Design of peptide epitope from the neuraminidase protein of influenza A and influenza B towards short peptide vaccine development

Abstract Background A country-wide lab-based surveillance system for ILI and Severe Acute Respiratory Illness (SARI) with weekly sampling and reporting was established in 2008.This system was necessary for early detection of emerging novel influenza subtypes and timely response for influenza prevention and control. Objectives To assess the trends of Influenza-like-Illness(ILI) and to monitor the predominant circulating strains of influenza viruses through Lab based sentinel surveillance. Methods A cross-sectional study was conducted based on ten years (2007-2017) influenza surveillance data obtained from National Influenza Central Laboratory Pakistan (NICLP) from January to March 2018.Study was done from the data records and samples of suspected ILI patients and SARI patients received from all seven sentinel sites. An ILI case was defined as sudden onset of fever of ≥ 38 C° and cough, with onset within last 10 days, while patients with sudden onset of fever (>38 °C), cough/sore throat requiring hospital admission within 7 days were termed as SARI. Samples were tested at NICLP for confirmation of virus, typing and subtyping by RT-PCR. Results A total of 15885 samples were analyzed during ten years period, out of which 3475(21.9%) were found positive for influenza virus. Among positive samples 26(0.75%) were Influenza-A (H1N1), 550(38%) were A/H3N1,550(15.9%) were A/H3N1,1587(45.7%) were A/H1N1 pdm09and 1312(37.8%) were influenza B. Males were predominant(54%).Influenza Maximum cases were reported from age group 01->12 years(66%).Virus circulation was detected throughout the year along with few cases of seasonal A/H1N1 virus during late winter(January February) and spring(March). Influenza A/H3N2 virus circulation was mainly observed during summer months (August-October). Conclusions The findings of this study emphasize the need for continuous and comprehensive influenza surveillance to predict seasonal trends for vaccine development and to further fortify pandemic preparedness. Key messages The need for continuous and comprehensive influenza surveillance. Public health importance by pandemic preparedness.

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Balanced Cellular and Humoral Immune Responses Targeting Multiple Antigens in Adults Receiving a Quadrivalent Inactivated Influenza Vaccine

10.20944/preprints202104.0072.v1 ◽

2021 ◽

Author(s):

Esther Dawen Yu ◽

Alba Grifoni ◽

Aaron Sutherland ◽

Hannah Voic ◽

Eric Wang ◽

...

Keyword(s):

T Cell ◽

Immune Responses ◽

Influenza Vaccine ◽

Influenza A ◽

Mononuclear Cells ◽

Vaccine Development ◽

Influenza B ◽

Cell Reactivity ◽

Cell Immunity ◽

Influenza A H1n1

The role of T cell immunity has been acknowledged in recent vaccine development and evaluation. We tested the humoral and cellular immune responses to Flucelvax®, a quadrivalent inactivated seasonal influenza vaccine containing two influenza A (H1N1 Singapore/GP1908/2015 IVR-180 and H3N2 North Carolina/04/2016) and two influenza B (Iowa/06/2017 and Singapore/INFTT-16-0610/2016) virus strains, using peripheral blood mononuclear cells stimulated by pools of peptides overlapping all the individual influenza viral protein components. Baseline reactivity was detected against all four strains both at the level of CD4 and CD8 responses and targeting different proteins. CD4 T cell reactivity was mostly directed to HA/NA proteins in influenza B strains, and NP/M1/M2/NS1/NEP proteins in the case of the Influenza A strains. CD8 responses to both influenza A and B viruses preferentially targeted the more conserved core viral proteins. Following vaccination, both CD4 and CD8 responses against the various influenza antigens were increased in day 15 to day 91 post vaccination period and maintained a Th1 polarized profile. Importantly, no vaccine interference was detected, with the increased responses balanced across all 4 included viral strains for both CD4 and CD8 T cells, and targeting HA and multiple additional viral antigens.

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Balanced Cellular and Humoral Immune Responses Targeting Multiple Antigens in Adults Receiving a Quadrivalent Inactivated Influenza Vaccine

Vaccines ◽

10.3390/vaccines9050426 ◽

2021 ◽

Vol 9 (5) ◽

pp. 426

Author(s):

Esther Dawen Yu ◽

Alba Grifoni ◽

Aaron Sutherland ◽

Hannah Voic ◽

Eric Wang ◽

...

Keyword(s):

T Cell ◽

Immune Responses ◽

Influenza Vaccine ◽

Influenza A ◽

Mononuclear Cells ◽

Vaccine Development ◽

Influenza B ◽

Cell Reactivity ◽

Cell Immunity ◽

Influenza A H1n1

The role of T cell immunity has been acknowledged in recent vaccine development and evaluation. We tested the humoral and cellular immune responses to Flucelvax®, a quadrivalent inactivated seasonal influenza vaccine containing two influenza A (H1N1 Singapore/GP1908/2015 IVR-180 and H3N2 North Carolina/04/2016) and two influenza B (Iowa/06/2017 and Singapore/INFTT-16-0610/2016) virus strains, using peripheral blood mononuclear cells stimulated by pools of peptides overlapping all the individual influenza viral protein components. Baseline reactivity was detected against all four strains both at the level of CD4 and CD8 responses and targeting different proteins. CD4 T cell reactivity was mostly directed to HA/NA proteins in influenza B strains, and NP/M1/M2/NS1/NEP proteins in the case of the Influenza A strains. CD8 responses to both influenza A and B viruses preferentially targeted the more conserved core viral proteins. Following vaccination, both CD4 and CD8 responses against the various influenza antigens were increased in day 15 to day 91 post vaccination period, and maintained a Th1 polarized profile. Importantly, no vaccine interference was detected, with the increased responses balanced across all four included viral strains for both CD4 and CD8 T cells, and targeting HA and multiple additional viral antigens.

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Epidemiological characteristics of four common respiratory viral infections in children

Virology Journal ◽

10.1186/s12985-020-01475-y ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Guohong Zhu ◽

Dan Xu ◽

Yuanyuan Zhang ◽

Tianlin Wang ◽

Lingyan Zhang ◽

...

Keyword(s):

Respiratory Tract ◽

Viral Infection ◽

Influenza A ◽

Multiple Infection ◽

Influenza B ◽

Preschool Period ◽

Significant Difference ◽

Different Seasons ◽

Tract Infections ◽

Epidemiological Characteristics

Abstract Background Viruses are the main infectious agents of acute respiratory infections in children. We aim to describe the epidemiological characteristics of viral pathogens of acute respiratory tract infections in outpatient children. Methods From April 2018 to March 2019, the results of viral detection using oral pharyngeal swabs from 103,210 children with acute respiratory tract infection in the outpatient department of the Children’s Hospital, Zhejiang University School of Medicine, were retrospectively analyzed. Viral antigens, including adenovirus (ADV), influenza A (FLUA), influenza B (FLUB) and respiratory syncytial virus (RSV), were detected by the colloidal gold method. Results At least one virus was detected in 38,355 cases; the positivity rate was 37.2%. A total of 1910 cases of mixed infection with two or more viruses were detected, and the positivity rate of multiple infection was 1.9%. The ADV positivity rate was highest in the 3–6-year-old group (18.7%), the FLUA positivity rate was highest in the > 6-year-old group (21.6%), the FLUB positivity rate was highest in the > 6-year-old group (6.6%), and the RSV positivity rate was highest in the < 1-year-old group (10.6%). There was a significant difference in the positivity rate of viral infection among different age groups (χ2 = 1280.7, P < 0.001). The rate of positive viral infection was highest in winter (47.1%). The ADV infection rate was highest in spring (18.2%). The rates of FLUA and FLUB positivity were highest in winter (28.8% and 3.6%, respectively). The rate of RSV positivity was highest in autumn (17.4%). The rate of positive viral infection in different seasons was significantly different (χ2 = 6459.1, P < 0.001). Conclusions Viral infection rates in children differ for different ages and seasons. The positivity rate of ADV is highest in the preschool period and that of RSV is highest in infants; that of FLU increases with age. The total positive rate of viral infection in different seasons is highest in winter, as is the rate of FLU positivity.

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An evaluation of the InDevR FluChip-8G insight microarray assay in characterizing influenza a viruses

Tropical Diseases Travel Medicine and Vaccines ◽

10.1186/s40794-021-00133-7 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Emily S. Bailey ◽

Xinye Wang ◽

Mai-juan Ma ◽

Guo-lin Wang ◽

Gregory C. Gray

Keyword(s):

Influenza A ◽

Influenza Viruses ◽

Time Range ◽

Influenza B ◽

Influenza A Viruses ◽

Laboratory Methods ◽

Duke University ◽

Multiple Viral Strains ◽

Rapid Characterization

AbstractInfluenza viruses are an important cause of disease in both humans and animals, and their detection and characterization can take weeks. In this study, we sought to compare classical virology techniques with a new rapid microarray method for the detection and characterization of a very diverse, panel of animal, environmental, and human clinical or field specimens that were molecularly positive for influenza A alone (n = 111), influenza B alone (n = 3), both viruses (n = 13), or influenza negative (n = 2) viruses. All influenza virus positive samples in this study were first subtyped by traditional laboratory methods, and later evaluated using the FluChip-8G Insight Assay (InDevR Inc. Boulder, CO) in laboratories at Duke University (USA) or at Duke Kunshan University (China). The FluChip-8G Insight multiplexed assay agreed with classical virologic techniques 59 (54.1%) of 109 influenza A-positive, 3 (100%) of the 3 influenza B-positive, 0 (0%) of 10 both influenza A- and B-positive samples, 75% of 24 environmental samples including those positive for H1, H3, H7, H9, N1, and N9 strains, and 80% of 22 avian influenza samples. It had difficulty with avian N6 types and swine H3 and N2 influenza specimens. The FluChip-8G Insight assay performed well with most human, environmental, and animal samples, but had some difficulty with samples containing multiple viral strains and with specific animal influenza strains. As classical virology methods are often iterative and can take weeks, the FluChip-8G Insight Assay rapid results (time range 8 to 12 h) offers considerable time savings. As the FluChip-8G analysis algorithm is expected to improve over time with addition of new subtypes and sample matrices, the FluChip-8G Insight Assay has considerable promise for rapid characterization of novel influenza viruses affecting humans or animals.

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Differential Expression of Serum Exosome microRNAs and Cytokines in Influenza A and B Patients Collected in the 2016 and 2017 Influenza Seasons

Pathogens ◽

10.3390/pathogens10020149 ◽

2021 ◽

Vol 10 (2) ◽

pp. 149

Author(s):

Sreekumar Othumpangat ◽

William G. Lindsley ◽

Donald H. Beezhold ◽

Michael L. Kashon ◽

Carmen N. Burrell ◽

...

Keyword(s):

Healthy Volunteers ◽

Influenza A ◽

Influenza Infection ◽

Cell Types ◽

Airway Epithelial Cells ◽

Differentially Expressed ◽

Influenza B ◽

Airway Epithelial ◽

Monocyte Chemoattractant Protein 1 ◽

Novel Approach

MicroRNAs (miRNAs) have remarkable stability and are key regulators of mRNA transcripts for several essential proteins required for the survival of cells and replication of the virus. Exosomes are thought to play an essential role in intercellular communications by transporting proteins and miRNAs, making them ideal in the search for biomarkers. Evidence suggests that miRNAs are involved in the regulation of influenza virus replication in many cell types. During the 2016 and 2017 influenza season, we collected blood samples from 54 patients infected with influenza and from 30 healthy volunteers to identify the potential role of circulating serum miRNAs and cytokines in influenza infection. Data comparing the exosomal miRNAs in patients with influenza B to healthy volunteers showed 76 miRNAs that were differentially expressed (p < 0.05). In contrast, 26 miRNAs were differentially expressed between patients with influenza A (p < 0.05) and the controls. Of these miRNAs, 11 were commonly expressed in both the influenza A and B patients. Interferon (IFN)-inducing protein 10 (IP-10), which is involved in IFN synthesis during influenza infection, showed the highest level of expression in both influenza A and B patients. Influenza A patients showed increased expression of IFNα, GM-CSF, interleukin (IL)-13, IL-17A, IL-1β, IL-6 and TNFα, while influenza B induced increased levels of EGF, G-CSF, IL-1α, MIP-1α, and TNF-β. In addition, hsa-miR-326, hsa-miR-15b-5p, hsa-miR-885, hsa-miR-122-5p, hsa-miR-133a-3p, and hsa-miR-150-5p showed high correlations to IL-6, IL-15, IL-17A, IL-1β, and monocyte chemoattractant protein-1 (MCP-1) with both strains of influenza. Next-generation sequencing studies of H1N1-infected human lung small airway epithelial cells also showed similar pattern of expression of miR-375-5p, miR-143-3p, 199a-3p, and miR-199a-5p compared to influenza A patients. In summary, this study provides insights into the miRNA profiling in both influenza A and B virus in circulation and a novel approach to identify the early infections through a combination of cytokines and miRNA expression.

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Influenza: An Emerging and Re-emerging Public Health Threat in Nepal

Annals of Clinical Chemistry and Laboratory Medicine ◽

10.3126/acclm.v3i2.20737 ◽

2018 ◽

Vol 3 (2) ◽

pp. 1-2

Author(s):

Bishnu Prasad Upadhyay

Keyword(s):

Influenza Virus ◽

Influenza A ◽

Winter Season ◽

Emerging Infectious Diseases ◽

Influenza Viruses ◽

Influenza B ◽

Influenza A Viruses ◽

Influenza A H1n1 ◽

New Variant ◽

Seasonal Epidemics

Influenza virus type A and B are responsible for seasonal epidemics as well as pandemics in human. Influenza A viruses are further divided into two major groups namely, low pathogenic seasonal influenza (A/H1N1, A/H1N1 pdm09, A/H3N2) and highly pathogenic influenza virus (H5N1, H5N6, H7N9) on the basis of two surface antigens: hemagglutinin (HA) and neuraminidase (NA). Mutations, including substitutions, deletions, and insertions, are one of the most important mechanisms for producing new variant of influenza viruses. During the last 30 years; more than 50 viral threat has been evolved in South-East Asian countriesof them influenza is one of the major emerging and re-emerging infectious diseases of global concern. Similar to tropical and sub-tropical countries of Southeast Asia; circulation of A/H1N1 pdm09, A/H3N2 and influenza B has been circulating throughout the year with the peak during July-November in Nepal. However; the rate of infection transmission reach peak during the post-rain and winter season of Nepal.

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Multiplex loop-mediated isothermal amplification (M-LAMP) assay for the detection of influenza A/H1, A/H3 and influenza B can provide a specimen-to-result diagnosis in 40min with single genome copy sensitivity

Journal of Clinical Virology ◽

10.1016/j.jcv.2013.06.006 ◽

2013 ◽

Vol 58 (1) ◽

pp. 127-131 ◽

Cited By ~ 40

Author(s):

James Mahony ◽

Sylvia Chong ◽

David Bulir ◽

Alexandra Ruyter ◽

Ken Mwawasi ◽

...

Keyword(s):

Influenza A ◽

Lamp Assay ◽

Isothermal Amplification ◽

Influenza B ◽

Genome Copy ◽

Loop Mediated Isothermal Amplification ◽

Single Genome

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QUALITATIVE DIFFERENCES IN THE ANTIGENIC COMPOSITION OF INFLUENZA A VIRUS STRAINS

Journal of Experimental Medicine ◽

10.1084/jem.79.6.633 ◽

1944 ◽

Vol 79 (6) ◽

pp. 633-647 ◽

Cited By ~ 18

Author(s):

William F. Friedewald

Keyword(s):

Influenza A Virus ◽

Influenza A ◽

Serum Antibody ◽

Allantoic Fluid ◽

Influenza B Virus ◽

Influenza B ◽

Red Cell ◽

Homologous Virus ◽

Cell Adsorption ◽

Virus Strains

A study of the PR8, Christie, Talmey, W.S., and swine strains of influenza A virus by means of antibody absorption tests revealed the following findings: 1. Serum antibody could be specifically absorbed with allantoic fluid containing influenza virus or, more effectively, with concentrated suspensions of virus obtained from allantoic fluid by high-speed centrifugation or by the red cell adsorption and elution technique. Normal allantoic fluid, or the centrifugalized sediment therefrom, failed to absorb antibodies. Influenza B virus (Lee) caused no detectable absorption of antibody from antisera directed against influenza A virus strains, but it specifically absorbed antibody from Lee antisera. 2. The neutralizing, agglutination-inhibiting, and complement-fixing anti-bodies in ferret antisera were completely absorbed only by the homologous virus strain, even though 2 absorptions were carried out with large amounts of heterologous virus strains. 3. PR8 virus appeared to have the broadest range of specific antigenic components for it completely absorbed the heterologous antibodies in Christie and W.S. antisera and left only those antibodies which reacted with the respective homologous strains. The other virus strains (Christie, Talmey, W.S., swine) were more specific in the absorption of heterologous antibodies and completely removed only those antibodies which reacted with the absorbing virus. 4. The absorption tests revealed a higher degree of specificity and individuality of the virus strains than the various cross reactions previously reported. The strain specificity of PR8 virus was equally manifest in absorption tests with ferret sera and with human sera following vaccination. 5. The amount of homologous antibody remaining in a PR8 ferret serum after absorption with PR8 virus, obtained by the red cell adsorption and elution method, varied inversely as the concentration of virus used for absorption. A given concentration of virus, however, absorbed a greater percentage of neutralizing antibodies than either agglutination-inhibiting or complement-fixing antibodies.

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Community-acquired bacteraemia in COVID-19 in comparison to influenza A and influenza B: a retrospective cohort study

BMC Infectious Diseases ◽

10.1186/s12879-021-05902-5 ◽

2021 ◽

Vol 21 (1) ◽

Cited By ~ 1

Author(s):

Julinha M. Thelen ◽

A. G. ( Noud) Buenen ◽

Marjan van Apeldoorn ◽

Heiman F. Wertheim ◽

Mirjam H. A. Hermans ◽

...

Keyword(s):

Cohort Study ◽

Blood Culture ◽

The Netherlands ◽

Retrospective Cohort Study ◽

Retrospective Cohort ◽

Influenza A ◽

Blood Cultures ◽

Influenza B ◽

Significant Difference ◽

All Cause Mortality

Abstract Background During the coronavirus disease 2019 (COVID-19) pandemic in the Netherlands it was noticed that very few blood cultures from COVID-19 patients turned positive with clinically relevant bacteria. This was particularly evident in comparison to the number of positive blood cultures during previous seasonal epidemics of influenza. This observation raised questions about the occurrence and causative microorganisms of bacteraemia in COVID-19 patients, especially in the perspective of the widely reported overuse of antibiotics and the rising rate of antibiotic resistance. Methods We conducted a retrospective cohort study on blood culture results in influenza A, influenza B and COVID-19 patients presenting to two hospitals in the Netherlands. Our main outcome consisted of the percentage of positive blood cultures. The percentage of clinically relevant blood cultures, isolated bacteria and 30-day all-cause mortality served as our secondary outcomes. Results A total of 1331 viral episodes were analysed in 1324 patients. There was no statistically significant difference (p = 0.47) in overall occurrence of blood culture positivity in COVID-19 patients (9.0, 95% CI 6.8–11.1) in comparison to influenza A (11.4, 95% CI 7.9–14.8) and influenza B patients (10.4, 95% CI 7.1–13.7,). After correcting for the high rate of contamination, the occurrence of clinically relevant bacteraemia in COVID-19 patients amounted to 1.0% (95% CI 0.3–1.8), which was statistically significantly lower (p = 0.04) compared to influenza A patients (4.0, 95% CI 1.9–6.1) and influenza B patients (3.0, 95% CI 1.2–4.9). The most frequently identified bacterial isolates in COVID-19 patients were Escherichia coli (n = 2) and Streptococcus pneumoniae (n = 2). The overall 30-day all-cause mortality for COVID-19 patients was 28.3% (95% CI 24.9–31.7), which was statistically significantly higher (p = <.001) when compared to patients with influenza A (7.1, 95% CI 4.3–9.9) and patients with influenza B (6.4, 95% CI 3.8–9.1). Conclusions We report a very low occurrence of community-acquired bacteraemia amongst COVID-19 patients in comparison to influenza patients. These results reinforce current clinical guidelines on antibiotic management in COVID-19, which only advise utilization of antibiotics when a bacterial co-infection is suspected.

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