Primary chicken tracheal cell culture system for the study of infection with avian respiratory viruses

Zaffuto, K. M.; Estévez, Carlos; Afonso, Claudio L.

doi:10.1080/03079450701774850

Cited by 25 publications

(22 citation statements)

References 30 publications

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“…Such receptor specificity can explain the increased infectivity of the H9N2 virus in CTE compared to CEF cells. Similar results on susceptibility of the primary tracheal epithelial chicken cells to infection with H7N3 and H6N1 influenza subtypes have been reported (Zaffuto et al 2008;Shen et al 2011) suggesting that the ciliated cells support the influenza virus growth. However, other host factors such as glycan topology, concentration of invading viruses, local density of receptors, lipid raft microdomains, co-receptors or SA-independent receptors, may also be important determinants of AIV infectivity (Zhang 2009).…”

Section: Discussionsupporting

confidence: 81%

Impact of chicken-origin cells on adaptation of a low pathogenic influenza virus

et al. 2012

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Understanding the growth dynamics of influenza viruses is an essential step in virus replication and cell-adaptation. The aim of this study was to elucidate the growth kinetic of a low pathogenic avian influenza H9N2 subtype in chicken embryo fibroblast (CEF) and chicken tracheal epithelial (CTE) cells during consecutive passages. An egg-adapted H9N2 virus was seeded into both cell culture systems. The amount of infectious virus released into the cell culture supernatants at interval times post-infection were titered and plaque assayed. The results as well as cell viability results indicate that the infectivity of the influenza virus was different among these primary cells. The egg-adapted H9N2 virus featured higher infectivity in CTE than in CEF cells. After serial passages and plaque purifications of the virus, a CTE cell-adapted strain was generated which carried amino acid substitutions within the HA stem region. The strain showed faster replication kinetics in cell culture resulting in an increase in virus titer. Overall, the present study provides the impact of cell type, multiplicity of infection, cellular protease roles in virus infectivity and finally molecular characterization during H9N2 virus adaptation procedure.

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Section: Discussionsupporting

confidence: 81%

Impact of chicken-origin cells on adaptation of a low pathogenic influenza virus

et al. 2012

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“…Lee et al (13) have been suggested that H9N2 viruses belonging to G1 sub-lineage, may be better adapted to the human host and replicates efficiently in human alveolar epithelial cell line (A549) than other H9N2 sub-lineages. Due to the viral tropism, avian influenza viruses reach high titers when grown in chicken-origin cells (14)(15)(16). The efficient replication and infectivity of low pathogenic influenza viruses is achieved in the presence of supplemental trypsin or the type II transmembrane serine proteases (TMPRSS) (14,17).…”

Section: Introductionmentioning

confidence: 99%

Replication Efficiency of Influenza A Virus H9N2: A Comparative Analysis Between Different Origin Cell Types

Lebas

Shahsavandi

Mohammadi

et al. 2013

Jundishapur J Microbiol

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Background: Efficient isolation and detection of low pathogenic avian influenza viruses from surveillance samples continues to be a high priority. Currently, the new cell lines are considered for supporting the replication to high virus strains titers. Objectives: The replication efficiency of a low pathogenic avian influenza virus in different origin cells was evaluated under different conditions. Materials and Methods: Chicken embryo fibroblast (CEF) cell and human alveolar epithelial cell line (A549) were infected with H9N2 at a multiplicity of infection of 0.1. The amount of infectious virus released into the cell culture supernatants at various post-infection time intervals were tittered by tissue culture infectious dose (TCID50) assay. The impact of these cells adaptation was investigated by determination the virus genes nucleotide sequences. Results: The influenza virus infectivity was not significant difference in these cells in the presence of trypsin. The results of fusion assay and determination of cellular protease confirmed that A549 cells support virus entry with or without supplemental trypsin. However, the H9N2 virus showed lower titer and infectivity in the trypsin-free infected A549 cells within longer time. The comparative sequence analysis indicated several simultaneously nucleotide substitutions were occurred in NA of the virus replicated in A549 cells resulted in two fixed amino acid changes at positions G320 to A and G414 to A up to the fifth passage. Conclusions: After seven consecutive passages of both cell cultures, the H9N2 virus showed similar antigenicity, also no change on viral titer level and virus replication behavior in adaptation was found. The results highlighted the use of A549 cells for efficient virus isolation.

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“…In vitro cell culture can permit study of infection mechanisms under highly controlled conditions, although in vivo systems remain essential for studying responses to disease [23]. Given the demonstration by Petkov et al [24] that a Bu-1-F + and IgM + subpopulation in the bursa is targeted in IBDV infections, the potential of the present agglomerate system for study of NDV infection was examined.…”

Section: Discussionmentioning

confidence: 99%

Establishment of an In Vitro System Representing the Chicken Gut-Associated Lymphoid Tissue

et al. 2012

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The bursa of Fabricius is critical for B cell development and differentiation in chick embryos. This study describes the production in vitro, from dissociated cell suspensions, of cellular agglomerates with functional similarities to the chicken bursa. Co-cultivation of epithelial and lymphoid cells obtained from embryos at the appropriate developmental stage regularly led to agglomerate formation within 48 hours. These agglomerates resembled bursal tissue in having lymphoid clusters overlaid by well organized epithelium. Whereas lymphocytes within agglomerates were predominantly Bu-1a+, a majority of those emigrating onto the supporting membrane were Bu-1a− and IgM+. Both agglomerates and emigrant cells expressed activation-induced deaminase with levels increasing after 24 hours. Emigrating cells were actively proliferating at a rate in excess of both the starting cell population and the population of cells remaining in agglomerates. The potential usefulness of this system for investigating the response of bursal tissue to avian Newcastle disease virus (strain AF2240) was examined.

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Primary chicken tracheal cell culture system for the study of infection with avian respiratory viruses

Cited by 25 publications

References 30 publications

Impact of chicken-origin cells on adaptation of a low pathogenic influenza virus

Impact of chicken-origin cells on adaptation of a low pathogenic influenza virus

Replication Efficiency of Influenza A Virus H9N2: A Comparative Analysis Between Different Origin Cell Types

Establishment of an In Vitro System Representing the Chicken Gut-Associated Lymphoid Tissue

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