scholarly journals Primary Swine Respiratory Epithelial Cell Lines for the Efficient Isolation and Propagation of Influenza A Viruses

2020 ◽  
Vol 94 (24) ◽  
Author(s):  
Victoria Meliopoulos ◽  
Sean Cherry ◽  
Nicholas Wohlgemuth ◽  
Rebekah Honce ◽  
Karen Barnard ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Epithelial Cells ◽  
Cell Lines ◽  
Influenza A ◽  
Vaccine Development ◽  
Mdck Cells ◽  
Influenza Viruses ◽  
Clinical Samples ◽  
Respiratory Epithelial Cells ◽  
Respiratory Epithelial

ABSTRACT Influenza virus isolation from clinical samples is critical for the identification and characterization of circulating and emerging viruses. Yet efficient isolation can be difficult. In these studies, we isolated primary swine nasal and tracheal respiratory epithelial cells and immortalized swine nasal epithelial cells (siNEC) and tracheal epithelial cells (siTEC) that retained the abilities to form tight junctions and cilia and to differentiate at the air-liquid interface like primary cells. Critically, both human and swine influenza viruses replicated in the immortalized cells, which generally yielded higher-titer viral isolates from human and swine nasal swabs, supported the replication of isolates that failed to grow in Madin-Darby canine kidney (MDCK) cells, and resulted in fewer dominating mutations during viral passaging than MDCK cells. IMPORTANCE Robust in vitro culture systems for influenza virus are critically needed. MDCK cells, the most widely used cell line for influenza isolation and propagation, do not adequately model the respiratory tract. Therefore, many clinical isolates, both animal and human, are unable to be isolated and characterized, limiting our understanding of currently circulating influenza viruses. We have developed immortalized swine respiratory epithelial cells that retain the ability to differentiate and can support influenza replication and isolation. These cell lines can be used as additional tools to enhance influenza research and vaccine development.

scholarly journals Amino Acid 226 in the Hemagglutinin of H4N6 Influenza Virus Determines Binding Affinity for α2,6-Linked Sialic Acid and Infectivity Levels in Primary Swine and Human Respiratory Epithelial Cells

2008 ◽  
Vol 82 (16) ◽  
pp. 8204-8209 ◽  
Author(s):  
Allen C. Bateman ◽  
Marc G. Busch ◽  
Alexander I. Karasin ◽  
Nicolai Bovin ◽  
Christopher W. Olsen
Keyword(s):  
Amino Acids ◽  
Influenza Virus ◽  
Sialic Acid ◽  
Amino Acid ◽  
Epithelial Cells ◽  
Influenza Viruses ◽  
Respiratory Epithelial Cells ◽  
Relative Binding ◽  
Human Respiratory Epithelial Cells ◽  
Respiratory Epithelial

ABSTRACT Avian lineage H4N6 influenza viruses previously isolated from pigs differ at hemagglutinin amino acids 226 and 228 from H4 subtype viruses isolated from birds. Using a parental H4N6 swine isolate and hemagglutinin mutant viruses (at residues 226 and/or 228), we determined that viruses which contain L226 had a higher affinity for sialic acid α2,6 galactose (SAα2,6Gal) and a higher infectivity level for primary swine and human respiratory epithelial cells, whereas viruses which contain Q226 had lower SAα2,6Gal affinity and lower infectivity levels for both types of cells. Using specific neuraminidases, we found that irrespective of their relative binding preferences, all of the influenza viruses examined utilized SAα2,6Gal to infect swine and human cells.

Hypothiocyanite produced by human and rat respiratory epithelial cells inactivates extracellular H1N2 influenza A virus

2015 ◽  
Vol 65 (1) ◽  
pp. 71-80 ◽  
Author(s):  
Aaron Gingerich ◽  
Lan Pang ◽  
Jarod Hanson ◽  
Daniel Dlugolenski ◽  
Rebecca Streich ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Influenza A Virus ◽  
Influenza A ◽  
Respiratory Epithelial Cells ◽  
Respiratory Epithelial

scholarly journals Influenza A Virus Protein PB1-F2 Exacerbates IFN-β Expression of Human Respiratory Epithelial Cells

2010 ◽  
Vol 185 (8) ◽  
pp. 4812-4823 ◽  
Author(s):  
Ronan Le Goffic ◽  
Edwige Bouguyon ◽  
Christophe Chevalier ◽  
Jasmina Vidic ◽  
Bruno Da Costa ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Influenza A Virus ◽  
Influenza A ◽  
Virus Protein ◽  
Respiratory Epithelial Cells ◽  
Human Respiratory Epithelial Cells ◽  
Respiratory Epithelial

scholarly journals Characterization of a Porcine Lung Epithelial Cell Line Suitable for Influenza Virus Studies

2001 ◽  
Vol 75 (19) ◽  
pp. 9517-9525 ◽  
Author(s):  
Sang Heui Seo ◽  
Olga Goloubeva ◽  
Richard Webby ◽  
Robert G. Webster
Keyword(s):  
Influenza Virus ◽  
Epithelial Cell ◽  
Cell Line ◽  
Mdck Cells ◽  
Influenza Viruses ◽  
Lung Epithelial Cell ◽  
Clinical Samples ◽  
Epithelial Cell Line ◽  
Porcine Endogenous Retrovirus ◽  
Lung Epithelial

ABSTRACT We established a porcine lung epithelial cell line designated St. Jude porcine lung cells (SJPL) and demonstrated that all tested influenza A and B viruses replicated in this cell line. The infectivity titers of most viruses in SJPL cells were comparable to or better than those in MDCK cells. The propagation of influenza viruses from clinical samples in SJPL cells did not lead to antigenic changes in the hemagglutinin molecule. The numbers of both Sia2-3Gal and Sia2-6Gal receptors on SJPL cells were greater than those on MDCK cells. Influenza virus infection of SJPL cells did not lead to apoptosis, as did infection of MDCK cells. No porcine endogenous retrovirus was detected in SJPL cells, and in contrast to MDCK cells, SJPL cells did not cause tumors in nude mice.

scholarly journals Host-directed combinatorial RNAi improves inhibition of diverse strains of influenza A virus in human respiratory epithelial cells

PLoS ONE ◽  
2018 ◽  
Vol 13 (5) ◽  
pp. e0197246 ◽  
Author(s):  
Michael A. Estrin ◽  
Islam T. M. Hussein ◽  
Wendy B. Puryear ◽  
Anne C. Kuan ◽  
Stephen C. Artim ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Influenza A Virus ◽  
Influenza A ◽  
Respiratory Epithelial Cells ◽  
Human Respiratory Epithelial Cells ◽  
Respiratory Epithelial

scholarly journals A Single-Cycle Influenza A Virus-Based SARS-CoV-2 Vaccine Elicits Potent Immune Responses in a Mouse Model

Vaccines ◽  
2021 ◽  
Vol 9 (8) ◽  
pp. 850
Author(s):  
Surapong Koonpaew ◽  
Challika Kaewborisuth ◽  
Kanjana Srisutthisamphan ◽  
Asawin Wanitchang ◽  
Theeradej Thaweerattanasinp ◽  
...  
Keyword(s):  
Influenza Virus ◽  
Immune Responses ◽  
Influenza A ◽  
Vaccine Development ◽  
Vaccine Candidate ◽  
Mdck Cells ◽  
Chimeric Gene ◽  
Cell Mediated Immunity ◽  
Single Cycle ◽  
Virus Envelope

The use of virus-vectored platforms has increasingly gained attention in vaccine development as a means for delivering antigenic genes of interest into target hosts. Here, we describe a single-cycle influenza virus-based SARS-CoV-2 vaccine designated as scPR8-RBD-M2. The vaccine utilizes the chimeric gene encoding 2A peptide-based bicistronic protein cassette of the SARS-CoV-2 receptor-binding domain (RBD) and influenza matrix 2 (M2) protein. The C-terminus of the RBD was designed to link with the cytoplasmic domain of the influenza virus hemagglutinin (HA) to anchor the RBD on the surface of producing cells and virus envelope. The chimeric RBD-M2 gene was incorporated in place of the HA open-reading frame (ORF) between the 3′ and 5′ UTR of HA gene for the virus rescue in MDCK cells stably expressing HA. The virus was also constructed with the disrupted M2 ORF in segment seven to ensure that M2 from the RBD-M2 was utilized. The chimeric gene was intact and strongly expressed in infected cells upon several passages, suggesting that the antigen was stably maintained in the vaccine candidate. Mice inoculated with scPR8-RBD-M2 via two alternative prime-boost regimens (intranasal-intranasal or intranasal-intramuscular routes) elicited robust mucosal and systemic humoral immune responses and cell-mediated immunity. Notably, we demonstrated that immunized mouse sera exhibited neutralizing activity against pseudotyped viruses bearing SARS-CoV-2 spikes from various variants, albeit with varying potency. Our study warrants further development of a replication-deficient influenza virus as a promising SARS-CoV-2 vaccine candidate.

scholarly journals Viral Coinfection Replaces Effects of Suilysin onStreptococcus suisAdherence to and Invasion of Respiratory Epithelial Cells Grown under Air-Liquid Interface Conditions

2019 ◽  
Vol 87 (8) ◽  
Author(s):  
Fandan Meng ◽  
Jie Tong ◽  
Désirée Vötsch ◽  
Ju-Yi Peng ◽  
Xuehui Cai ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Bacterial Infection ◽  
Liquid Interface ◽  
Influenza Viruses ◽  
Striking Difference ◽  
Respiratory Epithelial Cells ◽  
Content Type ◽  
Secondary Bacterial Infection ◽  
Respiratory Epithelial ◽  
Air Liquid Interface

ABSTRACTStreptococcus suisis an important zoonotic pathogen which can infect humans and pigs worldwide, posing a potential risk to global public health. Suilysin, a pore-forming cholesterol-dependent cytolysin, is considered to play an important role in the pathogenesis ofS. suisinfections. It is known that infection with influenza A viruses may favor susceptibility to secondary bacterial infection, resulting in more severe disease and increased mortality. However, the molecular mechanisms underlying these coinfections are incompletely understood. Applying highly differentiated primary porcine respiratory epithelial cells grown under air-liquid interface (ALI) conditions, we analyzed the contribution of swine influenza viruses (SIV) to the virulence ofS. suis, with a special focus on its cytolytic toxin, suilysin. We found that during secondary bacterial infection, suilysin ofS. suiscontributed to the damage of well-differentiated respiratory epithelial cells in the early stage of infection, whereas the cytotoxic effects induced by SIV became prominent at later stages of infection. Prior infection by SIV enhanced the adherence to and colonization of porcine airway epithelial cells by a wild-type (wt)S. suisstrain and a suilysin-negativeS. suismutant in a sialic acid-dependent manner. A striking difference was observed with respect to bacterial invasion. After bacterial monoinfection, only the wtS. suisstrain showed an invasive phenotype, whereas the mutant remained adherent. When the epithelial cells were preinfected with SIV, the suilysin-negative mutant also showed an invasion capacity. Therefore, we propose that coinfection with SIV may compensate for the lack of suilysin in the adherence and invasion process of suilysin-negativeS. suis.

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