Related Experiment Videos
[Analysis of the cellular tropism of human polyoma JC virus (JCV)]
1Second Department of Pathology, Hokkaido University School of Medicine, Sapporo, Japan.
Abstract:
Human polyomavirus JC virus (JCV) is the causative agent of the demyelinating disorder progressive multifocal leukoencephalopathy (PML). In vivo the cellular tropism of JCV has been shown to be very narrow, and replication appears to be essentially restricted to oligodendrocytes. To investigate the detail cellular tropism of JCV, we employed transfection, microinjection and CAT assays using JCV permissive cells, and several non-permissive cell lines. IMR-32 (human neuroblastoma cells) was permissive for IMR-32 adapted JC virus. A431 (human epidermoid carcinoma) and COS-7 (a SV40 transformed African green monkey kidney cell line) were used as non-permissive cells. Employing infection it could be confirmed that the virus proliferated in IMR-32, but not in A431 and COS-7 cells as measured by immunofluorescence methods. However, after microinjection of IMR-32 adapted JCV it could be shown that virus could replicate not only in IMR-32 but also in COS-7 cells. Virus could not be replicated in A431 cells. Using CAT assays the regulatory region of IMR-32 adapted JCV was shown to be active in IMR-32 and COS-7 cells, but inactive in A431 cells. The result suggests that nuclear transcription factors are also determinant of JCV cell tropism in vivo in addition to specific cellular receptors.
Insights
JC virus (JCV) causes progressive multifocal leukoencephalopathy (PML). This study reveals that nuclear transcription factors, not just cell receptors, influence JCV
Area of Science:
- Virology
- Neuroscience
- Cell Biology
Background:
- Human polyomavirus JC virus (JCV) causes progressive multifocal leukoencephalopathy (PML), a severe demyelinating disease.
- JCV's in vivo cellular tropism is narrow, primarily affecting oligodendrocytes.
Purpose of the Study:
- To investigate the detailed cellular tropism of JCV.
- To identify factors beyond cellular receptors that determine JCV's cell specificity.
Main Methods:
- Utilized transfection, microinjection, and CAT assays with permissive (IMR-32) and non-permissive (A431, COS-7) cell lines.
- Assessed JCV replication via immunofluorescence and regulatory region activity using CAT assays.
Main Results:
- JCV replicated in IMR-32 cells upon infection and in both IMR-32 and COS-7 cells after microinjection.
- JCV did not replicate in A431 cells under any tested condition.
- The regulatory region of JCV was active in IMR-32 and COS-7 cells but inactive in A431 cells.
Conclusions:
- Cellular receptors are not the sole determinants of JCV cell tropism.
- Nuclear transcription factors play a crucial role in dictating JCV's cellular specificity in vivo.