Integrated analysis of microRNA-mRNA expression in A549 cells infected with influenza A viruses (IAVs) from different host species

2019 ◽  
Vol 263 ◽  
pp. 34-46 ◽  
Author(s):  
Jie Gao ◽  
Lingxi Gao ◽  
Rui Li ◽  
Zhenping Lai ◽  
Zengfeng Zhang ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Host Species ◽  
Influenza A ◽  
A549 Cells ◽  
Integrated Analysis ◽  
Influenza A Viruses

scholarly journals Apoptotic and Early Innate Immune Responses to PB1-F2 Protein of Influenza A Viruses Belonging to Different Subtypes in Human Lung Epithelial A549 Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-12
Author(s):  
Gunisha Pasricha ◽  
Sanjay Mukherjee ◽  
Alok K. Chakrabarti
Keyword(s):  
Inflammatory Response ◽  
Influenza A ◽  
H5n1 Virus ◽  
A549 Cells ◽  
Terminal Region ◽  
Influenza Viruses ◽  
Influenza A Viruses ◽  
Highly Pathogenic ◽  
Lung Epithelial ◽  
Pathogenic H5n1

PB1-F2 is a multifunctional protein and contributes to the pathogenicity of influenza A viruses. PB1-F2 is known to have strain and cell specific functions. In this study we have investigated the apoptotic and inflammatory responses of PB1-F2 protein from influenza viruses of diverse pathogenicities in A549 lung epithelial cells. Overexpression of PB1-F2 resulted in apoptosis and heightened inflammatory response in A549 cells. Comparison revealed that the response varied with each subtype. PB1-F2 protein from highly pathogenic H5N1 virus induced least apoptosis but maximum inflammatory response. Results indicated that apoptosis was mediated through death receptor ligands TNFα and TRAIL via Caspase 8 activation. Significant induction of cytokines/chemokines CXCL10, CCL5, CCL2, IFNα, and IL-6 was noted in A549 cells transfected with PB1-F2 gene construct of 2008 West Bengal H5N1 virus (H5N1-WB). On the contrary, PB1-F2 construct from 2007 highly pathogenic H5N1 isolate (H5N1-M) with truncated N-terminal region did not evoke as exuberant inflammatory response as the other H5N1-WB with full length PB1-F2, signifying the importance of N-terminal region of PB1-F2. Sequence analysis revealed that PB1-F2 proteins derived from different influenza viruses varied at multiple amino acid positions. The secondary structure prediction showed each of the PB1-F2 proteins had distinct helix-loop-helix structure. Thus, our data substantiate the notion that the contribution of PB1-F2 to influenza pathogenicity is greatly strain specific and involves multiple host factors. This data demonstrates that PB1-F2 protein of influenza A virus, when expressed independently is minimally apoptotic and strongly influences the early host response in A549 cells.

scholarly journals Aptamer Profiling of A549 Cells Infected with Low-Pathogenicity and High-Pathogenicity Influenza Viruses

Viruses ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 1028 ◽  
Author(s):  
Kevin M. Coombs ◽  
Philippe F. Simon ◽  
Nigel J. McLeish ◽  
Ali Zahedi-Amiri ◽  
Darwyn Kobasa
Keyword(s):  
Influenza A ◽  
Human Lung ◽  
A549 Cells ◽  
Influenza Viruses ◽  
Post Translational Modification ◽  
Emerging Pathogens ◽  
Influenza A Viruses ◽  
Human Lung Cells ◽  
Wide Range ◽  
High Pathogenicity

Influenza A viruses (IAVs) are important animal and human emerging and re-emerging pathogens that are responsible for yearly seasonal epidemics and sporadic pandemics. IAVs cause a wide range of clinical illnesses, from relatively mild infections by seasonal strains, to acute respiratory distress during infections with highly pathogenic avian IAVs (HPAI). For this study, we infected A549 human lung cells with lab prototype A/PR/8/34 (H1N1) (PR8), a seasonal H1N1 (RV733), the 2009 pandemic H1N1 (pdm09), or with two avian strains, an H5N1 HPAI strain or an H7N9 strain that has low pathogenicity in birds but high pathogenicity in humans. We used a newly-developed aptamer-based multiplexed technique (SOMAscan®) to examine >1300 human lung cell proteins affected by the different IAV strains, and identified more than 500 significantly dysregulated cellular proteins. Our analyses indicated that the avian strains induced more profound changes in the A549 global proteome compared to all tested low-pathogenicity H1N1 strains. The PR8 strain induced a general activation, primarily by upregulating many immune molecules, the seasonal RV733 and pdm09 strains had minimal effect upon assayed molecules, and the avian strains induced significant downregulation, primarily in antimicrobial response, cardiovascular and post-translational modification systems.

scholarly journals 3-Indoleacetonitrile Is Highly Effective in Treating Influenza A Virus Infection In Vitro and In Vivo

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1433
Author(s):  
Xuejin Zhao ◽  
Lianzhong Zhao ◽  
Ya Zhao ◽  
Kun Huang ◽  
Wenxiao Gong ◽  
...  
Keyword(s):  
Influenza A ◽  
Mdck Cells ◽  
A549 Cells ◽  
H1n1 Influenza ◽  
Indole Derivatives ◽  
Influenza A Viruses ◽  
Toxic Concentration ◽  
Influenza Activity

Influenza A viruses are serious zoonotic pathogens that continuously cause pandemics in several animal hosts, including birds, pigs, and humans. Indole derivatives containing an indole core framework have been extensively studied and developed to prevent and/or treat viral infection. This study evaluated the anti-influenza activity of several indole derivatives, including 3-indoleacetonitrile, indole-3-carboxaldehyde, 3-carboxyindole, and gramine, in A549 and MDCK cells. Among these compounds, 3-indoleacetonitrile exerts profound antiviral activity against a broad spectrum of influenza A viruses, as tested in A549 cells. Importantly, in a mouse model, 3-indoleacetonitrile with a non-toxic concentration of 20 mg/kg effectively reduced the mortality and weight loss, diminished lung virus titers, and alleviated lung lesions of mice lethally challenged with A/duck/Hubei/WH18/2015 H5N6 and A/Puerto Rico/8/1934 H1N1 influenza A viruses. The antiviral properties enable the potential use of 3-indoleacetonitrile for the treatment of IAV infection.

scholarly journals Analysis of the Variability in the Non-Coding Regions of Influenza A Viruses

2018 ◽  
Vol 5 (3) ◽  
pp. 76 ◽  
Author(s):  
Jessica Benkaroun ◽  
Gregory Robertson ◽  
Hugh Whitney ◽  
Andrew Lang
Keyword(s):  
Influenza Virus ◽  
Host Species ◽  
Influenza A ◽  
Wild Bird ◽  
Coding Region ◽  
Influenza A Viruses ◽  
Protein Coding ◽  
Coding Regions ◽  
Negative Sense ◽  
Specific Sequences

The genomes of influenza A viruses (IAVs) comprise eight negative-sense single-stranded RNA segments. In addition to the protein-coding region, each segment possesses 5′ and 3′ non-coding regions (NCR) that are important for transcription, replication and packaging. The NCRs contain both conserved and segment-specific sequences, and the impacts of variability in the NCRs are not completely understood. Full NCRs have been determined from some viruses, but a detailed analysis of potential variability in these regions among viruses from different host groups and locations has not been performed. To evaluate the degree of conservation in NCRs among different viruses, we sequenced the NCRs of IAVs isolated from different wild bird host groups (ducks, gulls and seabirds). We then extended our study to include NCRs available from the National Center for Biotechnology Information (NCBI) Influenza Virus Database, which allowed us to analyze a wider variety of host species and more HA and NA subtypes. We found that the amount of variability within the NCRs varies among segments, with the greatest variation found in the HA and NA and the least in the M and NS segments. Overall, variability in NCR sequences was correlated with the coding region phylogeny, suggesting vertical coevolution of the (coding sequence) CDS and NCR regions.

scholarly journals Long-term culture of human lung adenocarcinoma A549 cells enhances the replication of human influenza A viruses

2019 ◽  
Vol 100 (10) ◽  
pp. 1345-1349 ◽  
Author(s):  
Michiko Ujie ◽  
Kosuke Takada ◽  
Maki Kiso ◽  
Yuko Sakai-Tagawa ◽  
Mutsumi Ito ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Influenza A ◽  
Human Lung ◽  
A549 Cells ◽  
Cell Types ◽  
Influenza Viruses ◽  
Human Influenza ◽  
Influenza A Viruses ◽  
Human Lung Adenocarcinoma

Long-term culture of the human lung adenocarcinoma cell line A549 promotes the differentiation of these cells toward an alveolar type II cell phenotype. Here, we evaluated the susceptibility of long-term cultured A549 cells to human influenza viruses. A549 cells were cultured continuously for 25 days (D25-A549) or 1 day (D1-A549) in Ham’s F12K medium. Six human influenza A viruses grew much faster in D25-A549 cells than in D1-A549 cells; however, two influenza B viruses replicated poorly in both cell types. Two avian influenza viruses replicated efficiently in both cell types, with similar titres. Expression levels of human virus receptors were higher in D25-A549 cells than in D1-A549 cells. D25-A549 cells thus more efficiently support the replication of human influenza A viruses compared with D1-A549 cells. Our data suggest that long-term cultured A549 cells will be useful for influenza A virus research.

scholarly journals 1231. VIR-2482: A potent and broadly neutralizing antibody for the prophylaxis of influenza A illness

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S635-S636
Author(s):  
Matteo Samuele Pizzuto ◽  
Fabrizia Zatta ◽  
Andrea Minola ◽  
Alessia Peter ◽  
Katja Culap ◽  
...  
Keyword(s):  
Half Life ◽  
Influenza A ◽  
Lethal Dose ◽  
A549 Cells ◽  
Neutralizing Antibody ◽  
Life Extension ◽  
Influenza A Viruses ◽  
Head Region ◽  
Lethal Challenge

Abstract Background Influenza A viruses are responsible for seasonal epidemics and represent a constant pandemic threat. Influenza vaccines induce predominantly antibodies against the head region of hemagglutinin (HA) and are strain specific. Vaccine effectiveness is often suboptimal due to mismatch with drifting viruses and an inadequate immune response. Broadly neutralizing monoclonal antibodies (mAbs) targeting the conserved stem-region of HA may provide protection through multiple seasons and cover strains with pandemic potential. We report pre-clinical data on VIR-2482, a fully human anti-HA stem mAb with half-life extending Fc mutations. Methods Binding of VIR-2482 to a panel of influenza HAs and neutralization of H1N1 and H3N2 viruses were measured by ELISA and microneutralization. Epitope conservation was evaluated using 49,462 HA sequences retrieved from GiSAID. Engagement of human FcγRs by VIR-2482 was assessed by biolayer interferometry. Antibody-dependendent cell-mediated cytoxicity (ADCC) was measured via in vitro killing of A549 cells expressing H1-HA glycoprotein by human NK cells. Complement-dependent cytotoxicity (CDC) was evaluated by incubating VIR-2482 with H1N1 infected cells in the presence of guinea pig complement. Protection studies were performed in Balb/c mice given VIR-2482 24h before intranasal infection with a lethal dose of H1N1 PR8 and H3N2 HK/68. Results VIR-2482 binds to the HA proteins representing all 18 influenza A HA subtypes and neutralizes a broad panel of H1N1 and H3N2 viruses spanning almost 100 years of evolution. Bioinformatic analysis revealed >98.8% conservation for the majority of key contact residues examined from sequences retrieved for H1N1 and H3N2 between 2009-2019. The half-life extending mutations in the Fc portion do not affect the ability of the antibody to engage FcγRIIIa, FcγRIIa, and C1q as evidenced by their lack of impact on ADCC and CDC in vitro. Prophylactic administration of VIR-2482 protects Balb/c mice from infection with lethal challenge doses of H1N1 and H3N2 viruses. Conclusion The attributes of potency, broad recognition of a highly conserved epitope, retention of high-level effector functions in addition to half-life extension support the development of VIR-2482 as a universal prophylactic for influenza A illness. Disclosures Matteo Samuele Pizzuto, PhD, VIR Biotechnology (Employee) Fabrizia Zatta, n/a, Vir Biotechnology (Employee) Andrea Minola, MS, Vir Biotechnology (Employee) Alessia Peter, n/a, Vir Biotechnology (Employee) Katja Culap, n/a, Vir Biotechnology (Employee) Leah Soriaga, PhD, Vir Biotechnology (Employee) Anna De Marco, n/a, Vir Biotechnology (Employee) Barbara Guarino, PhD, Vir Biotechnology (Employee) Nadia Passini, n/a, Vir Biotechnology (Employee) David K. Hong, MD, Vir Biotechnology (Employee) Fabio Benigni, PhD, Vir Biotechnology (Employee) Christy Hebner, PhD, Vir Biotechnology (Employee) Aurelio Bonavia, PhD, Vir Biotechnology (Employee) Davide Corti, PhD, Vir Biotechnology (Employee)

Poly-ADP Ribosyl Polymerase 1 (PARP1) Regulates Influenza A Virus Polymerase

2018 ◽  
Author(s):  
Liset Westera ◽  
Alisha M. Jennings ◽  
Jad Maamary ◽  
Martin Schwemmle ◽  
Adolfo García-Sastre ◽  
...  
Keyword(s):  
Influenza A ◽  
Mammalian Species ◽  
A549 Cells ◽  
Virus Titer ◽  
Fold Increase ◽  
Host Factors ◽  
Barriers To Entry ◽  
Influenza A Viruses ◽  
Rdrp Activity ◽  
Epidemic Disease

Influenza A viruses (IAV) are evolutionarily successful pathogens, capable of infecting a number of avian and mammalian species, and responsible for pandemic and seasonal epidemic disease in humans. To infect new species, IAV typically must overcome a number of species barriers to entry, replication, and egress, even while virus replication is counter-acted by antiviral host factors and innate immune mechanisms. A number of host factors have been found to regulate the replication of IAV by interacting with the viral RNA-dependent RNA polymerase (RdRP). The host factor PARP1, a poly-ADP ribosyl polymerase, was required for optimal functions of human, swine, and avian influenza RdRP in human 293T cells. In IAV infection, PARP1 was required for syntheses of viral mRNA and vRNA progeny, and for synthesis of viral nucleoprotein (NP) in human lung A549 cells. Intriguingly, pharmacological inhibition of PARP1 enzymatic activity (PARylation) by 4-amino-1,8-naphthalamide led to a 4-fold increase in RdRP activity, and a 2.3-fold increase in virus titer. Exogenous expression of the natural PARylation inhibitor PARG also enhanced RdRP activity. These data suggest a virus-host interaction dynamic where PARP1 itself is required, but cellular PARylation has a distinct suppressive modality on influenza RdRP function.

Viral Fitness Landscapes in Diverse Host Species Reveal Multiple Evolutionary Lines for the NS1 Gene of Influenza A Viruses

2019 ◽  
Author(s):  
Raquel Muñoz-Moreno ◽  
Carles Martínez-Romero ◽  
Daniel Blanco-Melo ◽  
Christian V. Forst ◽  
Raffael Nachbagauer ◽  
...  
Keyword(s):  
Host Species ◽  
Influenza A ◽  
Fitness Landscapes ◽  
Viral Fitness ◽  
Influenza A Viruses ◽  
Ns1 Gene

scholarly journals A30 Avian influenza viruses in wild birds: Virus evolution in a multi-host ecosystem

2019 ◽  
Vol 5 (Supplement_1) ◽  
Author(s):  
Divya Venkatesh ◽  
Marjolein J Poen ◽  
Theo M Bestebroer ◽  
Rachel D Scheuer ◽  
Oanh Vuong ◽  
...  
Keyword(s):  
Avian Influenza ◽  
Host Species ◽  
Influenza A ◽  
Phylogenetic Trees ◽  
Influenza Viruses ◽  
Influenza A Viruses ◽  
East African ◽  
Bird Populations ◽  
Under Sampling ◽  
The Republic

Abstract Wild ducks and gulls are the major reservoirs for avian influenza A viruses (AIVs). The mechanisms that drive AIV evolution are complex at sites where various duck and gull species from multiple flyways breed, winter, or stage. The Republic of Georgia is located at the intersection of three migratory flyways: the Central Asian Flyway, East Asian/East African Flyway, and Black Sea/Mediterranean Flyway. For six consecutive years (2010–6), we collected AIV samples from various duck and gull species that breed, migrate, and overwinter in Georgia. We found substantial subtype diversity of viruses that varied in prevalence from year to year. Low pathogenic (LP)AIV subtypes included H1N1, H2N3, H2N5, H2N7, H3N8, H4N2, H6N2, H7N3, H7N7, H9N1, H9N3, H10N4, H10N7, H11N1, H13N2, H13N6, H13N8, and H16N3, plus two H5N5 and H5N8 highly pathogenic (HP)AIVs belonging to clade 2.3.4.4. Whole-genome phylogenetic trees showed significant host species lineage restriction for nearly all gene segments and significant differences for LPAIVs among different host species in observed reassortment rates, as defined by quantification of phylogenetic incongruence, and in nucleotide diversity. Hemagglutinin clade 2.3.4.4 H5N8 viruses, circulated in Eurasia during 2014–5 did not reassort, but analysis after its subsequent dissemination during 2016–7 revealed reassortment in all gene segments except NP and NS. Some virus lineages appeared to be unrelated to AIVs in wild bird populations in other regions with maintenance of local AIV viruses in Georgia, whereas other lineages showed considerable genetic inter-relationship with viruses circulating in other parts of Eurasia and Africa, despite relative under-sampling in the area.

scholarly journals Avian Influenza A Viruses Reassort and Diversify Differently in Mallards and Mammals

Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 509
Author(s):  
Ketaki Ganti ◽  
Anish Bagga ◽  
Juliana DaSilva ◽  
Samuel S. Shepard ◽  
John R. Barnes ◽  
...  
Keyword(s):  
Linkage Disequilibrium ◽  
Host Species ◽  
Guinea Pigs ◽  
Influenza A ◽  
Model System ◽  
Mammalian Host ◽  
Viral Diversity ◽  
Influenza A Viruses ◽  
Longitudinal Patterns ◽  
Novel Host

Reassortment among co-infecting influenza A viruses (IAVs) is an important source of viral diversity and can facilitate expansion into novel host species. Indeed, reassortment played a key role in the evolution of the last three pandemic IAVs. Observed patterns of reassortment within a coinfected host are likely to be shaped by several factors, including viral load, the extent of viral mixing within the host and the stringency of selection. These factors in turn are expected to vary among the diverse host species that IAV infects. To investigate host differences in IAV reassortment, here we examined reassortment of two distinct avian IAVs within their natural host (mallards) and a mammalian model system (guinea pigs). Animals were co-inoculated with A/wildbird/California/187718-36/2008 (H3N8) and A/mallard/Colorado/P66F1-5/2008 (H4N6) viruses. Longitudinal samples were collected from the cloaca of mallards or the nasal tract of guinea pigs and viral genetic exchange was monitored by genotyping clonal isolates from these samples. Relative to those in guinea pigs, viral populations in mallards showed higher frequencies of reassortant genotypes and were characterized by higher genotype richness and diversity. In line with these observations, analysis of pairwise segment combinations revealed lower linkage disequilibrium in mallards as compared to guinea pigs. No clear longitudinal patterns in richness, diversity or linkage disequilibrium were present in either host. Our results reveal mallards to be a highly permissive host for IAV reassortment and suggest that reduced viral mixing limits avian IAV reassortment in a mammalian host.

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