Neuraminidase Inhibitors Zanamivir Research Articles

Low antiviral resistance in Influenza A and B viruses isolated in Mexico from 2010 to 2023

The most widely used class of antivirals available for Influenza treatment are the neuraminidase inhibitors (NAI) Oseltamivir and Zanamivir. However, amino acid (AA) substitutions in the neuraminidase may cause reduced inhibition or high antiviral resistance. In Mexico, the current state of knowledge about NAI susceptibility is scarce, in this study we report the results of 14 years of Influenza surveillance by phenotypic and genotypic methods. A total of 255 isolates were assessed with the NAI assay, including Influenza A(H1N1)pdm09, A(H3N2) and Influenza B (IBV). Furthermore, 827 sequences contained in the GISAID platform were analyzed in search of relevant mutations.Overall, five isolates showed highly reduced inhibition or reduced inhibition to Oseltamivir, and two showed reduced inhibition to Zanamivir in the NAI assays. Additionally, five A(H1N1)pdm09 sequences from the GISAID possessed AA substitutions associated to reduced inhibition to Oseltamivir and none to Zanamivir. Oseltamivir resistant A(H1N1)pdm09 harbored the H275Y mutation. No genetic mutations were identified in Influenza A(H3N2) and IBV. Overall, these results show that in Mexico the rate of NAI resistance is low (0.6%), but it is essential to continue the Influenza surveillance in order to understand the drug susceptibility of circulating strains.

Heteromultivalent Ligand Display on Reversible Self-Assembled Monolayers (rSAMs): A Fluidic Platform for Tunable Influenza Virus Recognition.

We report on the design of heteromultivalent influenza A virus (IAV) receptors based on reversible self-assembled monolayers (SAMs) featuring two distinct mobile ligands. The principal layer building blocks consist of α-(4-amidinophenoxy)alkanes decorated at the ω-position with sialic acid (SA) and the neuraminidase inhibitor Zanamivir (Zan), acting as two mobile ligands binding to the complementary receptors hemagglutinin (HA) and neuraminidase (NA) on the virus surface. From ternary amphiphile mixtures comprising these ligands, the amidines spontaneously self-assemble on top of carboxylic acid-terminated SAMs to form reversible mixed monolayers (rSAMs) that are easily tunable with respect to the ligand ratio. We show that this results in the ability to construct surfaces featuring a very strong affinity for the surface proteins and specific virus subtypes. Hence, an rSAM prepared from solutions containing 15% SA and 10% Zan showed an exceptionally high affinity and selectivity for the avian IAV H7N9 (Kd = 11 fM) that strongly exceeded the affinity for other subtypes (H3N2, H5N1, H1N1). Changing the SA/Zan ratio resulted in changes in the relative preference between the four tested subtypes, suggesting this to be a key parameter for rapid adjustments of both virus affinity and selectivity.

New insights into the neuraminidase-mediated hemagglutination activity of influenza A(H3N2) viruses

Influenza virus neuraminidase (NA) can act as a receptor-binding protein, a role commonly attributed to hemagglutinin (HA). In influenza A(H3N2) viruses, three NA amino acid residues have previously been associated with NA-mediated hemagglutination: T148, D151, and more recently, H150. These residues are part of the 150-loop of the NA monomer. Substitutions at 148 and 151 arise from virus propagation in laboratory cell cultures, whereas changes at 150 occurred during virus evolution in the human host. In this study, we examined the effect of natural amino acid polymorphism at position 150 on NA-mediated hemagglutination. Using the A/Puerto Rico/8/34 backbone, we generated a comprehensive panel of recombinant A(H3N2) viruses that have different NAs but shared an HA that displays poor binding to red blood cells (RBCs). None of the tested substitutions at 150 (C, H, L, R, and S) promoted NA-binding. However, we identified two new determinants of NA-binding, Q136K and T439R, that emerged during virus culturing. Similar to T148I, both Q136K and T439R reduced NA enzyme activity by 48–86% and inhibition (14- to 173-fold) by the NA inhibitor zanamivir. NA-binding was observed when a virus preparation contained approximately 10% of NA variants with either T148I or T439R, highlighting the benefit of using deep sequencing in virus characterization. Taken together, our findings provide new insights into the molecular mechanisms underlying the ability of NA to function as a binding protein. Information gained may aid in the design of new and improved NA-targeting antivirals.

An armed anti-immunoglobulin light chain nanobody protects mice against influenza A and B infections.

The immune system eliminates pathogen intruders such as viruses and bacteria. To recruit immune effectors to virus-infected cells, we conjugated a small molecule, the influenza neuraminidase inhibitor zanamivir, to a nanobody that recognizes the kappa light chains of mouse immunoglobulins. This adduct was designed to achieve half-life extension of zanamivir through complex formation with the much-larger immunoglobulins in the circulation. The zanamivir moiety targets the adduct to virus-infected cells, whereas the anti-kappa component simultaneously delivers polyclonal immunoglobulins of indeterminate specificity and all isotypes. Activation of antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity promoted elimination of influenza neuraminidase-positive cells. A single dose of the conjugate protected mice against influenza A or B viruses and was effective even when given several days after infection with a lethal dose of virus. In the absence of circulating immunoglobulins, we observed no in vivo protection from the adduct. The type of conjugates described here may thus find application for both anti-influenza prophylaxis and therapy.

Zanamivir infusion improves endothelial function and reduces arterial stiffness in diabetic mice

Endothelial dysfunction and arterial stiffening are consistently manifested in type 2 diabetes (T2D) and contribute to the development of cardiovascular disease. Consequently, there is an urgent need to identify novel therapies that improve endothelial function and reduce arterial stiffness in T2D. Endothelial dysfunction is characterized by a reduced capacity of the endothelium to induce vasodilation. Physiologically, this is evident as an impairment in flow-mediated dilation, a process in which the endothelial glycocalyx participates in the mechanosensation and mechanotransduction of shear that leads to the production of the vasodilator nitric oxide. Nitric oxide, in turn, has also been shown to be a potent regulator of vascular wall homeostasis, including arterial stiffness. In T2D, circulating neuraminidase activity is increased and implicated in glycocalyx degradation and endothelial dysfunction. Accordingly, we hypothesized that inhibition of neuraminidase activity would enhance endothelial function and reduce arterial stiffness in T2D. We tested this hypothesis in diabetic db/db mice infused with zanamivir (n=10) or vehicle control (n=10) for 4 weeks via subcutaneously implanted osmotic mini pumps. We chose to use the FDA approved neuraminidase inhibitor zanamivir to increase the translatability of our study. Treatment with zanamivir did not affect body weight or blood pressure. Nonetheless, it improved flow-mediated dilation in isolated mesenteric arteries assessed using pressure myography, with no associated changes in endothelium-independent vasodilation. Zanamivir also reduced stiffness in isolated mesenteric and femoral arteries, as well as aortic stiffness as assessed in vivo via pulse wave velocity and ex vivo via atomic force microscopy. These results indicate that zanamivir is a potential therapeutic to ameliorate vascular dysfunction in T2D. National Institutes of Health grant: R01HL153264 to LM-L and JP This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Report on influenza viruses received and tested by the Melbourne WHO Collaborating Centre for Reference and Research on Influenza during 2022.

As part of its role in the World Health Organization's (WHO) Global Influenza Surveillance and Response System (GISRS), the WHO Collaborating Centre for Reference and Research on Influenza in Melbourne received a record total of 12,073 human influenza positive samples during 2022. Viruses were analysed for their antigenic, genetic and antiviral susceptibility properties. Selected viruses were propagated in qualified cells or embryonated hen's eggs for potential use in seasonal influenza virus vaccines. In 2022, influenza A(H3N2) viruses predominated over influenza A(H1N1)pdm09 and B viruses, accounting for 77% of all viruses analysed. The majority of A(H1N1)pdm09, A(H3N2) and influenza B viruses analysed at the Centre were found to be antigenically and genetically similar to the respective WHO recommended vaccine strains for the southern hemisphere in 2022. Of 3,372 samples tested for susceptibility to the neuraminidase inhibitors oseltamivir and zanamivir, two A(H1N1)pdm09 viruses showed highly reduced inhibition against oseltamivir.

Diagnosis and treatment of influenza: a literature review

Alisa Pozniak1 1Vilnius University, Faculty of Medicine, Vilnius, Lithuania Abstract Background. Influenza is an acute viral respiratory infection. Influenza viruses spread through respiratory droplets and are characterized by frequent mutations that lead to antigenicity changes. According to the World Health Organization, seasonal influenza epidemics cause approximately 4 million cases of severe illness and 250,000 to 500,000 deaths each year globally. Aim: to review the diagnosis and treatment of influenza. Methodology: literature sources were selected from PubMed database following dates from 2017 to 2022 using keywords and their combinations: influenza, rapid antigen test, neuraminidase inhibitors, favipiravir, baloxavir. Results. Influenza should be suspected in patients who develop acute respiratory symptoms during the influenza season. Diagnostic tests available for influenza include rapid antigen and molecular tests, polymerase chain reaction tests, virological culture. Antiviral therapy should be initiated in patients who are at increased risk of complications from influenza. The neuraminidase inhibitors zanamivir and oseltamivir are used for specific antiviral therapy. New antivirals such as peramivir, favipiravir, laninamivir and baloxavir have emerged as a promising new treatment approach. Conclusions. Influenza is a serious infectious disease that can lead to life-threatening complications. Virological diagnosis is based on specific epidemiological data, clinical symptoms and laboratory tests. Neuraminidase inhibitors are recommended for early treatment. Several cases of zanamivir and oseltamivir-resistant virus have been reported. That is the reason why different approaches for antiviral treatment are being investigated. Keywords: influenza, rapid antigen test, neuraminidase inhibitors, favipiravir, baloxavir.

Antiviral Susceptibilities of Avian Influenza A(H5), A(H7), and A(H9) Viruses Isolated in Japan

The circulation of avian influenza A viruses in poultry is a public health concern due to the potential transmissibility and severity of these viral infections. Monitoring the susceptibility of these viruses to antivirals is important for developing measures to strengthen the level of preparedness against influenza pandemics. However, drug susceptibility information on these viruses is limited. Here, we determined the susceptibilities of avian influenza A(H5N1), A(H5N2), A(H5N8), A(H7N7), A(H7N9), A(H9N1), and A(H9N2) viruses isolated in Japan to the antivirals approved for use there: an M2 inhibitor (amantadine), neuraminidase inhibitors (oseltamivir, peramivir, zanamivir, and laninamivir) and RNA polymerase inhibitors (baloxavir and favipiravir). Genotypic methods that detect amino acid substitutions associated with antiviral resistance and phenotypic methods that assess phenotypic viral susceptibility to drugs have revealed that these avian influenza A viruses are susceptible to neuraminidase and RNA polymerase inhibitors. These results suggest that neuraminidase and RNA polymerase inhibitors currently approved in Japan could be a treatment option against influenza A virus infections in humans.

Drug-induced Neuropsychiatric Adverse Events Using Post-Marketing Surveillance.

Several studies reported that abnormal behavior was noted in pediatric patients receiving several drugs, including neuraminidase inhibitors (NIs). However, the information on drugs associated with abnormal behavior in a real-world setting remains limited. The purpose of this study was to clarify the drugs associated with abnormal behavior using a spontaneous reporting system database. We performed a retrospective pharmacovigilance disproportionality analysis using the Japanese Adverse Drug Event Report database. Adverse event reports submitted to the Pharmaceuticals and Medical Devices Agency were analyzed, and the reporting odds ratio at 95% confidence interval were calculated. A total of 1,144 reports of abnormal behavior were identified. The signals were detected through the association of 4 neuraminidase inhibitors (oseltamivir, zanamivir, laninamivir, and peramivir) with the abnormal behaviour. These signals were stronger for oseltamivir than other neuraminidase inhibitors. The signals were also detected for acetaminophen and montelukast. Our results should be able to raise physicians' awareness of drugs associated with abnormal behavior, but further investigation of these medications is warranted.

Spectroscopic, electrochemical, and some theoretical studies on the interactional of neuraminidase inhibitor zanamivir with double helix deoxyribonucleic acid

In some cases, instead of designing new molecules, investigating the potential of molecules with small chemical structures with biological activity to interact with double-stranded DNA is very important for the design of drug candidate molecules that can target new DNA. In our literature search, we did not encounter the mode of binding of Zanamivir (ZAN), an antiviral drug, with its double-stranded DNA structure. For this reason, the types of interactions between dsDNA and a ZAN were investigated experimentally and theoretically by comparative experimental and theoretical studies between different groups. This study covers spectroscopic studies such as absorption, fluorescence, and FT-IR, electrochemical properties such as cyclic voltammetry, and some physical properties such as viscosity and theoretical studies at certain pH values. As a result of our theoretical studies (according to the results obtained from the lowest energy insertion poses) supporting our experimental studies, we concluded that ZAN prefers the minor groove binding mechanism and settles into the double helix structure.

Characterization of influenza B viruses with reduced susceptibility to influenza neuraminidase inhibitors

A total of 3425 influenza B viruses collected from the Asia-Pacific region were tested against the four registered neuraminidase inhibitors (NAIs) (oseltamivir carboxylate, zanamivir, peramivir and laninamivir) as part of the routine surveillance work at the WHO Collaborating Centre for Research and Reference on Influenza, Melbourne between 2016 and 2020. Forty-five influenza B viruses with reduced susceptibility to one or more NAIs were identified. While the majority of these had neuraminidase (NA) mutations that were known to confer NAIs resistance, fifteen had NA mutations that had not been confirmed as being responsible for reduced NAIs susceptibility. Eleven of these NA mutations of concern were investigated using reverse genetics (RG) techniques to verify that these mutations were the cause of the reduced NAI susceptibility. All mutations were introduced separately into the NA of B/Brisbane/27/2016 (a B Victoria-lineage virus) or B/Yamanashi/166/98 (a B Yamagata-lineage virus) and the effects of these were analysed by an in vitro NAI assay. The T146K substitution in the NA of B Victoria and Yamagata-lineages resulted in a large increase in the IC50 for peramivir (>1000-fold increase in the mean IC50 of sensitive viruses with T146) with smaller increases for zanamivir and oseltamivir. A proline substitution (T146P) had a slightly lower (>700-fold) effect on the peramivir IC50 and also on the other NAIs. The presence of a second NA mutation at N169S combined with the T146P further increased the IC50 of peramivir (>7000-fold) and the other NAIs. A synergistic effect was also confirmed for dual NA mutations with G247D + I361V which showed a modest increase in the IC50 for oseltamivir (6-fold). Only one of two RG-viruses with the mutation G108E could be rescued and it had a high IC50 against zanamivir (>4000-fold) and laninamivir (>7000-fold), but a lower IC50 against oseltamivir (>200-fold). NA mutations H101L, A200T, D432G, H439P and H439R were also confirmed to somewhat reduce the in vitro susceptibility of influenza B viruses to the NAIs. Overall, this study identifies the potential impact of selected mutations on the clinical performance of NAIs when used to treat influenza B infection in humans.

Report on influenza viruses received and tested by the Melbourne WHO Collaborating Centre for Reference and Research on Influenza in 2019.

As part of its role in the World Health Organization's (WHO) Global Influenza Surveillance and Response System (GISRS), the WHO Collaborating Centre for Reference and Research on Influenza in Melbourne received a record total of 9,266 human influenza positive samples during 2019. Viruses were analysed for their antigenic, genetic and antiviral susceptibility properties. Selected viruses were propagated in qualified cells or embryonated hen's eggs for potential use in seasonal influenza virus vaccines. In 2019, influenza A(H3N2) viruses predominated over influenza A(H1N1)pdm09 and B viruses, accounting for a total of 51% of all viruses analysed. The majority of A(H1N1)pdm09, A(H3N2) and influenza B viruses analysed at the Centre were found to be antigenically similar to the respective WHO recommended vaccine strains for the Southern Hemisphere in 2019. However, phylogenetic analysis indicated that a significant proportion of circulating A(H3N2) viruses had undergone genetic drift relative to the WHO recommended vaccine strain for 2019. Of 5,301 samples tested for susceptibility to the neuraminidase inhibitors oseltamivir and zanamivir, four A(H1N1)pdm09 viruses showed highly reduced inhibition with oseltamivir, one A(H1N1)pdm09 virus showed highly reduced inhibition with zanamivir and three B/Victoria viruses showed highly reduced inhibition with zanamivir.

Detection of H275Y Mutation Conferring Oseltamivir Drug Resistance in Influenza A (H1N1) pdm09 Virus

In the treatment of influenza, Neuraminidase inhibitors (NAIs) (Oseltamivir and Zanamivir) play a major role. The emergence of variants of influenza A (H1N1) pdm09 virus resistant to Oseltamivir is a matter of great concern as it limits its usage. Therefore, vigilant monitoring for Oseltamivir-resistant viruses has been recommended by the World Health Organization (WHO). Our study aimed to screen the influenza A (H1N1) pdm09 virus for NAI drug resistance during the outbreak of 2015-16 in North-Western India. A total of 640 H1N1pdm09 virus-positive samples were screened for drug resistance to Oseltamivir by WHO allelic discrimination real-time RT-PCR protocol. The allelic discrimination PCR protocol can detect the presence of single nucleotide polymorphisms (SNPs), the H275Y mutation is detected by this method which causes resistance to Oseltamivir. Sanger sequencing of partial fragment of NA gene (fragment IV), of 90 samples were performed to confirm the presence of NA-H275Y mutation. Neuraminidase susceptibility of 20 randomly selected isolates to Oseltamivir was tested using NA inhibition chemiluminiscence based assay. Among 640 H1N1pdm09 positive samples tested, H275Y mutation was detected in one sample (0.15%) by PCR and confirmed by Sanger sequencing also. All the 20 isolates tested for NAI susceptibility by NA star assay were found to be sensitive to Oseltamivir. WHO allelic discrimination PCR is an easy, rapid and sensitive method for high-throughput detection of resistance to Oseltamivir. Systematic regular drug resistance surveillance of Influenza A is essential to monitor the emergence and spread of drug-resistant strains.

Sensitivity of influenza viruses to specific drugs in Russia during 2017−2020. The rare finds and perspective antivirals

BACKGROUND: The article presents the results of studying the effectiveness of anti-influenza drugs in Russia in the period 20172020. The sensitivity of circulating strains of influenza viruses A(H1N1)pdm09, A(H3N2) and B to neuraminidase inhibitors oseltamivir, zanamivir and rimantadine was determined. The analysis of scientific articles by both domestic and foreign researchers on new promising chemotherapy drugs for influenza viruses was carried out.
 AIMS: study of the sensitivity of influenza viruses circulating strains to specific anti-influenza drugs in the framework of monitoring in the period 20172020.
 MATERIALS AND METHODS: The study was conducted within the framework of epidemiological surveillance of the influenza viruses circulation using the collection of the influenza etiology and epidemiology laboratory with the following criteria for selecting strains isolated from pregnant women, patients with complicated flu infection and severe acute respiratory infection (SARI), lethal outcomes, as well as patients undergoing treatment with specific drugs. Virological, immunological, and molecular genetic methods were used in the study, and following drugs substances were used: oseltamivir carboxylate, zanamivir, and rimantadine.
 RESULTS: For the period 20172020, the sensitivity of 541 influenza A and B viruses epidemic strains to anti-influenza drugs was studied. Most of the studied strains remained sensitive to neuraminidase inhibitors. The exceptions were: in 2017/2018 5 strains of the influenza A(H1N1)pdm09 virus resistant to oseltamivir and 2 strains of the influenza B virus, one with reduced sensitivity to oseltamivir, the second to zanamivir; in 2019/2020 influenza A(H1N1)pdm09 virus strain with reduced sensitivity to both drugs.
 In the 2018/2019 season, an influenza A/Moscow/246/2018 A(H1N1)pdm09 strain was found to be sensitive to rimantadine.
 CONCLUSIONS: The main and most common genetic markers of influenza viruses resistance to specific drugs are: for oseltamivir substitution H274Y in the influenza A(H1N1)pdm09 virus NA; for rimantadine substitution S31N in the influenza A(H1N1)pdm09 and A(H3N2) viruses M2 protein. Taking into account the low frequency of strains with reduced sensitivity to drugs with antineuraminidase activity, can confidently approve that they remain the drugs of choice for the treatment and prevention of influenza infection.

Report on influenza viruses received and tested by the Melbourne WHO Collaborating Centre for Reference and Research on Influenza during 2020-2021.

As part of its role in the World Health Organization's (WHO) Global Influenza Surveillance and Response System (GISRS), the WHO Collaborating Centre for Reference and Research on Influenza in Melbourne received a total of 2,393 human influenza positive samples between 1 January 2020 and 31 December 2021 (2020: n = 2,021 samples; 2021: n = 372 samples). Viruses were analysed for their antigenic, genetic and antiviral susceptibility properties. Selected viruses were propagated in qualified cells or embryonated hen's eggs for potential use in seasonal influenza virus vaccines. During 2020-2021, influenza A viruses (A(H1N1)pdm09 in 2020 and A(H3N2) in 2021) predominated over influenza B viruses. In 2020, the majority of A(H1N1)pdm09, A(H3N2) and influenza B viruses analysed at the Centre were found to be antigenically similar to the respective WHO recommended vaccine strains for the southern hemisphere in 2020. In 2021, the majority of A(H1N1)pdm09 and A(H3N2) viruses were found to be antigenically distinct relative to the WHO recommended vaccine strains for the southern hemisphere in 2021. Of the influenza B viruses analysed at the Centre, 46.7% were found to be antigenically distinct to the respective WHO recommended vaccine strains. Of 1,538 samples tested for susceptibility to the neuraminidase inhibitors oseltamivir and zanamivir (in 2020, n = 1,374; in 2021, n = 164), two A(H1N1)pdm09 viruses showed highly reduced inhibition against oseltamivir, and one A(H1N1)pdm09 virus showed highly reduced inhibition against zanamivir. All of these samples were received in 2020.

Glycosylation in a Common Mechanism of Colitis and Sepsis

Glycosylation is a post‐translation modification common to proteins produced in the secretory pathway. In the bloodstream and at cell surfaces, glycan structures operate in multiple processes including the determination of protein half‐life, abundance, and function. Disruption of the normal mammalian N‐glycan repertoire can occur in part from acquired environmental and metabolic factors, and are responsible for the pathogenesis of multiple common diseases including autoimmunity, diabetes, colitis, and sepsis. Microbial infections are common throughout mammalian lifespan and can be life‐threatening as in the case of sepsis. Bacterial pathogens have also been considered as a possible environmental triggers precipitating chronic diseases including the human inflammatory bowel diseases (IBDs). For example, seasonal bacterial infections have been associated with increases in clinical diagnoses of IBD. To test the possibility that recurrent self‐resolving bacterial infections may trigger chronic disease, we developed a mouse model of recurrent human food poisoning using low‐titer non‐lethal gastric infections of Gram‐negative Salmonella enterica Typhimurium (ST). The ST pathogen produces a greater human disease burden than any other foodborne bacterial pathogen in the United States and is endemic throughout the world. Although the mouse rapidly cleared multiple recurrent low titer ST infections, colitis developed after several months, escalating in severity and failing to resolve after the cessation of infection. ST progressively disabled a mechanism of host protection by inducing intestinal neuraminidase activity with Neu3 gene induction among duodenal enterocytes. This accelerated the rate of molecular aging of intestinal alkaline phosphatase (IAP) measured by extent of de‐sialylation and led to its rapid endocytic clearance, thereby diminishing IAP levels in the small intestine and colon. Pathogenesis required host Tlr4 function through neuraminidase induction which was deleterious to the host as IAP deficiency increased the abundance of the LPS‐phosphate endotoxin in the colon. Oral administration of IAP or the marketed anti‐viral neuraminidase inhibitor Zanamivir were therapeutic reducing both LPS‐phosphate and inflammatory tissue damage. Humans with IBD have been reported with low IAP levels, while IAP‐null humans develop IBD. In both mouse and human colitis studies, IAP augmentation is remarkably therapeutic. This pathogenic mechanism emanating from altered protein glycosylation may be a trigger of one or more forms of IBD among the human population. In related studies where the ST pathogen gains access to the bloodstream in sepsis, we discovered a similar mechanism operating among Gram‐negative pathogens including ST and E. coli (EC). Both pathogens operated through Tlr4 to increase host neuraminidases Neu1 and Neu3 in the bloodstream, and thereby diminished alkaline phosphatase activities via endocytic clearance by the Ashwell‐Morell receptor of hepatocytes. These findings reveal a similar pathogenic process operating among ST and EC pathogens to subvert host protection by diminishing the expression of anti‐inflammatory enzymes. These findings further support published data and rationale for clinical trials augmenting alkaline phosphatase activity in sepsis and IBD patients.Support or Funding InformationNIH DK048247; HL131474; HL125352; AI141394

Report on influenza viruses received and tested by the Melbourne WHO Collaborating Centre for Reference and Research on Influenza in 2018

As part of its role in the World Health Organization's (WHO) Global Influenza Surveillance and Response System (GISRS), the WHO Collaborating Centre for Reference and Research on Influenza in Melbourne received a total of 3993 human influenza-positive samples during 2018. Viruses were analysed for their antigenic, genetic and antiviral susceptibility properties. Selected viruses were propagated in qualified cells or hens' eggs for use as potential seasonal influenza vaccine virus candidates. In 2018, influenza A(H1)pdm09 viruses predominated over influenza A(H3) and B viruses, accounting for a total of 53% of all viruses analysed. The majority of A(H1)pdm09, A(H3) and influenza B viruses analysed at the Centre were found to be antigenically similar to the respective WHO-recommended vaccine strains for the Southern Hemisphere in 2018. However, phylogenetic analysis indicated that a significant proportion of circulating A(H3) viruses had undergone genetic drift relative to the WHO-recommended vaccine strain for 2018. Of 2864 samples tested for susceptibility to the neuraminidase inhibitors oseltamivir and zanamivir, three A(H1)pdm09 viruses showed highly reduced inhibition by oseltamivir, while one B/Victoria virus showed highly reduced inhibition by both oseltamivir and zanamivir.

Specific Influenza Therapy: Current State and Prospects (Review)

Introduction. Respiratory infections are among the leaders in morbidity and mortality worldwide. The most severe cases of the disease are most often caused by the flu virus. Currently, there are many ways of specific prevention and treatment of influenza infection, but their effectiveness is far from ideal. This is due to the high variability of the influenza virus and the subsequent occurrence of resistance to the drugs used. In this regard, the improvement and development of antiviral drugs is an urgent task.Text. Influenza virus is an RNA-containing virus that causes massive epidemics and pandemics. Specific influenza prophylaxis includes vaccination. However, antigenic variability of the virus reduces the effectiveness of the vaccine, which requires constant costly development of its more advanced modifications. Specific treatment for influenza infection includes several classes of drugs, such as neuraminidase (NA) inhibitors oseltamivir, zanamivir and M2 protein inhibitors amantadine, rimantadine. At one time, these drugs were quite effective. But the formed resistance of influenza viruses to these drugs requires the creation of new or modifications of existing antiviral agents. Among the new domestic developments of antiviral drugs, histidyl-1-adamantainethylamine, which is a modification of the rimantadine molecule, has shown sufficient antiviral activity at the stage of preclinical studies. A representative of another class of drugs is arbidol (umifenovir), an inhibitor of hemagglutinin (HA) of the influenza virus. According to studies, the drug has high profiles of efficacy and safety, but the recommendation of the World Health Organization is to continue clinical trials. Currently, clinical studies of new classes of drugs are underway – baloxavir marboxil and favipiravir. Baloxavir marboxyl is a prodrug that is converted in vivo to baloxavir, an inhibitor of cap-dependent endonuclease. Favipiravir is an inhibitor of RNA-dependent RNA polymerase. In vitro studies in cell culture and in vivo in laboratory animals have shown higher efficacy of these drugs than the above with minimal toxicity.Conclusion. The rapid evolution of the influenza virus leads to a gradual decrease in the effectiveness of modern antiviral drugs. New compounds targeting targets important for virus reproduction are in clinical trials. The future of the fight against influenza depends on the outcome of these tests, according to which the compounds can become effective drugs for the prevention and treatment of influenza.

Diversity of A(H5N1) clade 2.3.2.1c avian influenza viruses with evidence of reassortment in Cambodia, 2014-2016.

In Cambodia, highly pathogenic avian influenza A(H5N1) subtype viruses circulate endemically causing poultry outbreaks and zoonotic human cases. To investigate the genomic diversity and development of endemicity of the predominantly circulating clade 2.3.2.1c A(H5N1) viruses, we characterised 68 AIVs detected in poultry, the environment and from a single human A(H5N1) case from January 2014 to December 2016. Full genomes were generated for 42 A(H5N1) viruses. Phylogenetic analysis shows that five clade 2.3.2.1c genotypes, designated KH1 to KH5, were circulating in Cambodia during this period. The genotypes arose through multiple reassortment events with the neuraminidase (NA) and internal genes belonging to H5N1 clade 2.3.2.1a, clade 2.3.2.1b or A(H9N2) lineages. Phylogenies suggest that the Cambodian AIVs were derived from viruses circulating between Cambodian and Vietnamese poultry. Molecular analyses show that these viruses contained the hemagglutinin (HA) gene substitutions D94N, S133A, S155N, T156A, T188I and K189R known to increase binding to the human-type α2,6-linked sialic acid receptors. Two A(H5N1) viruses displayed the M2 gene S31N or A30T substitutions indicative of adamantane resistance, however, susceptibility testing towards neuraminidase inhibitors (oseltamivir, zanamivir, lananmivir and peramivir) of a subset of thirty clade 2.3.2.1c viruses showed susceptibility to all four drugs. This study shows that A(H5N1) viruses continue to reassort with other A(H5N1) and A(H9N2) viruses that are endemic in the region, highlighting the risk of introduction and emergence of novel A(H5N1) genotypes in Cambodia.

Report on influenza viruses received and tested by the Melbourne WHO Collaborating Centre for Reference and Research on Influenza in 2017

As part of its role in the World Health Organization’s (WHO) Global Influenza Surveillance and Response System (GISRS), the WHO Collaborating Centre for Reference and Research on Influenza in Melbourne received a record total of 5866 human influenza positive samples during 2017. Viruses were analysed for their antigenic, genetic and antiviral susceptibility properties and were propagated in qualified cells and hens’ eggs for use as potential seasonal influenza vaccine virus candidates. In 2017, influenza A(H3) viruses predominated over influenza A(H1)pdm09 and B viruses, accounting for a total of 54% of all viruses analysed. The majority of A(H1)pdm09, A(H3) and influenza B viruses analysed at the Centre were found to be antigenically similar to the respective WHO recommended vaccine strains for the Southern Hemisphere in 2017. However, phylogenetic analysis indicated that the majority of circulating A(H3) viruses had undergone genetic drift relative to the WHO recommended vaccine strain for 2017. Of 3733 samples tested for susceptibility to the neuraminidase inhibitors oseltamivir and zanamivir, only two A(H1)pdm09 viruses and one A(H3) virus showed highly reduced inhibition by oseltamivir, while just one A(H1)pdm09 virus showed highly reduced inhibition by zanamivir.