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Updated: May 10, 2026
An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection
Published on: December 10, 2013
Paramyxovirus infections in ex vivo lung slice cultures of different host species
D Tien Nguyen1, Rory D de Vries, Martin Ludlow
1Department of Viroscience, Erasmus MC, University Medical Center, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
Abstract:
Paramyxoviruses, including measles virus (MV), human metapneumovirus (HMPV), human respiratory syncytial virus (HRSV) and canine distemper virus (CDV), are transmitted via the respiratory route. Despite their close phylogenetic relationship, the pathogenesis of these viruses is very different. To study viral tropism and replication ex vivo, a protocol for the inflation of lungs with low-melting-point agarose mixed with culture medium was established. Lung slices were prepared and remained viable in culture at 37°C for at least 7 days. Lung slices obtained from different animal species were infected with recombinant paramyxoviruses expressing enhanced green fluorescent protein (EGFP). Progression of infection was monitored in real time by detection of EGFP fluorescence. Flow cytometric analysis of cells emigrating into the culture medium proved to be a valuable tool for qualitative and quantitative assessment of infection over time. MV replicated optimally in lung slices of macaques, HMPV and HRSV in lung slices of cotton rats, and CDV in lung slices obtained from dogs and ferrets. In conclusion, these ex vivo lung slice cultures are discriminatory models for the study of respiratory virus infections and can be used to inform the design of future in vivo experiments, thereby reducing numbers of animals required.
Insights
This study introduces an ex vivo lung slice culture method to study respiratory virus tropism. This approach effectively differentiates viral tropism and replication across various paramyxoviruses, reducing the need for animal models.
Area of Science:
- Virology
- Respiratory Medicine
- Animal Models
Background:
- Paramyxoviruses (measles virus, human metapneumovirus, human respiratory syncytial virus, canine distemper virus) are respiratory pathogens with diverse pathogenesis.
- Understanding viral tropism and replication is crucial for developing effective treatments and vaccines.
Purpose of the Study:
- To establish and validate an ex vivo lung slice culture system for studying respiratory virus tropism and replication.
- To assess the discriminatory capacity of this model for different paramyxoviruses across various animal species.
Main Methods:
- A protocol for inflating lungs with agarose-medium mixture to create viable lung slices for ex vivo culture.
- Infection of lung slices from different species with recombinant enhanced green fluorescent protein (EGFP)-expressing paramyxoviruses.
- Real-time monitoring of infection via EGFP fluorescence and quantitative assessment using flow cytometry.
Main Results:
- Lung slices remained viable ex vivo for at least 7 days.
- Measles virus showed optimal replication in macaque lung slices.
- Human metapneumovirus and human respiratory syncytial virus replicated best in cotton rat lung slices.
- Canine distemper virus replicated optimally in dog and ferret lung slices.
Conclusions:
- Ex vivo lung slice cultures serve as discriminatory models for studying respiratory virus infections.
- This model can inform in vivo experimental design, potentially reducing animal usage.
- The system accurately predicts species-specific tropism for various paramyxoviruses.
