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Published on: February 6, 2017
Productive infection of human endometrial stromal cells by human cytomegalovirus
T F Kowalik1, A D Yurochko, C A Rinehart
1Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill 27599-7295.
Abstract:
Cultured endometrial stromal cells were susceptible to productive human cytomegalovirus (HCMV) infection. Infection of endometrial stromal cells resulted in pronounced cytopathic effects including cell rounding and aggregation, fusions, and some lysis, although not in the synchronous fashion observed in infected fibroblasts. The aggregation events were reminiscent of normal endometrial stromal cell responses to cyclical estrogen/progesterone levels. Immunofluorescence analysis demonstrated expression of viral gene products suggesting a productive virus infection. One-step growth analysis showed that infectious virus was produced but the titers were two logs lower than those obtained in fibroblasts even though HCMV DNA accumulated to similar levels in both cell types. In contrast, viral DNA replication was greatly reduced in endometrial stromal cells immortalized with a temperature-sensitive SV40 large T gene at both permissive and nonpermissive temperatures. A more detailed analysis of viral gene expression by Northern blotting revealed earlier appearances and greater initial levels of viral transcripts in endometrial stromal cells. No HCMV gene expression was observed at 120 hpi in these cells even though half of the cells were still intact and cellular gene expression was functional. Since this was the time of peak virus production, it seems plausible that reduced viral gene expression at late times p.i. was a major contributor to the reduced titers observed in endometrial stromal cells. These in vitro results coupled with in vivo observations by others of endometritis associated with HCMV suggest that further investigation into the effects of HCMV on the endometrium is warranted.
Insights
Human cytomegalovirus (HCMV) productively infects endometrial stromal cells, causing cytopathic effects. Reduced viral gene expression at late infection stages contributes to lower infectious virus production compared to fibroblasts.
Area of Science:
- Gynecology
- Virology
- Cell Biology
Background:
- Human cytomegalovirus (HCMV) is a common pathogen with potential reproductive health implications.
- The endometrium, a key tissue in reproductive health, has not been extensively studied for HCMV susceptibility.
- Understanding HCMV interactions with endometrial cells is crucial for reproductive health research.
Purpose of the Study:
- To investigate the susceptibility of human endometrial stromal cells to HCMV infection.
- To characterize the cellular response and viral replication dynamics within these cells.
- To explore the potential impact of HCMV on endometrial function.
Main Methods:
- Primary human endometrial stromal cells were cultured and infected with HCMV.
- Cytopathic effects were assessed via microscopy.
- Viral gene product expression was analyzed using immunofluorescence.
- Viral replication kinetics were determined by one-step growth analysis.
- Viral DNA replication and gene expression were further analyzed in immortalized cells and by Northern blotting.
Main Results:
- Endometrial stromal cells supported productive HCMV infection with observable cytopathic effects.
- Infectious virus titers were lower in stromal cells compared to fibroblasts, despite similar viral DNA accumulation.
- Viral DNA replication was significantly reduced in immortalized stromal cells.
- Early viral gene expression was enhanced, but late-stage expression was diminished in stromal cells.
- HCMV infection did not lead to complete cellular lysis, and cellular gene expression remained functional.
Conclusions:
- Endometrial stromal cells are susceptible to productive HCMV infection, exhibiting unique replication dynamics.
- Reduced late-stage viral gene expression appears to be a key factor limiting infectious virus production.
- These findings warrant further investigation into HCMV's role in endometrial pathology, such as endometritis.
