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Rhinovirus C Infection in Swine Respiratory Explant Cultures
Description
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A4720 - Rhinovirus C Infection in Swine Respiratory Explant Cultures
Author Block: W. Chan, Q. Tian, K. Tao, H. Tsui, T. Li, T. Leung; Paediatrics, The Chinese University of Hong Kong, Hong Kong, Hong Kong.
RATIONALE
Rhinovirus (RV) is the dominant infectious cause of hospitalization in the paediatric population. It is also the most frequently detected pathogen for the community acquired pneumonia in the adult. RV initiates acute respiratory infection and certain of them contribute to the exacerbations of underlying respiratory diseases. The epidemiology evidence indicates that, RV species C comprises at least half of the overall RV detected in kids and has a stronger association to childhood asthma exacerbations and is more associated to viremia cases than the summation of the RV-A and RV-B. Yet, there is neither an effective antiviral nor a vaccine against RVs. In the past 50 years, we learnt much about RV species A and B. In contrast, RV-C is understudied. The identification of an RV-C susceptible animal model would be the initial step for us to understand the acute infection and the long-term outcomes of RV-C.
METHODS
The intact respiratory organ of 6-month-old pigs were obtained. The tissue explants of tracheal, bronchial and alveolar epithelia were prepared as described. RV-A2 (minor), RV-B14 (major), and RV-C15 were constructed by reverse genetic method with an insertion of eGFP into the full-length RV-A2, RV-B14, and RV-C15 infectious cDNA in the pMJ3 cloning vector (gifts from Dr Lee WM). The respiratory explant cultures were washed to remove the possible cytokine secreted due to the preparation process. The four eGFP-RVs were inoculated at 105 TCID50/ml with a negative control by submerging the tissue snippets into 1 ml of virus stock. At 1, 24, 48, 72 and 120 hours post infection (hpi), viral gene quantification and viral load titration in were performed to study the viral kinetics.
RESULTS
The RV-A2 and RV-C15 replicated in the tracheal explant in the first 24 hours with 950,000 and 580,000 viral gene copies detected, respectively. RV-B14 had a slower kinetics with 600,000 vial gene copies detected at 72 hpi. Interestingly, the lung served as the susceptible site for RV-C15 replication. We detected the highest viral count at 72 hpi upon the challenge of RV-C15 in lung when compared to the RV-A2 and RV-B14.
CONCLUSION
We hypothesize that pig respiratory organ explant culture is susceptible to RV-C infection and replication and it would serve as a reliable predicator for their susceptibility in vivo. The result of this project could be used as an initial platform for rhinovirus study and drug screening.
Author Block: W. Chan, Q. Tian, K. Tao, H. Tsui, T. Li, T. Leung; Paediatrics, The Chinese University of Hong Kong, Hong Kong, Hong Kong.
RATIONALE
Rhinovirus (RV) is the dominant infectious cause of hospitalization in the paediatric population. It is also the most frequently detected pathogen for the community acquired pneumonia in the adult. RV initiates acute respiratory infection and certain of them contribute to the exacerbations of underlying respiratory diseases. The epidemiology evidence indicates that, RV species C comprises at least half of the overall RV detected in kids and has a stronger association to childhood asthma exacerbations and is more associated to viremia cases than the summation of the RV-A and RV-B. Yet, there is neither an effective antiviral nor a vaccine against RVs. In the past 50 years, we learnt much about RV species A and B. In contrast, RV-C is understudied. The identification of an RV-C susceptible animal model would be the initial step for us to understand the acute infection and the long-term outcomes of RV-C.
METHODS
The intact respiratory organ of 6-month-old pigs were obtained. The tissue explants of tracheal, bronchial and alveolar epithelia were prepared as described. RV-A2 (minor), RV-B14 (major), and RV-C15 were constructed by reverse genetic method with an insertion of eGFP into the full-length RV-A2, RV-B14, and RV-C15 infectious cDNA in the pMJ3 cloning vector (gifts from Dr Lee WM). The respiratory explant cultures were washed to remove the possible cytokine secreted due to the preparation process. The four eGFP-RVs were inoculated at 105 TCID50/ml with a negative control by submerging the tissue snippets into 1 ml of virus stock. At 1, 24, 48, 72 and 120 hours post infection (hpi), viral gene quantification and viral load titration in were performed to study the viral kinetics.
RESULTS
The RV-A2 and RV-C15 replicated in the tracheal explant in the first 24 hours with 950,000 and 580,000 viral gene copies detected, respectively. RV-B14 had a slower kinetics with 600,000 vial gene copies detected at 72 hpi. Interestingly, the lung served as the susceptible site for RV-C15 replication. We detected the highest viral count at 72 hpi upon the challenge of RV-C15 in lung when compared to the RV-A2 and RV-B14.
CONCLUSION
We hypothesize that pig respiratory organ explant culture is susceptible to RV-C infection and replication and it would serve as a reliable predicator for their susceptibility in vivo. The result of this project could be used as an initial platform for rhinovirus study and drug screening.
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