ETIDIOH - NEW

  Highlights •  Concern over H5N1 bird flu testing and detection in the Netherlands is increasing. •  50 human laboratories in the Netherlands, Aruba, Bonaire , and Curacao were assessed. •  The laboratories detected animal influenza viruses with high performance. •  Few laboratories identified the animal subtype of detected influenza A viruses. •  National reference laboratory capacity to identify the animal subtype is critical. Abstract Background Since 2022, highly pathogenic H5N1 influenza A virus clade 2.3.4.4b has caused global outbreaks among wild birds and poultry , with increasing mammalian and sporadic human infections . This elevates concerns about zoonotic transmission and pandemic risk , highlighting the need for accurate detection and identification of animal influenza A viruses by human clinical diagnostic laboratories (hCDL). Methods To evaluate routine diagnostic performance , an External Quality Assessment (EQA) panel containing inactivate...

  Significance Middle East respiratory syndrome coronavirus (MERS-CoV) remains the most lethal human coronavirus , with continued zoonotic transmission from wild naturally infected dromedary camels , posing a persistent risk of spillover to humans. Despite this ongoing threat , no specific antiviral treatment has been approved. In this study, we characterize the antibody response to MERS-CoV in naturally infected dromedaries, the primary animal reservoir, and identify a panel of nanobodies (Nbs) exhibiting potent neutralizing activity . These Nbs recognize a previously unreported binding and neutralizing site on the virus spike receptor-binding domain (RBD). Their distinctive genetic, structural, and functional properties make them promising candidates for the development of effective and therapeutic interventions against MERS-CoV, as strongly advocated by global health authorities. Abstract Wild dromedary camels in the Arabian Peninsula and Africa have harbored antibodies against ...

  Abstract Influenza (flu) is a common respiratory virus with seasonal global spread. Zoonotic viruses can occasionally cross species, leading to pandemic-level spread , and for flu viruses, this is considered an “antigenic shift ”. The flu can be particularly severe during pregnancy due to immune system adaptations that occur during pregnancy, with prior global pandemics causing excess hospitalizations, deaths, and other complications in the mothers and the neonates. We aim to review the current literature with respect to novel avian H5N1 and the potential impact of infection with flu during pregnancy. A systematic literature search was conducted. Here we provide a rapid summary of epidemiology and understanding of viral spread , published risks of H5N1 in pregnancy, the unique physiologic, cellular, and molecular adaptations making H5N1 infection unique in pregnancy, implementation of an effective vaccine program in event of a pandemic specific to pregnant individuals, optimizing...

  Abstract Middle East respiratory syndrome coronavirus (MERS-CoV) was identified as a human pathogen in 2012 and causes ongoing sporadic infections and outbreak clusters . Despite case fatality rates (CFRs) of over 30% and considerable pandemic potential , a safe and efficacious vaccine has not been developed. Here we report the design, characterization, and preclinical evaluation of MERS-CoV antigens . Our lead candidate comprises a stabilized spike displayed on a self-assembling ferritin nanoparticle that can be produced from a high-expressing, stable cell pool . This vaccine elicits robust MERS-CoV pseudovirus and authentic virus neutralizing antibody titers in BALB/c mice. Immunization of male non-human primates (NHPs) with one dose of Alhydrogel-adjuvanted vaccine elicited a > 103 geometric mean titer of pseudovirus neutralizing antibodies that was boosted with a second dose. Sera from these NHPs exhibited cross-reactivity against spike-pseudotyped lentiviruses from MERS-C...

  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 ...

  Highlights •  IN-delivered ChAd-Texas vaccine elicits mucosal antibody and T cell responses •  IN-delivered ChAd-Texas vaccine protects against H5N1 in mice and hamsters •  IN delivery of ChAd-Texas vaccine confers greater protection than IM delivery •  ChAd-Texas induces H5N1 immunity in the setting of prior influenza immunity Summary The emergence of highly pathogenic avian H5N1 influenza viruses in dairy cows and humans has increased the potential for another pandemic . To address this risk, we developed chimpanzee adenoviral (ChAd)-vectored H5 hemagglutinin-targeted vaccines and tested their immunogenicity and efficacy in rodents . Immunization with ChAd-Texas (clade 2.3.4.4b) vaccine in mice elicits neutralizing antibody responses and confers protection against viral infection and mortality upon challenge with a human H5N1 isolate (A/Michigan/90/2024, clade 2.3.4.4b). Intranasal delivery of the ChAd-Texas vaccine elicits mucosal antibody and T cell respon...

  Abstract Influenza A viruses continually pose pandemic threats, underscoring the need for timely development of Candidate Vaccine Viruses (CVVs) that meet regulatory expectations for vaccine manufacturing. This protocol describes the procedures used at CDC to generate recombinant CVVs through reverse genetics in accordance with World Health Organization guidelines and CDC’s internal Quality System Requirements (QSR)1,2,3. The QSR incorporates relevant principles from the FDA’s Good Manufacturing Practice (GMP) and Good Laboratory Practice (GLP) regulations, providing a structured framework that ensures documentation integrity, material traceability, and quality oversight during all stages of CVV development. The protocol provides detailed steps for plasmid preparation , virus rescue in Vero cells , and amplification in embryonated chicken eggs , and outlines characterization assays used to confirm the suitability and safety attributes of each CVV. This standardized, quality-drive...

  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 on...

  Abstract The continuous evolution and global spread of highly pathogenic avian influenza (HPAI) H5N1 viruses , particularly clade 2.3.4.4b, pose major challenges for pandemic preparedness . This study evaluates a low-dose inactivated split-virus vaccine derived from H5N1 clade 2.3.4.4b, formulated with an Alum/CpG adjuvant , in a preclinical female mouse model . The vaccine induces strong humoral and cellular immunity , generating high titers of cross-reactive antibodies against diverse H5 hemagglutinin (HA) and across different N1 neuraminidase (NA) glycoproteins. The Alum/CpG adjuvant supports substantial antigen dose sparing and promotes a balanced Th1/Th2 profile. Functional assays show potent virus neutralization , neuraminidase inhibition , and antibody-dependent cellular cytotoxicity , alongside robust antigen-specific CD4+ and CD8+ T cell responses, efficient control of lung viral replication, and reduced lung inflammation . Vaccinated mice are fully protected from lethal...

  Abstract The risk of highly pathogenic avian influenza infection to humans is challenging to estimate because many human avian influenza virus (AIV) infections are undetected as they may be asymptomatic , symptomatic but not tested , and as contact tracing is difficult because human-to-human spread is rare. We derive equations that consider the evolutionary mechanisms that give rise to pandemics and are parameterized to be consistent with records of past pandemics. We estimate that thousands of human AIV infections occur worldwide in an average year and estimate the infection fatality ratio as 32 deaths per 10,000 infections (95% confidence interval: [9.6, 75]). We estimate that preventing 20% of animal-to-human influenza spillovers annually would delay pandemic emergence by an average of 9.4 years . There is a high level of uncertainty in our estimates due to the few records of past pandemics, but even so this infection fatality ratio is comparable to SARS-CoV-2 during the recen...

  Abstract We evaluated 2 electrochemiluminescence serologic assays to detect avian influenza H5 antibodies . Both assays identified H5 antibodies from both serum and dried blood spots and had strong specificity and minimal cross-reactivity in human and avian samples. Such assays can support populationwide serologic surveys aimed at assessing population-level immunity. Source:  Link:  https://wwwnc.cdc.gov/eid/article/32/1/25-1117_article ____

  Abstract In 2009 and again in 2019, public health warnings were confirmed by the emergence, rapid widespread transmission, and lethality of novel influenza and coronaviruses . The world continues to suffer disease from these respiratory viruses. Two newly recognized emergent respiratory viruses, influenza D and canine coronavirus HuPn-2018 , have been shown to have considerable potential for causing future human epidemics , but diagnostics and surveillance for the viruses are lacking. We reviewed data regarding influenza D virus and coronavirus canine coronavirus HuPn-2018 . Those data strongly indicate that these viruses are major newly recognized threats . However, little is being done to respond to or prevent disease associated with these viruses, warranting the question of whether we will learn from previous pandemics. Source:  Link:  https://wwwnc.cdc.gov/eid/article/32/1/25-1764_article ____

Abstract Highly pathogenic avian influenza A (H5N1) (H5N1 hereafter) is an emerging pathogen in mammals . The recent recognition of H5N1 in dairy cattle increases opportunities for human exposure and infection and may accelerate a trajectory toward sustained human-to-human transmission . Furthermore, the presence of virus at high concentration in unpasteurized milk raises new risks for humans , especially infants and children . Milk has been identified as a vehicle for viral transmission in and between mammalian species, including humans. Sialic acids (SAs) found on cell surfaces are important mediators of species susceptibility to specific influenza strains and play an important role in viral tropism . New data demonstrate that SA receptors with α2,3 linkages capable of binding avian influenza strains are present in human mammary tissue . The presence of SA receptors that can bind avian influenza and a comparative analysis of viral transmission risk of raw and pasteurized milk in seve...

  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 m...

  Abstract Highly pathogenic H5N1 avian influenza viruses of clade 2.3.4.4b cause sporadic human infections and currently raise concerns about a new influenza pandemic . Heterogeneities in disease severity have been observed in the past and are reported among infected farm workers in the United States . These may be attributed to differences in pre-existing H5N1 cross-reactive antibodies . In this study, we characterize H5N1 cross-reactive antibody landscapes in the current population (#NCT05794412 and #NCT01022905) and assess the effect of AS03-adjuvanted pandemic H1N1 and non-adjuvanted seasonal influenza vaccination on H5N1 cross-neutralizing and IgG antibody titers targeting a range of influenza virus-derived antigens. We detect H5N1 cross-neutralizing antibodies using a vesicular stomatitis virus-based pseudovirus system that correlate well with antibodies inhibiting the spread of authentic H5N1 viruses, anti-group 1 hemagglutinin stalk and anti-trimeric hemagglutinin antibodi...

  {Abstract} Background :  Emerging threats such as highly pathogenic influenza strains like H5N1 emphasize the need for vaccines that induce cross-reactive immunity against conserved epitopes. Existing influenza vaccines primarily elicit strain-specific responses , leaving gaps in protection against pandemic subtypes. This study aimed to evaluate T cell responses to seasonal influenza A and H5N1 and compare them to SARS-CoV-2 specific T cell responses to understand differences shaped by distinct exposure histories and vaccination strategies. Methods :  T cell responses were assessed in 41 laboratory workers who received annual seasonal influenza vaccines using ELISpot to quantify responses to peptide pools derived from influenza ( H1N1 hemagglutinin [HA], H3N2 HA, H5N1 HA, matrix protein 1 [MP1], nucleoprotein [NP]) and SARS-CoV-2 (spike [S2S], nucleocapsid [S2N]). Ten-day expansion assays were used to evaluate functional cross-reactivity between H1, H3, and H5 HA. Intra...

  Abstract Avian influenza viruses (AIVs), including H5N1 and H7N9, from the Orthomyxoviridae family present substantial public health concerns . The predominant circulating clade 2.3.4.4b has demonstrated enhanced capacity for mammalian adaptation , raising concerns about potential reassortment with human seasonal influenza viruses . Unlike H7N9 ’s limited host range , H5N1 infects birds, various mammals, and humans . Recent concerns include widespread H5N1 infection of U.S. dairy cattle across 18 states , affecting over 1000 herds with 71 human infections (70 H5N1 and 1 H5N5). Key observations include cow-to-cow transmission, viral presence in milk, and transmission to humans , mainly through occupational exposure . Evidence of mammal-to-mammal transmission has been documented in European and Canadian foxes and South American marine mammals . Standard pasteurization effectively inactivates the virus in milk. The continuing mammalian adaptations , particularly mutations like PB2-E...

  Abstract Avian influenza is a global public health threat. Since 2021, the ongoing H5N1 panzootic has brought a major shift in H5Nx epidemiology , including unprecedented spread, wide host range and lack of seasonality . Infections in marine mammals, wildlife and livestock have heightened concern for human-to-human transmission and pandemic potential . Contact tracing and self-isolation are used as public health measures in the UK to manage contacts of confirmed human cases of avian influenza. In this study, we aimed to estimate potential outbreak sizes and evaluate the effectiveness of contact tracing and self-isolation in managing community outbreaks of H5N1 following spillover from birds to people . We characterised contact patterns from an underrepresented agricultural population at high risk of avian influenza exposure through contact with birds (Avian Contact Study). Informed by these realistic social contact data, we modelled outbreak sizes using a stochastic branching pro...

  Abstract Human (avian) influenza A viruses, especially highly pathogenic avian influenza (HPAI) viruses, pose a significant public health threat , and a multivalent vaccine is the primary prophylactic measure to control these viruses. To establish such a vaccine, we generated two multivalent vesicular stomatitis virus (VSV)-based vaccine candidates (V-EtM2e/H505 and V-EtM2e/H522) and characterized their ability to induce protective immune responses. Our results revealed that vaccine immunization in mice induced high humoral immune responses against both the HPAI hemagglutinin (HA) protein and the ectodomain of M2 (M2e) protein . Intriguingly, vaccine-immunized mouse sera exhibited highly efficient neutralizing activity against the corresponding H5 pseudovirus and mediated potent and broad antibody-dependent cellular cytotoxicity (ADCC) activity against M2e derived from human and avian influenza H5, H1, H3, and H7 viruses . Furthermore, both intranasal and intramuscular immunizati...

  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...