Temperature sensitivity on growth and/or replication of H1N1, H1N2 and H3N2 influenza A viruses isolated from pigs and birds in mammalian cells

Massin, Pascale; Simon, Gaëlle; Barbezange, Cyril; Deblanc, Céline; Oger, Aurélie; Marquet-Blouin, Estelle; Bougeard, Stéphanie; Werf, Sylvie van der; Jestin, Véronique

doi:10.1016/j.vetmic.2009.10.012

Cited by 38 publications

(43 citation statements)

References 44 publications

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“…SL3-10(C)/(¡) and SL5-3B(C)/(¡) were present in the 5 0 and 3 0 ends of the coding regions for M1 and M2, respectively, where packaging signals have been inferred to be located. 18,20 Since the body temperatures at which IAVs replicate vary between »33 C and »40 C among host species, 43 the stability of a particular vRNA structure may vary in different hosts. 30,45,46 However, stem base-pairings in SL3-10(C)/(¡) and SL5-3(C)/(¡) are retained in >99% of 1,884 sequences, and in silico analysis predicts these stemloop structures are stable under different temperature settings (37 C and 40 C) (data not shown), suggesting that they are common and conserved structures within the M segment among IAVs, and may play a role in productive IAV infection even in avian hosts whose body temperature is higher than in mammals.…”

Section: Discussionmentioning

confidence: 99%

Computational and molecular analysis of conserved influenza A virus RNA secondary structures involved in infectious virion production

et al. 2016

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As well as encoding viral proteins, genomes of RNA viruses harbor secondary and tertiary RNA structures that have been associated with functions essential for successful replication and propagation. Here, we identified stem-loop structures that are extremely conserved among 1,884 M segment sequences of influenza A virus (IAV) strains from various subtypes and host species using computational and evolutionary methods. These structures were predicted within the 3 0 and 5 0 ends of the coding regions of M1 and M2, respectively, where packaging signals have been previously proposed to exist. These signals are thought to be required for the incorporation of a single copy of 8 different negative-strand RNA segments (vRNAs) into an IAV particle. To directly test the functionality of conserved stem-loop structures, we undertook reverse genetic experiments to introduce synonymous mutations designed to disrupt secondary structures predicted at 3 locations and found them to attenuate infectivity of recombinant virus. In one mutant, predicted to disrupt stem loop structure at nucleotide positions 219-240, attenuation was more evident at increased temperature and was accompanied by an increase in the production of defective virus particles. Our results suggest that the conserved secondary structures predicted in the M segment are involved in the production of infectious viral particles during IAV replication.

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Section: Discussionmentioning

confidence: 99%

Computational and molecular analysis of conserved influenza A virus RNA secondary structures involved in infectious virion production

et al. 2016

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“…Many studies have described host-pathogen interactions in respiratory epithelial cell cultures derived from humans (Vos et al 2005) and from a variety of animals (Wang et al 2014), including mice (Parikh et al 2008), horses (Abraham et al 2011), cattle (Su et al 2013), and others. Airway epithelial cells have been widely used as a cell model in studies of the pathogenesis of human, murine, or swine respiratory pathogen infections (Massin et al 2010). For example, airway epithelial cells play a pivotal role in the pathological mechanisms of human influenza (Massin et al 2010) and asthma (Hahn 2002).…”

Section: Introductionmentioning

confidence: 99%

“…Airway epithelial cells have been widely used as a cell model in studies of the pathogenesis of human, murine, or swine respiratory pathogen infections (Massin et al 2010). For example, airway epithelial cells play a pivotal role in the pathological mechanisms of human influenza (Massin et al 2010) and asthma (Hahn 2002). In addition, in standard animal models of respiratory diseases, including asthma models in mice, rats, guinea pigs, and rabbits, airway epithelial cells have been considered to be pathological and therapeutic targets of airway diseases (Abraham et al 2011).…”

Section: Introductionmentioning

confidence: 99%

Establishment and characterization of a telomerase-immortalized canine bronchiolar epithelial cell line

Xie

Pang

Shan

et al. 2015

Appl Microbiol Biotechnol

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Dogs are susceptible to infectious diseases that occur primarily in the respiratory tract. The airway epithelium acts as a first line of defense and is constantly exposed to microorganisms present in the environment. Respiratory epithelial cells have recently gained wide use as a cell model for studying the pathogenesis of human, murine or swine respiratory pathogen infections. However, studies of the pathogenic mechanisms of canine pathogens have been hindered by the lack of reliable respiratory cell lines. Here, we cultured primary canine bronchiolar epithelial cells (CBECs), whose characteristics were confirmed by their expression of the epithelial cell-specific marker cytokeratin 18, and have provided protocols for their isolation and ex vivo expansion. Further, we established immortalized CBECs containing the human telomerase reverse transcriptase (hTERT) gene via transfection of primary CBECs with the recombinant plasmid pEGFP-hTERT. Immortalized bronchiolar epithelial cells (hTERT-CBECs) retain the morphological and functional features of primary CBECs, as indicated by reverse transcriptase polymerase chain reaction, proliferation assays, karyotype analysis, telomerase activity assay, and Western blotting, which demonstrate that hTERT-CBECs have higher telomerase activity, an extended proliferative lifespan, and a diploid complement of chromosomes, even after Passage 50. Moreover, this cell line is not transformed, as evaluated using soft agar assays and tumorigenicity analysis in nude mice, and can therefore be safely used in future studies. The isolation and establishment of stable hTERT-CBECs is of great importance for use as an in vitro model for mechanistic studies of canine pathogenic infections.

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“…In its more severe form, highly pathogenic (HP) avian IAVs cause systemic disease with high mortality and spread to numerous organs of the respiratory, digestive, and ner-vous systems (8,11,12). A difference in core body temperature between humans and avian species is a known factor in limiting interspecies transmission, as avian IAVs that have adapted to replication at 41°C demonstrate decreased polymerase activity at temperatures of 33 to 37°C, typical of the human respiratory tract (13)(14)(15)(16). However, the most important restriction for crossing the species barrier lies at the receptor level.…”

mentioning

confidence: 99%

Oncolytic Activity of Avian Influenza Virus in Human Pancreatic Ductal Adenocarcinoma Cell Lines

Kasloff

Pizzuto

Silic-Benussi

et al. 2014

J Virol

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Pancreatic ductal adenocarcinoma (PDA) is the most lethal form of human cancer, with dismal survival rates due to late-stage diagnoses and a lack of efficacious therapies. Building on the observation that avian influenza A viruses (IAVs) have a tropism for the pancreas in vivo, the present study was aimed at testing the efficacy of IAVs as oncolytic agents for killing human PDA cell lines. Receptor characterization confirmed that human PDA cell lines express the alpha-2,3-and the alpha-2,6-linked glycan receptor for avian and human IAVs, respectively. PDA cell lines were sensitive to infection by human and avian IAV isolates, which is consistent with this finding. Growth kinetic experiments showed preferential virus replication in PDA cells over that in a nontransformed pancreatic ductal cell line. Finally, at early time points posttreatment, infection with IAVs caused higher levels of apoptosis in PDA cells than gemcitabine and cisplatin, which are the cornerstone of current therapies for PDA. In the BxPC-3 PDA cell line, apoptosis resulted from the engagement of the intrinsic mitochondrial pathway. Importantly, IAVs did not induce apoptosis in nontransformed pancreatic ductal HPDE6 cells. Using a model based on the growth of a PDA cell line as a xenograft in SCID mice, we also show that a slightly pathogenic avian IAV significantly inhibited tumor growth following intratumoral injection. Taken together, these results are the first to suggest that IAVs may hold promise as future agents of oncolytic virotherapy against pancreatic ductal adenocarcinomas. IMPORTANCEDespite intensive studies aimed at designing new therapeutic approaches, PDA still retains the most dismal prognosis among human cancers. In the present study, we provide the first evidence indicating that avian IAVs of low pathogenicity display a tropism for human PDA cells, resulting in viral RNA replication and a potent induction of apoptosis in vitro and antitumor effects in vivo. These results suggest that slightly pathogenic IAVs may prove to be effective for oncolytic virotherapy of PDA and provide grounds for further studies to develop specific and targeted viruses, with the aim of testing their efficacy in clinical contexts.

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Temperature sensitivity on growth and/or replication of H1N1, H1N2 and H3N2 influenza A viruses isolated from pigs and birds in mammalian cells

Cited by 38 publications

References 44 publications

Computational and molecular analysis of conserved influenza A virus RNA secondary structures involved in infectious virion production

Computational and molecular analysis of conserved influenza A virus RNA secondary structures involved in infectious virion production

Establishment and characterization of a telomerase-immortalized canine bronchiolar epithelial cell line

Oncolytic Activity of Avian Influenza Virus in Human Pancreatic Ductal Adenocarcinoma Cell Lines

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