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Kinetics of BoHV-1 dissemination in an in vitro culture of bovine upper respiratory tract mucosa explants

Lennert Steukers (UGent) , Annelies Vandekerckhove (UGent) , Wim Van Den Broeck (UGent) , Sarah Glorieux (UGent) and Hans Nauwynck (UGent)
(2012) ILAR JOURNAL. 53(1). p.E43-E54
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Abstract
Bovine herpesvirus type 1 (BoHV-1) is a well known disease-causing agent in cattle. Detailed information on viral behavior with emphasis on host invasion at primary replication sites such as the mucosa of the upper respiratory tract are still scarce. Therefore, an in vitro system of bovine upper respiratory tract (bURT) mucosa explants was set up to study BoHV-1 molecular/cellular host-pathogen interactions. A thorough morphometrical analysis (epithelial integrity, basement membrane (BM) continuity and lamina propria integrity) was performed by means of light microscopy and transmission electron microscopy. A Terminal deoxynucleotidyl transferase mediated dUTP Nick End Labelling (TUNEL) staining as viability test was applied. bURT mucosa explants were maintained in culture for up to 96h without any significant changes regarding morphometry and viability. Next, bURT mucosa explants were infected with BoHV-1 (Cooper) and collected at 0h, 24h, 48h and 72h post inoculation (pi). An assessment of dissemination characteristics, using a quantitative analysis system to measure plaque latitude and plaque invasion depth, in relation to elapsed time pi was performed. A plaquewise spread of BoHV-1 across the BM as soon as 24h pi was witnessed, similar to pseudorabies virus (PrV). Moreover, BoHV-1 was found to exhibit an increased capacity to invade in proximal tracheal tissues compared to tissues of the deeper part of the nasal septum and ventral conchae. Revealing already a more distinct invasion of BoHV-1 in proximal trachea, we can conclude that the bURT mucosa explant model is suitable to study an important aspect of BoHV-1 pathogenesis.
Keywords
in vitro model, pathogenesis, bovine upper respiratory tract (bURT), bovine herpesvirus 1 (BoHV-1), basement membrane, morphometry, terminal deoxynucleotidyl transferase mediated dUTP nick end labeling, upper respiratory mucosa, viability, TRACHEAL ORGAN-CULTURES, HERPESVIRUS TYPE-1, NASAL-MUCOSA, MOLECULAR CHARACTERISTICS, RHINOTRACHEITIS VIRUS, REPLICATION KINETICS, DRUG TRANSPORT, CELL-CULTURES, BASAL CELLS, INFECTION

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Chicago
Steukers, Lennert, Annelies Vandekerckhove, Wim Van Den Broeck, Sarah Glorieux, and Hans Nauwynck. 2012. “Kinetics of BoHV-1 Dissemination in an in Vitro Culture of Bovine Upper Respiratory Tract Mucosa Explants.” Ilar Journal 53 (1): E43–E54.
APA
Steukers, L., Vandekerckhove, A., Van Den Broeck, W., Glorieux, S., & Nauwynck, H. (2012). Kinetics of BoHV-1 dissemination in an in vitro culture of bovine upper respiratory tract mucosa explants. ILAR JOURNAL, 53(1), E43–E54.
Vancouver
1.
Steukers L, Vandekerckhove A, Van Den Broeck W, Glorieux S, Nauwynck H. Kinetics of BoHV-1 dissemination in an in vitro culture of bovine upper respiratory tract mucosa explants. ILAR JOURNAL. 2012;53(1):E43–E54.
MLA
Steukers, Lennert, Annelies Vandekerckhove, Wim Van Den Broeck, et al. “Kinetics of BoHV-1 Dissemination in an in Vitro Culture of Bovine Upper Respiratory Tract Mucosa Explants.” ILAR JOURNAL 53.1 (2012): E43–E54. Print.
@article{1153271,
  abstract     = {Bovine herpesvirus type 1 (BoHV-1) is a well known disease-causing agent in cattle. Detailed information on viral behavior with emphasis on host invasion at primary replication sites such as the mucosa of the upper respiratory tract are still scarce. Therefore, an in vitro system of bovine upper respiratory tract (bURT) mucosa explants was set up to study BoHV-1 molecular/cellular host-pathogen interactions. A thorough morphometrical analysis (epithelial integrity, basement membrane (BM) continuity and lamina propria integrity) was performed by means of light microscopy and transmission electron microscopy. A Terminal deoxynucleotidyl transferase mediated dUTP Nick End Labelling (TUNEL) staining as viability test was applied. bURT mucosa explants were maintained in culture for up to 96h without any significant changes regarding morphometry and viability. Next, bURT mucosa explants were infected with BoHV-1 (Cooper) and collected at 0h, 24h, 48h and 72h post inoculation (pi). An assessment of dissemination characteristics, using a quantitative analysis system to measure plaque latitude and plaque invasion depth, in relation to elapsed time pi was performed. A plaquewise spread of BoHV-1 across the BM as soon as 24h pi was witnessed, similar to pseudorabies virus (PrV). Moreover, BoHV-1 was found to exhibit an increased capacity to invade in proximal tracheal tissues compared to tissues of the deeper part of the nasal septum and ventral conchae. Revealing already a more distinct invasion of BoHV-1 in proximal trachea, we can conclude that the bURT mucosa explant model is suitable to study an important aspect of BoHV-1 pathogenesis.},
  author       = {Steukers, Lennert and Vandekerckhove, Annelies and Van Den Broeck, Wim and Glorieux, Sarah and Nauwynck, Hans},
  issn         = {1084-2020},
  journal      = {ILAR JOURNAL},
  keyword      = {in vitro model,pathogenesis,bovine upper respiratory tract (bURT),bovine herpesvirus 1 (BoHV-1),basement membrane,morphometry,terminal deoxynucleotidyl transferase mediated dUTP nick end labeling,upper respiratory mucosa,viability,TRACHEAL ORGAN-CULTURES,HERPESVIRUS TYPE-1,NASAL-MUCOSA,MOLECULAR CHARACTERISTICS,RHINOTRACHEITIS VIRUS,REPLICATION KINETICS,DRUG TRANSPORT,CELL-CULTURES,BASAL CELLS,INFECTION},
  language     = {eng},
  number       = {1},
  pages        = {E43--E54},
  title        = {Kinetics of BoHV-1 dissemination in an in vitro culture of bovine upper respiratory tract mucosa explants},
  url          = {http://dx.doi.org/10.1093/ilar.53.1.43},
  volume       = {53},
  year         = {2012},
}

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