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Enhanced endothelial cytopathogenicity induced by a cytomegalovirus strain propagated in endothelial cells
W J Waldman1, J M Sneddon, R E Stephens
1Department of Pathology, Ohio State University College of Medicine, Columbus.
Abstract:
Endothelial involvement has been implicated in cytomegalovirus (CMV) infection, a source of major complications in immunosuppressed individuals (e.g., those with acquired immune deficiency syndrome [AIDS] and organ transplants). Traditionally, CMV has been grown in fibroblasts; however, propagation in these cells may alter characteristics of the virus. In developing an in vitro model system of CMV/endothelial cell interaction, we have addressed this issue by propagating a clinical isolate, CMV VHL 1, in human umbilical vein endothelial (HUVE) cells by serial cocultivation of heavily infected cultures with fresh HUVE monolayers and have compared its infectious properties with those of the fibroblast-raised strain, CMV AD169. In situ hybridization using a biotinylated DNA probe, as well as immunofluorescent staining for CMV-specific antigen, has confirmed infection of HUVE cells inoculated with either strain of the virus. Infection of HUVE by VHL was accompanied by dramatic cytopathology not observed in AD169-infected cells. Plaque assay of culture supernatants revealed greater virus production in VHL-infected HUVE as compared with equivalently inoculated fibroblasts. In contrast, AD169 production in inoculated fibroblasts exceeded that in HUVE. These studies demonstrate the suitability of cultured endothelial cells as a substrate for CMV propagation and suggest that a strain of virus thus propagated may offer an accurate model of CMV/endothelial cell interaction in human disease.
Insights
Cytomegalovirus (CMV) can infect endothelial cells, causing complications in immunocompromised patients. Propagating CMV in human umbilical vein endothelial (HUVE) cells provides a more accurate model for studying CMV-endothelial cell interactions in disease.
Area of Science:
- Virology
- Cell Biology
- Immunology
Background:
- Endothelial cell involvement is critical in cytomegalovirus (CMV) infections, particularly in immunocompromised individuals like organ transplant recipients and those with acquired immune deficiency syndrome (AIDS).
- Traditional CMV propagation in fibroblasts may alter viral characteristics, potentially limiting its utility as a disease model.
- Developing an in vitro model using endothelial cells is crucial for accurately studying CMV-endothelial cell interactions.
Purpose of the Study:
- To establish an in vitro model of CMV/endothelial cell interaction by propagating a clinical CMV isolate in human umbilical vein endothelial (HUVE) cells.
- To compare the infectious properties of a fibroblast-grown CMV strain (AD169) with a strain propagated in HUVE cells (VHL 1).
Main Methods:
- Serial cocultivation of a clinical CMV isolate (VHL 1) with fresh HUVE monolayers to propagate the virus.
- Comparison of CMV VHL 1 propagated in HUVE cells with CMV AD169 propagated in fibroblasts.
- Confirmation of HUVE cell infection using in situ hybridization and immunofluorescent staining for CMV-specific antigen.
- Assessment of virus production via plaque assays in infected HUVE cells and fibroblasts.
Main Results:
- Both CMV VHL 1 and CMV AD169 successfully infected HUVE cells, confirmed by molecular and immunological assays.
- CMV VHL 1 propagated in HUVE cells induced significant cytopathology, unlike CMV AD169.
- Virus production was higher for VHL 1 in HUVE cells compared to fibroblasts, while AD169 production was greater in fibroblasts than in HUVE cells.
Conclusions:
- Cultured endothelial cells are a suitable substrate for propagating CMV, offering a more relevant model.
- A CMV strain propagated in endothelial cells may more accurately reflect CMV-endothelial cell interactions observed in human diseases.
- This model system is valuable for understanding CMV pathogenesis in endothelial cells, particularly in immunocompromised states.
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