Inactivated influenza virus vaccine is efficient and reduces IL‐4 and IL‐6 in allergic asthma mice

Background

Allergic asthma is a globally respiratory inflammatory disease. Influenza virus is a respiratory pathogen that causes yearly epidemics and results in high rates of morbidity and mortality. Patients with allergic asthma had a more severe symptom and a higher mortality when they were infected with influenza virus. Hence, influenza vaccination is recommended for patients with asthma.

Objectives

We evaluated the efficacy and effects of influenza vaccination on allergic asthma in a mouse model.

Methods

Ovalbumin‐immunized mice were inoculated with inactivated influenza virus A/Puerto Rico/8/34 (PR8) as vaccines and morbidity or mortality and allergic asthma features of these mice were analyzed.

Results

Mice inoculated with inactivated PR8 induced high levels of anti‐PR8 IgG2a and upregulation of Toll‐like receptor (TLR) 7. Vaccinated allergic mice were healthy when they were challenged with live influenza virus while none of non‐vaccinated allergic mice survived. Furthermore, inactivated influenza virus vaccine induced neither extra airway inflammation nor asthma features such as IgE, airway hyper‐reactivity, and eosinophilia in allergic mice. Particularly, decreased frequency of immune cell infiltrated airways and Th2 cytokines IL‐4 and IL‐6 production in the bronchoalveolar lavage fluid were noted in vaccinated allergic mice. These results suggested that inactivated influenza virus vaccine is efficient to protect allergic mice from further influenza infection, and it does not exacerbate but reduces IL‐4 and IL‐6 of allergic asthma.

Conclusion

Influenza vaccination is essential and efficient for allergic subjects to protect influenza virus infection.     

Infection of swine ex vivo tissues with avian viruses including H7N9 and correlation with glycomic analysis

Objectives

Swine have been regarded as intermediate hosts in the spread of influenza from birds to humans but studies of the sialylated glycans that comprise their respiratory tract have not been extensively studied in the past. This study analyzed the sialylated N‐glycan and O‐glycan profile of swine trachea and lung and correlated this with ex‐vivo infection of swine explants with avian influenza viruses.

Sample

Lungs and tracheal samples were obtained from normal farm and laboratory raised swine and used for ex vivo infection as well as mass spectrometric analysis. Infection of the ex vivo tissues used high pathogenic and low pathogenic avian viruses including the novel H7N9 virus that emerged in China in early 2013.

Main outcome measures

Assessment of successful replication was determined by TCID50 as well as virus immunohistochemistry. The N‐glycan and O‐glycan profiles were measured by MALDI‐TOF and sialylated linkages were determined by sialidase treatment. Lectin binding histochemistry was also performed on formalin fixed tissue samples with positive binding detected by chromogen staining.

Results

The swine respiratory tract glycans differed from the human respiratory tact glycans in two main areas. There was a greater abundance of Gal‐α‐Gal linkages resulting in a relative decrease in sialylated glycans. The swine respiratory tract also had a greater proportion of glycans containing Neu5Gc and Siaα2‐6 glycans than the human respiratory tract. Infection with avian viruses was confined primarily to lung bronchioles rather than trachea and parenchyma.

Conclusions

In contrast to previous studies we found that there was not as much expression of Siaα2‐3 glycans on the surface of the trachea. Infection of Siaα2‐3 binding avian viruses was restricted to the lower respiratory tract bronchioles. This finding may diminish the ability of the swine to act as an intermediary in the transmission of avian viruses to humans.     

A novel approach for preparation of the antisera reagent for potency determination of inactivated H7N9 influenza vaccines

Background

The potency of inactivated influenza vaccines is determined using a single‐radial immunodiffusion (SRID) assay and requires standardized reagents consisting of a Reference Antigen and an influenza strain‐specific antiserum. Timely availability of reagents is a critical step in influenza vaccine production, and the need for backup approaches for reagent preparation is an important component of pandemic preparedness.

Objectives

When novel H7N9 viruses emerged in China in 2013, candidate inactivated H7N9 influenza vaccines were developed for evaluation in clinical trials, and reagents were needed to measure vaccine potency.

Methods

We previously described an alternative approach for generating strain‐specific potency antisera, utilizing modified vaccinia virus Ankara vectors to produce influenza hemagglutinin (HA)‐containing virus‐like particles (VLPs) for immunization. Vector‐produced HA antigen is not dependent upon the success of the traditional bromelain‐digestion and HA purification.

Results

Antiserum for H7N9 vaccines, produced after immunization of sheep with preparations of bromelain‐HA (br‐HA), was not optimal for the SRID assay, and the supply of antiserum was limited. However, antiserum obtained from sheep boosted with VLPs containing H7 HA greatly improved the ring quality in the SRID assay. Importantly, this antiserum worked well with both egg‐ and cell‐derived antigen and was distributed to vaccine manufacturers.

Conclusions

Utilizing a previously developed approach for preparing vaccine potency antiserum, we have addressed a major bottleneck encountered in preparation of H7N9 vaccine reagents. The combination of br‐HA and mammalian VLPs for sequential immunization represents the first use of an alternative approach for producing an influenza vaccine potency antiserum.     

Transmission dynamics of pandemic influenza A(H1N1)pdm09 virus in humans and swine in backyard farms in Tumbes,Peru

Objectives

We aimed to determine the frequency of pH1N1 transmission between humans and swine on backyard farms in Tumbes, Peru.

Design

Two‐year serial cross‐sectional study comprising four sampling periods: March 2009 (pre‐pandemic), October 2009 (peak of the pandemic in Peru), April 2010 (1st post‐pandemic period), and October 2011 (2nd post‐pandemic period).

Sample

Backyard swine serum, tracheal swabs, and lung sample were collected during each sampling period.

Main outcome measures

We assessed current and past pH1N1 infection in swine through serological testing, virus culture, and RT‐PCR and compared the results with human incidence data from a population‐based active surveillance cohort study in Peru.

Results

Among 1303 swine sampled, the antibody prevalence to pH1N1 was 0% pre‐pandemic, 8% at the peak of the human pandemic (October 2009), and 24% in April 2010 and 1% in October 2011 (post‐pandemic sampling periods). Trends in swine seropositivity paralleled those seen in humans in Tumbes. The pH1N1 virus was isolated from three pigs during the peak of the pandemic. Phylogenetic analysis revealed that these viruses likely represent two separate human‐to‐swine transmission events in backyard farm settings.

Conclusions

Our findings suggest that human‐to‐swine pH1N1 transmission occurred during the pandemic among backyard farms in Peru, emphasizing the importance of interspecies transmission in backyard pig populations. Continued surveillance for influenza viruses in backyard farms is warranted.     

Heterosubtypic cross‐protection induced by whole inactivated influenza virus vaccine in mice: influence of the route of vaccine administration

Background

Development of influenza vaccines capable of inducing broad protection against different virus subtypes is necessary given the ever‐changing viral genetic landscape. Previously, we showed that vaccination with whole inactivated virus (WIV) induces heterosubtypic protection against lethal virus infection in mice. Whole inactivated virus‐induced cross‐protection was found to be mediated primarily by flu‐specific CD8+ T cells.

Objectives

As it has been demonstrated that the route of vaccine administration strongly influences both the quantity and quality of vaccine‐induced immunity, in this study, we determined which route of WIV administration induces optimal heterosubtypic cross‐protection.

Methods

We compared the magnitude of the immune response and heterosubtypic protection against lethal A/PR/8/34 (H1N1) infection after subcutaneous (SC), intramuscular (IM), and intranasal (IN) vaccination with A/NIBRG‐14 (H5N1) WIV.

Results

Subcutaneous and IM administration was superior to IN administration of influenza WIV in terms of flu‐specific CD8+ T‐cell induction and protection of mice against lethal heterosubtypic challenge. Surprisingly, despite the very low flu‐specific CD8+ T‐cell responses detected in IN‐vaccinated mice, these animals were partially protected, most likely due to cross‐reactive IgA antibodies.

Conclusion

The results of this study show that the magnitude of WIV‐induced flu‐specific CD8+ T‐cell activity depends on the applied vaccination route. We conclude that parenteral administration of WIV vaccine, in particular IM injection, is superior to IN vaccine delivery for the induction of heterosubtypic cross‐protection and generally appears to elicit stronger immune responses than mucosal vaccination with WIV.     

Oxygen free radical involvement in acute lung injury induced by H5N1 virus in mice

Background

Acute lung injury is an important cause of death in humans infected with H5N1. It has been found that oxygen free radicals (OFRs) are elevated in lung tissue during influenza virus infections. In this study, we used a mouse model to explore the role of OFRs in acute lung injury caused by H5N1 viral infection.

Methods

Four‐ to six‐week‐old male specific pathogen‐free BALB/c mice were inoculated intranasally with 10⁵ 50% tissue culture infective doses (TCID₅₀) of highly pathogenic A/Chicken/Hebei/108/2002 (H5N1) viruses and were then given 1000 IU of lauric acid modified superoxide dismutase (LA‐SOD) by intraperitoneal injection, starting 2 days post‐infection and continuing for 6 days.

Results

The extent of lung injury and the concentration of OFRs were higher, and the SOD activity was lower in H5N1 virus‐infected mice than that in uninfected control mice on days 3, 6, and 7 post‐inoculation. Weak amelioration of clinical signs, a minor decrease in the total mortality and the extent of lung injury, and the lower OFRs concentration were seen in the LA‐SOD treatment group, but a reduction in lung virus titers was not observed in the LA‐SOD treatment at all time points.

Conclusions

The LA‐SOD treatment has a mild inhibitory effect on H5N1 influenza virus infection in mice. OFRs, therefore, might play an important role in the pathogenesis of acute lung injury induced by H5N1 virus.     

Emergence of H3N2pM‐like and novel reassortant H3N1 swine viruses possessing segments derived from the A (H1N1)pdm09 influenza virus,Korea

Background

Human‐to‐swine transmission of the pandemic H1N1 2009 [A(H1N1)pdm09] virus in pig populations resulted in reassortment events with endemic swine influenza viruses worldwide.

Objective

We investigated whether A(H1N1)pdm09‐derived reassortant viruses are present in South Korea and sought to determine the pathogenic potential of the novel swine viruses.

Methods

Pig lung tissues were collected from commercially slaughtered pigs. Isolated swine influenza viruses were genetically analyzed and characterized in vitro and in vivo.

Results

We identified reassortant H3N2 (H3N2pM‐like) and H3N1 swine viruses containing A(H1N1)pdm09‐like segments in Korean pigs that are genetically closely related to strains recently detected in pigs and humans in North America. Although the H3N2pM‐like and novel H3N1 reassortants demonstrated efficient replication in mice and ferrets, all the H3N1 strains exhibited growth advantage over the representative H3N2pM‐like virus in human airway cells. Interestingly, A/swine/Korea/CY02‐07/2012(H3N1) and A/swine/Korea/CY03‐13/2012(H3N1) reassortants were more readily transmitted to respiratory‐droplet‐contact ferrets compared with the H3N2pM‐like (A/swine/Korea/CY02‐10/2012) isolate. Furthermore, serologic evaluation showed poor antigenicity to contemporary reference human seasonal H3N2 vaccine strains.

Conclusions

We report here for the first time the isolation of H3N2pM‐like viruses outside North America and of novel reassortant swine H3N1 viruses with A(H1N1)pdm09‐derived genes. Apart from further complicating the genetic diversity of influenza A viruses circulating in domestic pigs, our data also indicate that these strains could potentially pose threat to public health asserting the need for continuous virus monitoring in these ecologically important hosts.     

Improving the representativeness of influenza viruses shared within the WHO Global Influenza Surveillance and Response System

Background

Sharing influenza viruses within the WHO Global Influenza Surveillance and Response System is crucial for monitoring evolution of influenza viruses.

Objectives

Analysis of timeliness and geographic representativeness of viruses shared by National Influenza Centres (NICs) in the WHO European Region with the London WHO Collaborating Centre for Reference and Research on Influenza for the Northern Hemisphere''s 2010–2011 and 2011–2012 influenza seasons.

Materials and methods

Data from NICs on influenza‐positive specimens shared with WHO CC London for the above‐mentioned influenza seasons were analyzed for timeliness of sharing with respect to the February deadline (31 January) for inclusion in the WHO consultations on the composition of influenza virus vaccines for the Northern Hemisphere and geographic representativeness.

Results

The 2010–2011 and 2011–2012 seasons were different in terms of the seasonal pattern, the timing of the epidemic, and the dominant virus. Consistent patterns of virus sharing across the seasons were observed. Approximately half the viruses collected before the deadline were not shared within the deadline; the average delay between date of specimen collection and shipment receipt was 3 and 1·5 months for the first and second season, respectively.

Conclusion

A baseline was provided for future work on enhancement of specimen sharing in the WHO European Region and improving the vaccine virus selection process. Greater insight into virus selection criteria applied by countries and the causes of delays in shipment are needed to understand the representativeness of viruses shared and to assess the importance of this for vaccine strain selection.     

Epidemiological and virological findings during multiple outbreaks of equine influenza in South America in 2012

Background

In 2012, equine influenza (EI) virus was confirmed as the cause of outbreaks of respiratory disease in horses throughout South America. In Uruguay and Argentina, hundreds of vaccinated thoroughbred horses in training and racing facilities were clinically affected.

Objective

To characterise the EI viruses detected during the outbreak in Uruguay and Argentina.

Methods

Virus was detected in nasopharyngeal swabs by a pan‐reactive influenza type A real‐time RT‐PCR. The nucleotide sequence of the HA1 gene was determined and analysed phylogenetically using mega 5 software. Amino acid sequences alignments were constructed and virus was antigenically characterised with specific ferret antisera. Paired serum samples were tested by haemagglutination inhibition and single radial haemolysis.

Results

The diagnosis of EIV was confirmed by real‐time RT‐PCR, virus isolation and serological testing. The phylogenetic analysis of HA1 gene sequences of 18 EI viruses indicated that all of them belong to clade 1 of the Florida sublineage of the American lineage and are closely related to viruses isolated in the United States in 2012. The HA1 of viruses identified in horses in racing facilities in Maroñas, Uruguay, and in Palermo, Argentina, displayed 100% amino acid sequence identity and were identical to that of a virus isolated in Dubai in 2012, from vaccinated endurance horses recently imported from Uruguay.

Conclusions

The surveillance data reported illustrate the international spread of EI viruses and support the recommendations of the OIE expert surveillance panel to include viruses of the Florida sublineage in vaccines.     

Unusually high impact of influenza B during the early 2012–2013 influenza season in Wales – epidemiology and clinical analysis of the first 100 cases

Background

Influenza B is often regarded as the milder form of the disease. The early 2012–2013 season in Wales saw the highest rate of influenza B‐associated primary care consultations since 1994–1995 and considerable hospitalisations.

Objectives

This report summarises features of the first 100 confirmed cases during 2012–2013 in Wales.

Methods

Case information was sourced from routine laboratory testing and virological surveillance.

Results and conclusions

Influenza B (Yamagata lineage) viruses dominated, mainly affecting younger adults, admission to critical care was unexpectedly common. Low vaccine uptake amongst at‐risk patients may have contributed to the burden of influenza in secondary care in Wales.     

Effect of passive immunization on immunogenicity and protective efficacy of vaccination against a Mexican low‐pathogenic avian H5N2 influenza virus

Background

Despite the use of vaccines, low‐pathogenic (LP) H5N2 influenza viruses have continued to circulate and evolve in chickens in Mexico since 1993, giving rise to multiple genetic variants. Antigenic drift is partially responsible for the failure to control H5N2 influenza by vaccination; the contribution of maternal antibodies to this problem has received less attention.

Methods

We investigated the effect of different antisera on the efficacy of vaccination and whether booster doses of vaccine can impact immune suppression.

Results

While single doses of inactivated oil emulsion vaccine to currently circulating H5N2 influenza viruses provide partial protection from homologous challenge, chickens that receive high‐titer homologous antisera intraperitoneally before vaccination showed effects ranging from added protection to immunosuppression. Post‐infection antisera were less immunosuppressive than antisera obtained from field‐vaccinated chickens. Homologous, post‐infection chicken antisera provided initial protection from virus challenge, but reduced the induction of detectable antibody responses. Homologous antisera from field‐vaccinated chickens were markedly immunosuppressive, annulling the efficacy of the vaccine and leaving the chickens as susceptible to infection as non‐vaccinated birds. Booster doses of vaccine reduced the immunosuppressive effects of the administered sera.

Conclusion

Vaccine efficacy against LP H5N2 in Mexico can be severely reduced by maternal antibodies. Source‐dependent antisera effects offer the possibility of further elucidation of the immunosuppressive components involved.     

EV‐D68 infection in children with asthma exacerbation and pneumonia in Mexico City during 2014 autumn

Background

Human enterovirus D68 (EV‐D68) recently caused an increase in mild‐to‐severe pediatric respiratory cases in North America and some European countries. Even though few of these children presented with acute paralytic disease, direct causal relationship cannot yet be assumed.

Objectives

The purposes of this report were to describe the clinical findings of an outbreak of EV‐D68 infection in Mexico City and identify the genetic relationship with previously reported strains.

Patients/Methods

Between September and December 2014, 126 nasopharyngeal samples (NPS) of hospitalized children <15 years of age with ARI were tested for the presence of respiratory viruses using a multiplex RT‐qPCR and EV‐D68‐specific RT‐qPCR. Clinical, epidemiological, and demographic data were collected and associated with symptomatology and viral infections. Phylogenetic analyses were performed using VP1 region.

Results

Enterovirus/rhinovirus infection was detected in 40 patients (31·7%), of which 24 patients were EV‐D68‐positive. EV‐D68 infection prevailed over September and October 2014 and was associated with neutrophilia and lymphopenia, and patients were more likely to develop hypoxemia. Phylogenetic analyses showed that Mexican EV‐D68 belongs to the new B1 clade.

Conclusions

This is the first EV‐D68 outbreak described in Mexico and occurred few weeks after the United States reported similar infections. Although EV‐D68 belongs to new B1 clade, no neurological affection was observed.     

Serotype and genetic diversity of human rhinovirus strains that circulated in Kenya in 2008

Background

Human rhinoviruses (HRVs) are a well‐established cause of the common cold and recent studies indicated that they may be associated with severe acute respiratory illnesses (SARIs) like pneumonia, asthma, and bronchiolitis. Despite global studies on the genetic diversity of the virus, the serotype diversity of these viruses across diverse geographic regions in Kenya has not been characterized.

Objectives

This study sought to characterize the serotype diversity of HRV strains that circulated in Kenya in 2008.

Methods

A total of 517 archived nasopharyngeal samples collected in a previous respiratory virus surveillance program across Kenya in 2008 were selected. Participants enrolled were outpatients who presented with influenza‐like (ILI) symptoms. Real‐time RT‐PCR was employed for preliminary HRV detection. HRV‐positive samples were amplified using RT‐PCR and thereafter the nucleotide sequences of the amplicons were determined followed by phylogenetic analysis.

Results

Twenty‐five percent of the samples tested positive for HRV. Phylogenetic analysis revealed that the Kenyan HRVs clustered into three main species comprising HRV‐A (54%), HRV‐B (12%), and HRV‐C (35%). Overall, 20 different serotypes were identified. Intrastrain sequence homology among the Kenyan strains ranged from 58% to 100% at the nucleotide level and 55% to 100% at the amino acid level.

Conclusion

These results show that a wide range of HRV serotypes with different levels of nucleotide variation were present in Kenya. Furthermore, our data show that HRVs contributed substantially to influenza‐like illness in Kenya in 2008.     

Evaluation of twenty‐two rapid antigen detection tests in the diagnosis of Equine Influenza caused by viruses of H3N8 subtype

Background

Equine influenza (EI) is a highly contagious disease caused by viruses of the H3N8 subtype. The rapid diagnosis of EI is essential to reduce the disease spread. Many rapid antigen detection (RAD) tests for diagnosing human influenza are available, but their ability to diagnose EI has not been systematically evaluated.

Objectives

The aim of this study was to compare the performance of 22 RAD tests in the diagnosis of EI.

Methods

The 22 RAD tests were performed on fivefold serial dilutions of EI virus to determine their detection limits. The four most sensitive RAD tests (ImmunoAce Flu, BD Flu examan, Quick chaser Flu A, B and ESPLINE Influenza A&B‐N) were further evaluated using nasopharyngeal samples collected from experimentally infected and naturally infected horses. The results were compared to those obtained using molecular tests.

Results

The detection limits of the 22 RAD tests varied hugely. Even the four RAD tests showing the best sensitivity were 125‐fold less sensitive than the molecular techniques. The duration of virus detection in the experimentally infected horses was shorter using the RAD tests than using the molecular techniques. The RAD tests detected between 27% and 73% of real‐time RT‐PCR‐positive samples from naturally infected horses.

Conclusions

The study demonstrated the importance of choosing the right RAD tests as only three of 22 were fit for diagnosing EI. It was also indicated that even RAD tests with the highest sensitivity serve only as an adjunct to molecular tests because of the potential for false‐negative results.     

Cost‐effectiveness analysis of antiviral treatment in the management of seasonal influenza A: point‐of‐care rapid test versus clinical judgment

Background

A point‐of‐care rapid test (POCRT) may help early and targeted use of antiviral drugs for the management of influenza A infection.

Objective

(i) To determine whether antiviral treatment based on a POCRT for influenza A is cost‐effective and, (ii) to determine the thresholds of key test parameters (sensitivity, specificity and cost) at which a POCRT based‐strategy appears to be cost effective.

Methods

An hybrid « susceptible, infected, recovered (SIR) » compartmental transmission and Markov decision analytic model was used to simulate the cost‐effectiveness of antiviral treatment based on a POCRT for influenza A in the social perspective. Data input parameters used were retrieved from peer‐review published studies and government databases. The outcome considered was the incremental cost per life‐year saved for one seasonal influenza season.

Results

In the base‐case analysis, the antiviral treatment based on POCRT saves 2 lives/100 000 person‐years and costs $7600 less than the empirical antiviral treatment based on clinical judgment alone, which demonstrates that the POCRT‐based strategy is dominant. In one and two way‐sensitivity analyses, results were sensitive to the POCRT accuracy and cost, to the vaccination coverage as well as to the prevalence of influenza A. In probabilistic sensitivity analyses, the POCRT strategy is cost‐effective in 66% of cases, for a commonly accepted threshold of $50 000 per life‐year saved.

Conclusion

The influenza antiviral treatment based on POCRT could be cost‐effective in specific conditions of performance, price and disease prevalence.     

Macroeconomic impact of a mild influenza pandemic and associated policies in Thailand,South Africa and Uganda: a computable general equilibrium analysis

Background

Previous research has demonstrated the value of macroeconomic analysis of the impact of influenza pandemics. However, previous modelling applications focus on high‐income countries and there is a lack of evidence concerning the potential impact of an influenza pandemic on lower‐ and middle‐income countries.

Objectives

To estimate the macroeconomic impact of pandemic influenza in Thailand, South Africa and Uganda with particular reference to pandemic (H1N1) 2009.

Methods

A single‐country whole‐economy computable general equilibrium (CGE) model was set up for each of the three countries in question and used to estimate the economic impact of declines in labour attributable to morbidity, mortality and school closure.

Results

Overall GDP impacts were less than 1% of GDP for all countries and scenarios. Uganda''s losses were proportionally larger than those of Thailand and South Africa. Labour‐intensive sectors suffer the largest losses.

Conclusions

The economic cost of unavoidable absence in the event of an influenza pandemic could be proportionally larger for low‐income countries. The cost of mild pandemics, such as pandemic (H1N1) 2009, appears to be small, but could increase for more severe pandemics and/or pandemics with greater behavioural change and avoidable absence.