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Rhinovirus infection in human and mouse precision cut lung slices

: Rochlitzer, Sabine; Müller, Meike; Sewald, Katherina; Braun, Armin

American Journal of Respiratory and Critical Care Medicine 189 (2014), Art. A1712
ISSN: 1073-449X
ISSN: 0003-0805
ISSN: 1535-4970
American Thoracic Society (ATS International Conference) <2014, San Diego/Calif.>
Fraunhofer ITEM ()

Human rhinovirus (HRV) is the main cause of respiratory tract infection and has been linked to exacerbations of chronic lung diseases e.g. allergic asthma. However, reliable models of HRV induced exacerbations of severe chronic asthma are not available. The aim of this study was to evaluate the potential use of precision cut lung slices (PCLS) for analysis of HRV infection in mouse and human.
Viable PCLS containing small airways were prepared from mouse and human lung tissue and infected with HRV1B (107 TCID /ml) or 50 UV-inactivated HRV1B as a control. Virus titre in the supernatant was measured after 24h and 48h by dilution plating on HeLa cells. HRV1B localization in the tissue was assessed by immunofluorescence staining using anti-dsRNA antibody in combination with cytokeratin staining of epithelial cells.
Only small amounts of virus were detectable in either murine or human PCLS supernatant at 24h and this further decreased at 48h thus not indicating a replication-induced release of virions. However, immunofluorescence analysis revealed a dsRNA positive staining demonstrating the presence of replicating virus. This was not observed in the UV-inactivated virus control. The specific dsRNA staining was localized to the epithelial cell layer of small airways and was present in both human and mouse PCLS.
As both mouse and human PCLS showed evidence for HRV infection, this suggests that PCLS can be used to investigate and compare viral infection in different species. Future studies will focus on the viral-induced immune response as well as the establishment of "asthmatic phenotype" PCLS to investigate potential exacerbation mechanisms.