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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
Growth of respiratory syncytial virus in mink lung epithelial cells
L R Yeolekar1, R G Damle, A Basu
1National Institute of Virology (ICMR), Pune, India.
Abstract:
Mink lung epithelial cells (Mv-1-Lu) were tested for their ability to support the growth and serial passage of respiratory syncytial virus (RSV) in vitro. Indian isolates of RSV induced distinctive cytopathic effect with typical rounding of cells followed by detachment with more than 50 per cent cells showing bright fluorescence using anti-RSV monoclonal antibodies in immunofluorescence test. Serial passage of RSV was possible in Mv-1-Lu cells without loss of sensitivity of the cells for virus growth. Titration of cell associated virus and virus released in the supernatant indicated that 60 per cent of the virus was released in the supernatant, and 40 per cent remained cell associated. Transmission electron microscopic studies of negatively stained RSV particles and ultra-thin sections of RSV infected Mv-1-Lu cells showed roughly spherical particles with club shaped projections, budding from the cytoplasmic membrane. These results indicate that Mv-1-Lu cell line is suitable for the growth and propagation of RSV.
Insights
Mink lung epithelial cells (Mv-1-Lu) effectively support respiratory syncytial virus (RSV) growth and propagation in vitro. This study confirms Mv-1-Lu cells are a suitable model for RSV research.
Area of Science:
- Virology
- Cell Biology
Background:
- Respiratory Syncytial Virus (RSV) is a significant human respiratory pathogen.
- In vitro models are crucial for studying RSV replication and developing antiviral strategies.
Purpose of the Study:
- To evaluate the suitability of the Mv-1-Lu cell line for the in vitro growth and serial passage of RSV.
- To characterize the cytopathic effects and viral release kinetics of RSV in Mv-1-Lu cells.
Main Methods:
- Infection of Mv-1-Lu cells with Indian RSV isolates.
- Assessment of cytopathic effects (CPE) via microscopy and immunofluorescence.
- Serial passage of RSV in Mv-1-Lu cells.
- Quantification of cell-associated and released virus.
- Transmission electron microscopy (TEM) for ultrastructural analysis.
Main Results:
- RSV induced characteristic CPE in Mv-1-Lu cells, including cell rounding and detachment.
- Immunofluorescence confirmed RSV infection in over 50% of cells.
- Successful serial passage of RSV without loss of viral sensitivity was achieved.
- Approximately 60% of the virus was released into the supernatant, with 40% remaining cell-associated.
- TEM revealed RSV particles with characteristic projections budding from the cell membrane.
Conclusions:
- The Mv-1-Lu cell line is a robust and suitable model for the in vitro cultivation and propagation of RSV.
- This cell line facilitates detailed studies of RSV replication, pathogenesis, and antiviral efficacy.
