve respiratory contacts. For all strategies used to induce immunity, combined immunity to HA and NA resulted in the largest reductions in transmission. Moreover, immunity to HA and NA conferred additive rather than synergistic reductions in transmission. No escape variants emerged in our transmission studies, and logistical regression showed that the probability of transmission was less than 50% when viral titers in donors were reduced to 101.5 and 102 median tissue culture infectious dose per ml on days 1 and 3 postinfection, respectively. These studies define the relationship between immunity to HA and NA on transmission and identify a threshold titer indicative of decreased transmission in ferrets.

Source: 

Link: https://www.science.org/doi/10.1126/sciadv.aea8719

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Defining the transmissible dose 50% for two #pandemic #influenza viruses in #ferrets

 

ABSTRACT

Ferrets are widely used to model airborne transmission of influenza viruses in humans. Airborne transmission is evaluated by infecting donor ferrets with a high virus dose and monitoring transmission to contact animals sharing the same airspace. Humans can be infected with a broad range of influenza virus doses. Therefore, we evaluated the relationship between inoculation dose and transmission for two pandemic influenza viruses in ferrets. Donor ferrets were inoculated with 100 to 106 tissue culture infectious dose 50 (TCID50) of the 2009 pandemic H1N1 or 1968 pandemic H3N2 virus and were then paired with respiratory contacts. Using the proportion of donors that became infected across virus doses, we calculated the infectious dose 50 (ID50). Subsequently, by comparing the proportion of contacts that became infected, we calculated the transmissible dose 50% (TD50): the donor inoculation dose that resulted in transmission to 50% of contacts. For the 2009 pandemic H1N1 virus, the ID50 and TD50 were equivalent at <1 TCID50. However, for the 1968 pandemic H3N2 virus, the ID50 and TD50 were 100.5 and 104.08 TCID50 (95% CI: 102.34–105.82), respectively. The increased TD50 for the H3N2 virus was associated with significant reductions in peak viral titers and viral shedding in donors over decreasing virus inoculation doses. Collectively, these studies define a new measure of transmission that permits comparisons of transmissibility between viral strains and subtypes in ferrets. We show that the 1968 pandemic H3N2 virus has a higher TD50 and reduced transmissibility in ferrets relative to the 2009 pandemic H1N1 virus.

Source: 

Link: https://journals.asm.org/doi/full/10.1128/jvi.01635-25?af=R

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Advancing #H5N1 #influenza #risk #assessment in #ferrets through comparative evaluation of airborne virus shedding patterns

 

Abstract

Recent A(H5N1) zoonotic cases linked to poultry and cattle in North America highlight the urgent need to assess the pandemic potential of emerging strains. Using male ferrets, we evaluate two B3.13 and two D1.1 genotype A(H5N1) viruses isolated from humans and observe fatal disease and varying capacities for direct contact transmission. To enhance pandemic risk assessment, we conduct aerosol sampling using cyclone BC251 and water condensation capture-based SPOT samplers and perform comparative analyses to include additional A(H5N1), A(H9N2), A(H7N9), and A(H1N1)pdm09 strains with known transmissibility profiles. Although none of the A(H5N1) strains transmit via the air, B3.13 viruses are detected at significantly higher levels compared to D1.1 strains. Here we show strong correlations between viral loads in nasal washes, airborne virus shedding, and transmissibility in ferrets, highlighting the value of these metrics for identifying zoonotic influenza viruses that may be adapting toward increased transmission potential.

Source: 

Link: https://www.nature.com/articles/s41467-026-68931-1

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#Oseltamivir and #baloxavir monotherapy and combination #therapy efficacy against clade 2.3.4.4b #H5N1 #influenza virus infection in #ferrets

 

Abstract

Neuraminidase inhibitors (NAIs) and cap-dependent endonuclease inhibitors (CENIs) represent two classes of antiviral drugs recommended for early treatment of patients with seasonal influenza A virus (IAV) infections. However, only limited human data, particularly on combination antiviral treatment, are available to inform optimal dosing regimens against novel IAVs, including highly pathogenic avian influenza A(H5N1) virus, associated with severe disease. Clade 2.3.4.4b A(H5N1) viruses have caused outbreaks in avian and mammalian species worldwide, highlighting the need to assess antiviral drug efficacy against these strains. We challenged ferrets with a D1.1 genotype A(H5N1) virus and treated infected animals with the NAI oseltamivir phosphate (OST) and the CENI baloxavir acid (BXA), alone or in combination, with treatment onset commencing pre- or post-symptom onset (24- or 48-hours post-inoculation (p.i.), respectively). When administered pre- or post-illness onset, BXA, but not OST, monotherapy provided significant reduction of clinical signs and significantly decreased infectious viral levels (in both respiratory and extrapulmonary specimens) compared with mock-treated animals. Combination OST/BXA treatment, when administered pre- or post-symptom onset, resulted in significant improvements in both metrics versus OST monotherapy. These data support continued investigation of antiviral treatment modalities that include both NAI and CENI for patients with mild and severe A(H5N1) disease.

Source: 

Link: https://www.nature.com/articles/s42003-026-09607-w

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Immune history confers #antibody - and T cell-dependent cross-protection against highly pathogenic avian #influenza #H5N1 viruses

 

ABSTRACT

The outbreak of highly pathogenic avian H5 influenza (HPAI) clade 2.3.4.4b in cattle has spread across the United States. Mice with pre-existing immunity to H1N1 virus or with a live-attenuated influenza vaccine showed protection against a lethal bovine-derived HPAI H5N1 viral challenge. Notably, ferrets with mixed immunity also demonstrated protection against a feline-derived H5N1 virus, independent of cross-reactive neutralization titers, but antibodies to whole virus were observed. To investigate protective factors, we conducted T cell epitope mapping using published H1N1 viral sequences and found high conservation of key T cell epitopes in the bovine HPAI H5N1 strain. Depletion of T cells in mice prior to and during primary H1N1 infection impacted cross-protective antibodies to H5N1 virus, with CD4 depletion increasing mortality and CD8 depletion mildly impacting morbidity upon H5N1 viral challenge. This underscores the need to investigate memory T cell responses alongside antibodies in assessing preexisting cross-protection to HPAI H5N1 viruses.

Source: 

Link: https://journals.asm.org/doi/full/10.1128/jvi.02088-25?af=R

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Intranasal #replicon #vaccine establishes mucosal #immunity and protects against #H5N1 and #H7N9 #influenza

 

Abstract

Seasonal and pandemic influenza viruses are continuous threats to human health, requiring rapid development of vaccines to multiple evolving viral strains. RNA vaccine technologies have the adaptability and manufacturability to facilitate pandemic preparedness but have limited flexibility in their route of administration, reducing the ability to establish local protective immune responses such as respiratory mucosal immunity. Here, we describe monovalent and bivalent replicon vaccines against A/Vietnam/1203/2004 H5N1 and A/Anhui/PA-1/2013 H7N9. These replicon vaccines express either H5 or H7 hemagglutinin and are formulated with a nanostructured lipid carrier (NLC) that permits both intramuscular (IM) and intranasal (IN) dosing. In mice, IM vaccination established systemic humoral and cellular responses but no detectable mucosal response, while IN administration induced robust systemic and mucosal immunity. The replicon-NLC vaccines protected against morbidity and mortality in ferret challenge models, establishing this intranasally-administered replicon-NLC vaccine platform as a potential pandemic response tool.

Source: 

Link: https://www.nature.com/articles/s41467-025-64829-6

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Genotype A3 #influenza #H5N1 isolated from fur #seals shows high virulence in #mammals, but not #airborne transmission

 

Abstract

The global spread of highly pathogenic avian influenza A(H5N1) clade 2.3.4.4b viruses has recently extended to include diverse mammalian species, raising new concerns about pandemic risk. In 2023, this clade was first detected in Russian marine mammals during a mass mortality event among northern fur seals in the Far East. Genetic analyses revealed the causative viruses to belong to genotype A3 of European origin, which is known to have circulated in wild birds across the Far East since 2022. Notably, these isolates harbor the mammalian-adaptive substitutions PB2-K482R and NP-N319K—mutations previously linked to enhanced virulence in non-H5 avian influenza viruses, but whose impact on A(H5N1) clade 2.3.4.4b viruses remained to be characterized. The heightened virulence of A3 genotype viruses is confirmed by data obtained via a mouse model. However, despite these adaptive changes, ferret transmission models showed no evidence of airborne transmission of the fur seal-derived virus. Our findings indicate that while PB2-K482R and NP-N319K may contribute to increased mammalian pathogenicity, they do not significantly increase the efficiency of respiratory transmission—a key prerequisite for human pandemic potential. Although suggesting a limited immediate pandemic threat from this A3 genotype, these results underscore the critical need for continued surveillance and functional assessment of emerging mammalian-adaptive mutations in circulating A(H5N1) viruses.

Source: 

Link: https://www.nature.com/articles/s41598-025-28032-3

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#Influenza #mRNA #vaccine reduces #pathogenicity and transmission of #H5N1 virus in a #ferret model

 

Abstract

The global spread of highly pathogenic avian influenza A(H5N1) viruses poses a serious pandemic threat. While sustained human-to-human transmission has not occurred, widespread circulation in birds, increased detection in mammals, and occasional human spillovers underscore the need for safe and effective vaccines. We evaluated an H5 mRNA vaccine candidate in ferrets using recent clade 2.3.4.4b A(H5N1) human isolates. Vaccination elicited strong neutralizing antibodies, conferred robust protection against lethal challenge, and significantly reduced viral titers. In a direct contact transmission model, mRNA vaccination decreased virus shedding in inoculated ferrets and reduced onward transmission; it also protected vaccinated contact ferrets from infection following exposure to virus-shedding, unvaccinated ferrets. Additionally, sera from vaccinated animals cross-neutralized clade 2.3.2.1e human viruses to varying degrees, depending on the strain. These findings demonstrate that H5 mRNA vaccination not only protects against disease but also reduces transmission, supporting its potential as a key tool for pandemic preparedness.

Source: 

Link: https://www.nature.com/articles/s41541-025-01318-3

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#Antigenic Mapping of #H2 #Influenza Viruses recognized by #Ferret and #Human Sera and Predicting Antigenically Significant Sites

 

Abstract

Influenza viruses cause hundreds of thousands of infections globally every year. In the past century, seasonal influenza viruses have included H1N1, H2N2 or H3N2 strains. H2N2 influenza viruses circulated in the human population between 1957-1968. Previously, our group demonstrated a lack of H2N2 influenza virus immunity in individuals born after 1968, as well as the effectiveness of hemagglutinin (HA) based vaccines for multiple influenza virus subtypes. In this study, H2 antigenic maps and radial graphs were generated using previously published data from H2 HA vaccinations of ferrets and seasonal influenza vaccinations of humans. The antigenic maps revealed a stark difference in clustering of HA antigens between the ferrets and humans, and the radial graphs showed specific antigen recognition varies greatly between different influenza preimmune ferrets. These maps also revealed the significant impact that different pre-existing immunities have on antigenic recognition and clustering of antigens after vaccine boost. From these data, we predicted two possible antigenically significant sites containing various mutations that have not been previously reported and showed that one of these sites relevant using mouse anti-sera.

Source: BioRxIV, https://www.biorxiv.org/content/10.1101/2025.11.12.687988v1

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Intramammary #infection of bovine #H5N1 #influenza virus in #ferrets leads to transmission in suckling neonates

 

Abstract

The spread of H5N1 clade 2.3.4.4b in dairy herds raises concerns about zoonotic transmission due to its high viral load in milk, a key contact point between livestock and humans. H5N1 clade 2.3.4.4b exhibits tropism for the mammary gland, with milk from infected animals containing high levels of infectious virus, posing potential risks to offspring via breastfeeding. Using a lactating ferret model, we demonstrate that mammary gland infection with bovine H5N1 transmits the virus to suckling kits, resulting in neonatal mortality. Viral RNA levels increased in milk and remained high in mammary tissue, with infected kits exhibiting elevated viral RNA in the oral and nasal cavities and feces. Additionally, we detected the H5N1 receptor, α2,3 sialic acid, in ferret and human mammary tissue. These data demonstrate that H5N1 clade 2.3.4.4b infection in lactating dams leads to mastitis-related disease and transmits to suckling pups, resulting in mortality among neonates.

Competing Interest Statement

The authors have declared no competing interest.

Source: BioRxIV, https://www.biorxiv.org/content/10.1101/2024.11.15.623885v3

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#Transmission of #SARS-CoV-2 between #ferrets in presence of pre-existing #immunity

 

ABSTRACT

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the etiological agent of coronavirus disease 2019 (COVID-19), is characterized by its high contagiousness. The COVID-19 pandemic has exerted profound impacts on human society. The persistent circulation of SARS-CoV-2 in human populations continues to pose re-exposure risks for both vaccinated individuals and those with prior natural infection. Against this epidemiological background, there is an urgent need to characterize the transmission dynamics of SARS-CoV-2 in the context of pre-existing immunity. Using a ferret infection model, this study systematically addresses critical scientific questions, including viral transmission efficiency, temporal patterns of transmissibility, and the ability of pre-existing immunity to mitigate reinfection and viral shedding. The findings provide robust experimental evidence for elucidating the transmission mechanisms of SARS-CoV-2 and offer scientific insights to inform the rational design of optimized antiviral strategies.

Source: Journal of Virology, https://journals.asm.org/doi/full/10.1128/jvi.01566-25?af=R

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Post-infection #pig and #ferret antisera show similar #antigenic profiles for #human #influenza #H1N1pdm09 viruses

 

Abstract

Background: 

Monitoring antigenic drift in human influenza A viruses is essential for vaccine strain selection and ensuring protection against circulating strains. Antigenic drift is traditionally assessed using ferret antisera, which provide monospecific responses, and human vaccinee sera, which reflect exposure to multiple antigens. In this study we evaluated the pig as an alternative source of antisera to study antigenic drift compared to immune responses in ferrets and humans. We included seasonal influenza A(H1N1pdm09) human viruses that had shown different antigenic characteristics when using ferret or human antisera. 

Methods: 

Pairs of pigs were inoculated with six human A(H1N1)pdm09 viruses circulating between 2019 and 2023, a period of marked antigenic drift. Pig and ferret antisera were analysed by hemagglutination inhibition (HI) and virus neutralization (VN) assays. 

Results: 

Pigs were successfully infected with all strains, shedding virus and producing antibody responses, confirming their susceptibility to human influenza A viruses. Antigenic reactivity of pig antisera was qualitatively comparable to ferret antisera in both HI and VN assays, although maximum homologous antibody titres were significantly higher in ferrets. The antisera raised against viruses in circulation in 2019 and before, exempified by A/Guangdong-Maonan/SWL1536/2019, clade 5a.1, were clearly differentiated by both ferret and pig antisera from those in clade 5a.2 and its derivatives that became predominant. 

Conclusions: 

Ferrets and pigs showed comparable responses and both distinguished clade 5a.1 from clade 5a.2. However, neither model recognised antigenically drifted variants from 2019/2022, including subclades 5a.2 C, 5a.2a C.1/C.1.9, and .5a.2a.1 C.1.1/D, which were distinguishable using human post-vaccination antisera.

Competing Interest Statement

The authors have declared no competing interest.

Funder Information Declared

Biotechnology and Biological Sciences Research Council, https://ror.org/00cwqg982, BB/X511134/1 , BBS/E/PI/230002A, BBS/E/PI/230002B, BBS/E/PI/23NB0004, BBS/E/PI/23NB0003, BB/Y007298/1

Medical Research Council, https://ror.org/03x94j517, CC1114

Cancer Research UK, CC1114

Source: BioRxIV, https://www.biorxiv.org/content/10.1101/2025.10.30.685573v1

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A #vaccine central in #H5 #influenza antigenic space confers broad #immunity

 

Abstract

Highly pathogenic avian influenza A(H5) viruses globally impact wild and domestic birds, and have caused severe infections in mammals, including humans, underscoring their pandemic potential. The antigenic evolution of the A(H5) haemagglutinin (HA) poses challenges for pandemic preparedness and vaccine design. Here the global antigenic evolution of the A(H5) HA was captured in a high-resolution antigenic map. The map was used to design immunogenic and antigenically central vaccine HA antigens, eliciting antibody responses that broadly cover the A(H5) antigenic space. In ferrets, a central antigen protected as well as homologous vaccines against heterologous infection with two antigenically distinct viruses. This work showcases the rational design of subtype-wide influenza A(H5) pre-pandemic vaccines and demonstrates the value of antigenic maps for the evaluation of vaccine-induced immune responses through antibody profiles.

Source: Nature, https://www.nature.com/articles/s41586-025-09626-3

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Protective efficacy of the UniFluVec #influenza #vaccine vector against the highly pathogenic influenza A/Indonesia/5/2005 #H5N1 strain in #ferrets

 

Highlights

• UniFluVec, an H1N1pdm vaccine candidate, includes NS1 and NEP modifications to boost attenuation and immunity.

• UniFluVec protects ferrets from H5N1, enhancing clearance, limiting lung damage, and ensuring 100 % survival after one dose.

• Replication-deficient UniFluVec shows cross-protection, supporting its potential as a pre-pandemic intranasal vaccine.

Abstract

Background

The emergence of new influenza strains with unpredictable antigenic properties poses a significant vaccination challenge. The increasing incidence of human H5 infections underscores the urgent need for effective pre-pandemic vaccines.

Methods

The UniFluVec and UniFluVec-wtNS1 viruses were designed as H1N1pdm vaccine candidates. Both viruses contained a heterologous A/Singapore/1/57-like (H2N2) NEP gene, which served as an attenuation factor. UniFluVec additionally carried a truncated to 124 amino acids NS1 gene, and an insertion of conserved influenza sequences. UniFluVec-wtNS1 retained the wild-type NS1 gene. The impact of NS1 and NEP modifications on attenuation and phenotypic markers was assessed in cells and mice. Safety and prophylactic efficacy were assessed in ferrets following a single intranasal immunisation with the maximum feasible dose (8.7 log10 EID50), followed by challenge with the highly pathogenic avian influenza virus (HPAIV) A/Indonesia/5/2005 (H5N1).

Results

Modifications in NS1 and NEP independently and synergistically induced a temperature-sensitive phenotype and enhanced type I/II interferon response, resulting in a highly attenuated vaccine profile. UniFluVec, incorporating both modifications within the NS genomic segment, demonstrated superior viral clearance, reducing lung damage, and ensuring 100 % survival in infected animals.

Conclusion

The replication-deficient UniFluVec vector demonstrates safety, immunogenicity, and protective efficacy against the heterologous HPAIV strain in ferrets following a single intranasal administration.

Source: Vaccine, https://www.sciencedirect.com/science/article/pii/S0264410X25010928?via%3Dihub

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Identification of #clinical and #virological correlates associated with #influenza A candidate #vaccine virus (#CVV) attenuation in a #ferret model

 

ABSTRACT

Influenza A viruses continuously circulate among avian and swine species, posing a persistent threat to public health. The development of influenza candidate vaccine viruses (CVVs) plays a pivotal role in the global strategy for influenza pandemic preparedness. Safety-testing of CVVs for attenuation in ferrets represents a critical step that takes place prior to making these viruses available to vaccine manufacturers. Development of pathogenicity standards is needed to establish acceptable thresholds of disease so that CVV safety can be assessed without the need for comparison to the parental virus. To assess the capacity of diverse CVVs to cause pathogenesis in mammalian hosts, clinical and virological parameters were compiled from CVV assessments in ferrets conducted using consistent methods over approximately 20 years to identify disease parameters most reflective of attenuation compared to wild-type strains. These analyses revealed an overall reduction in ferret weight loss and fever relative to wild-type controls. Viral titers in nasal washes were reduced with limited spread to tissues beyond the respiratory tract. Regression models further support the significance of clinical signs in distinguishing the virulence of wild-type viruses and CVVs. These findings provide support for the development of standardized parameters for assessing pathogenicity of CVVs and their suitability for manufacturers.

IMPORTANCE

The development and safety testing of candidate vaccine viruses (CVVs) against emerging zoonotic influenza strains prior to sharing with vaccine manufacturers is a critical component of influenza pandemic preparedness. The extensive data set reported here provides critical information that will drastically streamline the safety testing process, thereby enabling more efficient CVV assessments and improving public health in the event of an influenza pandemic.

Source: Journal of Virology, https://journals.asm.org/doi/full/10.1128/jvi.01023-25?af=R

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Effect of Seasonal #Influenza #Vaccines on Avian Influenza #H5N1 Clade 2.3.4.4b Virus #Infection in #Ferrets

 

Abstract

Highly pathogenic avian influenza A(H5N1) clade 2.3.4.4b viruses have infected >1,000 herds of dairy cattle and hundreds of poultry flocks in the United States since the beginning of 2024. Seventy human cases have been reported during that period, mainly through occupational exposure. Although prior influenza A(H1N1)pdm09 virus infection has been shown to confer protection against influenza A(H5N1) clade 2.3.4.4b virus infection in the ferret model, it remains unclear if influenza vaccines, known to elicit a less potent and narrower cross-reactive immune response, can achieve a similar effect. In this article, we demonstrate that immunization with commercially available human seasonal influenza vaccines also confers partial protection against disease caused by H5N1 clade 2.3.4.4b virus in ferrets, which is partially associated with the presence of cross-reactive antibodies targeting H5N1 virus antigens.

Source: US Centers for Disease Control and Prevention, https://wwwnc.cdc.gov/eid/article/31/10/25-0668_article

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