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  1. Article: Replication of influenza A viruses in an avian macrophage cell line.

    Lyon, J A / Hinshaw, V S

    The Journal of general virology

    1991  Volume 72 Pt 8, Page(s) 2011–2013

    Abstract: ... of a wide variety of influenza viruses and that this continuous avian cell line may prove useful ... macrophage cell line, HD11, to Ty/Ont. We found that Ty/Ont replicated in the HD11 cells to high titres ... unique to Ty/Ont, we also examined the replication of influenza viruses representative of all 13 HA ...

    Abstract The virulent avian influenza virus A/Ty/Ont/7732/66 (H5N9) (Ty/Ont) causes severe destruction of the lymphoid cells in infected birds. Previous studies have suggested that viral infection of macrophages may be involved. However, Ty/Ont failed to replicate productively in primary cultures of chicken macrophages. Therefore, in an effort to develop an in vitro system for our studies, we examined the susceptibility of an avian macrophage cell line, HD11, to Ty/Ont. We found that Ty/Ont replicated in the HD11 cells to high titres, as measured by haemagglutination (HA) assays and infectivity yields. To determine whether this property was unique to Ty/Ont, we also examined the replication of influenza viruses representative of all 13 HA subtypes and an attenuated variant of Ty/Ont. All of the tested viruses replicated in HD11 cells; the avirulent strains required the presence of trypsin in the culture medium whereas virulent viruses and the attenuated variant of Ty/Ont did not. These results suggest that the HD11 cells can support the replication of a wide variety of influenza viruses and that this continuous avian cell line may prove useful for in vitro studies on these viruses.
    MeSH term(s) Animals ; Cell Line ; Chickens ; Influenza A virus/physiology ; Macrophages/microbiology ; Virus Replication
    Language English
    Publishing date 1991-08
    Publishing country England
    Document type Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, U.S. Gov't, P.H.S.
    ZDB-ID 219316-4
    ISSN 1465-2099 ; 0022-1317
    ISSN (online) 1465-2099
    ISSN 0022-1317
    DOI 10.1099/0022-1317-72-8-2011
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  2. Article: Replication of influenza A viruses in an avian macrophage cell line

    Lyon, J.A / Hinshaw, V.S

    Journal of general virology. Aug 1991. v. 72 (pt.8)

    1991  

    Abstract: ... of a wide variety of influenza viruses and that this continuous avian cell line may prove useful ... macrophage cell line, HD11, to Ty/Ont. We found that Ty/Ont replicated in the HD11 cells to high titres ... unique to Ty/Ont, we also examined the replication of influenza viruses representative of all 13 HA ...

    Abstract The virulent avian influenza virus A/Ty/Ont/7732/66 (H5N9) (Ty/Ont) causes severe destruction of the lymphoid cells in infected birds. Previous studies have suggested that viral infection of macrophages may be involved. However, Ty/Ont failed to replicate productively in primary cultures of chicken macrophages. Therefore, in an effort to develop an in vitro system for our studies, we examined the susceptibility of an avian macrophage cell line, HD11, to Ty/Ont. We found that Ty/Ont replicated in the HD11 cells to high titres, as measured by haemagglutination (HA) assays and infectivity yields. To determine whether this property was unique to Ty/Ont, we also examined the replication of influenza viruses representative of all 13 HA subtypes and an attenuated variant of Ty/Ont. All of the tested viruses replicated in HD11 cells; the avirulent strains required the presence of trypsin in the culture medium whereas virulent viruses and the attenuated variant of Ty/Ont did not. These results suggest that the HD11 cells can support the replication of a wide variety of influenza viruses and that this continuous avian cell line may prove useful for in vitro studies on these viruses.
    Keywords poultry ; Influenza A virus ; macrophages ; cell culture
    Language English
    Dates of publication 1991-08
    Size p. 2011-2013.
    Document type Article
    ZDB-ID 219316-4
    ISSN 1465-2099 ; 0022-1317
    ISSN (online) 1465-2099
    ISSN 0022-1317
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: The hemagglutinin protein of highly pathogenic H5N1 influenza viruses overcomes an early block in the replication cycle to promote productive replication in macrophages.

    Cline, Troy D / Karlsson, Erik A / Seufzer, Bradley J / Schultz-Cherry, Stacey

    Journal of virology

    2012  Volume 87, Issue 3, Page(s) 1411–1419

    Abstract: ... in macrophages. However, studies investigating the productive replication of influenza viruses in macrophages ... However, they may also contribute to severe disease caused by the highly pathogenic avian (HPAI) H5N1 influenza viruses ... as the only subtype of influenza virus capable of productive replication in macrophages and establishes ...

    Abstract Macrophages are known to be one of the first lines of defense against influenza virus infection. However, they may also contribute to severe disease caused by the highly pathogenic avian (HPAI) H5N1 influenza viruses. One reason for this may be the ability of certain influenza virus strains to productively replicate in macrophages. However, studies investigating the productive replication of influenza viruses in macrophages have been contradictory, and the results may depend on both the type of macrophages used and the specific viral strain. In this work, we investigated the ability of H1 to H16 viruses to productively replicate in primary murine alveolar macrophages and RAW264.7 macrophages. We show that only a subset of HPAI H5N1 viruses, those that cause high morbidity and mortality in mammals, can productively replicate in macrophages, as measured by the release of newly synthesized virus particles into the cell supernatant. Mechanistically, we found that these H5 strains can overcome a block early in the viral life cycle leading to efficient nuclear entry, viral transcription, translation, and ultimately replication. Studies with reassortant viruses demonstrated that expression of the hemagglutinin gene from an H5N1 virus rescued replication of H1N1 influenza virus in macrophages. This study is the first to characterize H5N1 influenza viruses as the only subtype of influenza virus capable of productive replication in macrophages and establishes the viral gene that is required for this characteristic. The ability to productively replicate in macrophages is unique to H5N1 influenza viruses and may contribute to their increased pathogenesis.
    MeSH term(s) Animals ; Cell Line ; Hemagglutinin Glycoproteins, Influenza Virus/metabolism ; Influenza A Virus, H1N1 Subtype/genetics ; Influenza A Virus, H1N1 Subtype/pathogenicity ; Influenza A Virus, H5N1 Subtype/pathogenicity ; Influenza A Virus, H5N1 Subtype/physiology ; Macrophages/immunology ; Macrophages/virology ; Mice ; Mice, Inbred C57BL ; Reassortant Viruses/genetics ; Reassortant Viruses/pathogenicity ; Virulence Factors/metabolism ; Virus Replication
    Chemical Substances Hemagglutinin Glycoproteins, Influenza Virus ; Virulence Factors
    Language English
    Publishing date 2012-11-14
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.02682-12
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Tropism, replication competence, and innate immune responses of the coronavirus SARS-CoV-2 in human respiratory tract and conjunctiva: an analysis in ex-vivo and in-vitro cultures.

    Hui, Kenrie P Y / Cheung, Man-Chun / Perera, Ranawaka A P M / Ng, Ka-Chun / Bui, Christine H T / Ho, John C W / Ng, Mandy M T / Kuok, Denise I T / Shih, Kendrick C / Tsao, Sai-Wah / Poon, Leo L M / Peiris, Malik / Nicholls, John M / Chan, Michael C W

    The Lancet. Respiratory medicine

    2020  Volume 8, Issue 7, Page(s) 687–695

    Abstract: ... epithelial cells and macrophages. In-vitro studies included the highly pathogenic avian influenza H5N1 virus (H5N1 ... and innate immune responses in comparison with other coronavirus and influenza virus to provide ... syndrome-associated coronavirus (MERS-CoV), and 2009 pandemic influenza H1N1 (H1N1pdm) in ex-vivo cultures ...

    Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019, causing a respiratory disease (coronavirus disease 2019, COVID-19) of varying severity in Wuhan, China, and subsequently leading to a pandemic. The transmissibility and pathogenesis of SARS-CoV-2 remain poorly understood. We evaluate its tissue and cellular tropism in human respiratory tract, conjunctiva, and innate immune responses in comparison with other coronavirus and influenza virus to provide insights into COVID-19 pathogenesis.
    Methods: We isolated SARS-CoV-2 from a patient with confirmed COVID-19, and compared virus tropism and replication competence with SARS-CoV, Middle East respiratory syndrome-associated coronavirus (MERS-CoV), and 2009 pandemic influenza H1N1 (H1N1pdm) in ex-vivo cultures of human bronchus (n=5) and lung (n=4). We assessed extrapulmonary infection using ex-vivo cultures of human conjunctiva (n=3) and in-vitro cultures of human colorectal adenocarcinoma cell lines. Innate immune responses and angiotensin-converting enzyme 2 expression were investigated in human alveolar epithelial cells and macrophages. In-vitro studies included the highly pathogenic avian influenza H5N1 virus (H5N1) and mock-infected cells as controls.
    Findings: SARS-CoV-2 infected ciliated, mucus-secreting, and club cells of bronchial epithelium, type 1 pneumocytes in the lung, and the conjunctival mucosa. In the bronchus, SARS-CoV-2 replication competence was similar to MERS-CoV, and higher than SARS-CoV, but lower than H1N1pdm. In the lung, SARS-CoV-2 replication was similar to SARS-CoV and H1N1pdm, but was lower than MERS-CoV. In conjunctiva, SARS-CoV-2 replication was greater than SARS-CoV. SARS-CoV-2 was a less potent inducer of proinflammatory cytokines than H5N1, H1N1pdm, or MERS-CoV.
    Interpretation: The conjunctival epithelium and conducting airways appear to be potential portals of infection for SARS-CoV-2. Both SARS-CoV and SARS-CoV-2 replicated similarly in the alveolar epithelium; SARS-CoV-2 replicated more extensively in the bronchus than SARS-CoV. These findings provide important insights into the transmissibility and pathogenesis of SARS-CoV-2 infection and differences with other respiratory pathogens.
    Funding: US National Institute of Allergy and Infectious Diseases, University Grants Committee of Hong Kong Special Administrative Region, China; Health and Medical Research Fund, Food and Health Bureau, Government of Hong Kong Special Administrative Region, China.
    MeSH term(s) Adult ; Aged ; Aged, 80 and over ; Betacoronavirus/immunology ; Betacoronavirus/physiology ; COVID-19 ; Conjunctiva/immunology ; Conjunctiva/physiopathology ; Conjunctiva/virology ; Coronavirus Infections/immunology ; Coronavirus Infections/physiopathology ; Female ; Humans ; Immunity, Innate/immunology ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral/immunology ; Pneumonia, Viral/physiopathology ; Respiratory Mucosa/immunology ; Respiratory Mucosa/physiopathology ; Respiratory Mucosa/virology ; Respiratory System/immunology ; Respiratory System/physiopathology ; Respiratory System/virology ; SARS-CoV-2 ; Viral Tropism/physiology ; Virus Replication/physiology
    Keywords covid19
    Language English
    Publishing date 2020-05-07
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2686754-0
    ISSN 2213-2619 ; 2213-2600
    ISSN (online) 2213-2619
    ISSN 2213-2600
    DOI 10.1016/S2213-2600(20)30193-4
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  5. Article ; Online: Swine alveolar macrophage cell model allows optimal replication of influenza A viruses regardless of their origin.

    Kasloff, Samantha B / Weingartl, Hana M

    Virology

    2016  Volume 490, Page(s) 91–98

    Abstract: ... swine pulmonary alveolar macrophage cell line (IPAM 3D4/31) to determine whether this system would allow ... demonstrated over the last century. Eleven influenza A viruses from avian, human and swine hosts were evaluated ... represent a natural host cell model for influenza virus replication where the only condition requiring ...

    Abstract The importance of pigs in interspecies transmission of influenza A viruses has been repeatedly demonstrated over the last century. Eleven influenza A viruses from avian, human and swine hosts were evaluated for replication phenotypes at three physiologically relevant temperatures (41°C, 37°C, 33°C) in an immortalized swine pulmonary alveolar macrophage cell line (IPAM 3D4/31) to determine whether this system would allow for their efficient replication. All isolates replicated well in IPAMs at 37°C while clear distinctions were observed at 41°C and 33°C, correlating to species of origin of the PB2, reflected in distinct amino acid residue profiles rather than in one particular PB2 residue. A strong TNF-α response was induced by some mammalian but not avian IAVs, while other selected cytokines remained below detection levels. Porcine IPAMs represent a natural host cell model for influenza virus replication where the only condition requiring modification for optimal IAV replication, regardless of virus origin.
    MeSH term(s) Animals ; Birds ; Humans ; Influenza A virus/genetics ; Influenza A virus/isolation & purification ; Influenza A virus/physiology ; Influenza in Birds/immunology ; Influenza in Birds/virology ; Influenza, Human/virology ; Macrophages, Alveolar/immunology ; Macrophages, Alveolar/virology ; Orthomyxoviridae Infections/immunology ; Orthomyxoviridae Infections/veterinary ; Orthomyxoviridae Infections/virology ; Swine ; Swine Diseases/immunology ; Swine Diseases/virology ; Tumor Necrosis Factor-alpha/immunology ; Virus Cultivation ; Virus Replication
    Chemical Substances Tumor Necrosis Factor-alpha
    Language English
    Publishing date 2016-03
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 200425-2
    ISSN 1096-0341 ; 0042-6822
    ISSN (online) 1096-0341
    ISSN 0042-6822
    DOI 10.1016/j.virol.2016.01.006
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  6. Article ; Online: Widespread Virus Replication in Alveoli Drives Acute Respiratory Distress Syndrome in Aerosolized H5N1 Influenza Infection of Macaques.

    Wonderlich, Elizabeth R / Swan, Zachary D / Bissel, Stephanie J / Hartman, Amy L / Carney, Jonathan P / O'Malley, Katherine J / Obadan, Adebimpe O / Santos, Jefferson / Walker, Reagan / Sturgeon, Timothy J / Frye, Lonnie J / Maiello, Pauline / Scanga, Charles A / Bowling, Jennifer D / Bouwer, Anthea L / Duangkhae, Parichat A / Wiley, Clayton A / Flynn, JoAnne L / Wang, Jieru /
    Cole, Kelly S / Perez, Daniel R / Reed, Douglas S / Barratt-Boyes, Simon M

    Journal of immunology (Baltimore, Md. : 1950)

    2017  Volume 198, Issue 4, Page(s) 1616–1626

    Abstract: ... therapeutic approaches to prevent and treat human disease caused by highly pathogenic avian influenza viruses. ... Human infections with highly pathogenic avian influenza A (H5N1) virus are frequently fatal ... where target cells reside. We show that inhalation of aerosolized H5N1 influenza virus in cynomolgus macaques ...

    Abstract Human infections with highly pathogenic avian influenza A (H5N1) virus are frequently fatal but the mechanisms of disease remain ill-defined. H5N1 infection is associated with intense production of proinflammatory cytokines, but whether this cytokine storm is the main cause of fatality or is a consequence of extensive virus replication that itself drives disease remains controversial. Conventional intratracheal inoculation of a liquid suspension of H5N1 influenza virus in nonhuman primates likely results in efficient clearance of virus within the upper respiratory tract and rarely produces severe disease. We reasoned that small particle aerosols of virus would penetrate the lower respiratory tract and blanket alveoli where target cells reside. We show that inhalation of aerosolized H5N1 influenza virus in cynomolgus macaques results in fulminant pneumonia that rapidly progresses to acute respiratory distress syndrome with a fatal outcome reminiscent of human disease. Molecular imaging revealed intense lung inflammation coincident with massive increases in proinflammatory proteins and IFN-α in distal airways. Aerosolized H5N1 exposure decimated alveolar macrophages, which were widely infected and caused marked influx of interstitial macrophages and neutrophils. Extensive infection of alveolar epithelial cells caused apoptosis and leakage of albumin into airways, reflecting loss of epithelial barrier function. These data establish inhalation of aerosolized virus as a critical source of exposure for fatal human infection and reveal that direct viral effects in alveoli mediate H5N1 disease. This new nonhuman primate model will advance vaccine and therapeutic approaches to prevent and treat human disease caused by highly pathogenic avian influenza viruses.
    MeSH term(s) Aerosols ; Alveolar Epithelial Cells/immunology ; Alveolar Epithelial Cells/pathology ; Alveolar Epithelial Cells/virology ; Animals ; Cells, Cultured ; Cytokines/biosynthesis ; Cytokines/immunology ; Disease Models, Animal ; Immunity, Innate/immunology ; Influenza A Virus, H5N1 Subtype/immunology ; Influenza A Virus, H5N1 Subtype/pathogenicity ; Influenza A Virus, H5N1 Subtype/physiology ; Lung/immunology ; Lung/virology ; Macaca fascicularis ; Macrophages, Alveolar/immunology ; Macrophages, Alveolar/pathology ; Macrophages, Alveolar/virology ; Orthomyxoviridae Infections/immunology ; Orthomyxoviridae Infections/physiopathology ; Orthomyxoviridae Infections/virology ; Pneumonia, Viral/immunology ; Pneumonia, Viral/virology ; Pulmonary Alveoli/virology ; Respiratory Distress Syndrome, Adult/immunology ; Respiratory Distress Syndrome, Adult/physiopathology ; Respiratory Distress Syndrome, Adult/virology ; Virus Replication
    Chemical Substances Aerosols ; Cytokines
    Language English
    Publishing date 2017-02-15
    Publishing country United States
    Document type Journal Article
    ZDB-ID 3056-9
    ISSN 1550-6606 ; 0022-1767 ; 1048-3233 ; 1047-7381
    ISSN (online) 1550-6606
    ISSN 0022-1767 ; 1048-3233 ; 1047-7381
    DOI 10.4049/jimmunol.1601770
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  7. Article: Tropism, replication competence, and innate immune responses of the coronavirus SARS-CoV-2 in human respiratory tract and conjunctiva: an analysis in ex-vivo and in-vitro cultures

    Hui, Kenrie P Y / Cheung, Man-Chun / Perera, Ranawaka A P M / Ng, Ka-Chun / Bui, Christine H T / Ho, John C W / Ng, Mandy M T / Kuok, Denise I T / Shih, Kendrick C / Tsao, Sai-Wah / Poon, Leo L M / Peiris, Malik / Nicholls, John M / Chan, Michael C W

    Lancet Respir Med

    Abstract: ... and macrophages. In-vitro studies included the highly pathogenic avian influenza H5N1 virus (H5N1) and ... and innate immune responses in comparison with other coronavirus and influenza virus to provide ... associated coronavirus (MERS-CoV), and 2009 pandemic influenza H1N1 (H1N1pdm) in ex-vivo cultures of human ...

    Abstract BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019, causing a respiratory disease (coronavirus disease 2019, COVID-19) of varying severity in Wuhan, China, and subsequently leading to a pandemic. The transmissibility and pathogenesis of SARS-CoV-2 remain poorly understood. We evaluate its tissue and cellular tropism in human respiratory tract, conjunctiva, and innate immune responses in comparison with other coronavirus and influenza virus to provide insights into COVID-19 pathogenesis. METHODS: We isolated SARS-CoV-2 from a patient with confirmed COVID-19, and compared virus tropism and replication competence with SARS-CoV, Middle East respiratory syndrome-associated coronavirus (MERS-CoV), and 2009 pandemic influenza H1N1 (H1N1pdm) in ex-vivo cultures of human bronchus (n=5) and lung (n=4). We assessed extrapulmonary infection using ex-vivo cultures of human conjunctiva (n=3) and in-vitro cultures of human colorectal adenocarcinoma cell lines. Innate immune responses and angiotensin-converting enzyme 2 expression were investigated in human alveolar epithelial cells and macrophages. In-vitro studies included the highly pathogenic avian influenza H5N1 virus (H5N1) and mock-infected cells as controls. FINDINGS: SARS-CoV-2 infected ciliated, mucus-secreting, and club cells of bronchial epithelium, type 1 pneumocytes in the lung, and the conjunctival mucosa. In the bronchus, SARS-CoV-2 replication competence was similar to MERS-CoV, and higher than SARS-CoV, but lower than H1N1pdm. In the lung, SARS-CoV-2 replication was similar to SARS-CoV and H1N1pdm, but was lower than MERS-CoV. In conjunctiva, SARS-CoV-2 replication was greater than SARS-CoV. SARS-CoV-2 was a less potent inducer of proinflammatory cytokines than H5N1, H1N1pdm, or MERS-CoV. INTERPRETATION: The conjunctival epithelium and conducting airways appear to be potential portals of infection for SARS-CoV-2. Both SARS-CoV and SARS-CoV-2 replicated similarly in the alveolar epithelium; SARS-CoV-2 replicated more extensively in the bronchus than SARS-CoV. These findings provide important insights into the transmissibility and pathogenesis of SARS-CoV-2 infection and differences with other respiratory pathogens. FUNDING: US National Institute of Allergy and Infectious Diseases, University Grants Committee of Hong Kong Special Administrative Region, China; Health and Medical Research Fund, Food and Health Bureau, Government of Hong Kong Special Administrative Region, China.
    Keywords covid19
    Publisher WHO
    Document type Article
    Note WHO #Covidence: #197584
    Database COVID19

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