Equine rhinitis A virus infection and cytokine expression in primary tracheobronchial epithelial cell culture
Bibliographic record
Abstract
Primary bronchial cells and ex-vivo explants are commonly used as a model for respiratory viral infections and lung diseases in humans. Similarly, these systems can be used to mimic and study respiratory conditions in the horse. Therefore, equine tracheobronchial epithelial cells (TBEC) could be used to investigate viral respiratory infections and their immune/inflammatory responses in this species. The aims of the present study were to investigate the cytopathic characteristics of equine rhinitis A virus (ERAV) in primary TBECs and the gene expression of IFN-gamma, IFN-Beta, IL-4, and IL-8. TBECs were obtained from the lower trachea, carina and primary bronchi of five euthanized horses. After dissection, the tracheal mucosa was removed, washed and kept overnight on pronase. Twelve to eighteen hours later, the airway mucosa was scraped and cells were mechanically separated. TBECs were then washed, plated on collagen coated flasks/plates using modified DMEM F12 cell culture medium and allowed to propagate to confluency. In order to investigate the cytopathic characteristics, confluent TBEC monolayers were exposed to ERAV and observed daily for 5 consecutive days. Additionally, to investigate cytokine expression by qPCR, TBEC monolayers were exposed to either ERAV or equine influenza virus (EIV) and supernatant samples were collected at 0, 2, 4, 6, 8, 10, 12, 16, 20 and 24 hours post-infection. EIV was used as a comparison for cytokine expression. Interestingly, in this study ERAV infected TBECs developed syncytial cell formations and/or cell clumping commonly observed during other in-vitro viral infections, such as syncytial or herpesviruses. Contrary to our expectations, significant changes in the expression of interferons and IL-4 were not detectable by qPCR after exposure of TBECs to ERAV or EIV within 24 hours. However, up-regulation of IL-8 after exposure to these two viruses was identified from 2 up to 24 hours post-infection in a similar magnitude. IL-8 is consistently detected in respiratory secretions during viral infections in humans and has been shown to induce chemotaxis and phagocytosis of inflammatory cells in the airways. Therefore, changes in the expression of IL-8 indicate that this chemokine might play an important role during early infections, perhaps as a chemotactic factor and/or phagocytic inductor.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.001 | 0.001 |
| Bibliometrics | 0.001 | 0.001 |
| Science and technology studies | 0.001 | 0.001 |
| Scholarly communication | 0.001 | 0.001 |
| Open science | 0.000 | 0.001 |
| Research integrity | 0.001 | 0.002 |
| Insufficient payload (model declined to judge) | 0.003 | 0.002 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".