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MCMV Centrifugal Enhancement: A New Spin on an Old Topic
Trevor J Hancock1, Morgan Lynn Hetzel1, Andrea Ramirez1
1Department of Microbiology, University of Tennessee, Knoxville, TN 37996, USA.
Abstract:
Human cytomegalovirus (HCMV) is a ubiquitous pathogen infecting a majority of people worldwide, with diseases ranging from mild to life-threatening. Its clinical relevance in immunocompromised people and congenital infections have made treatment and vaccine development a top priority. Because of cytomegaloviruses' species specificity, murine cytomegalovirus (MCMV) models have historically informed and advanced translational CMV therapies. Using the phenomenon of centrifugal enhancement, we explored differences between MCMVs derived in vitro and in vivo. We found centrifugal enhancement on tissue culture-derived virus (TCV) was ~3× greater compared with salivary gland derived virus (SGV). Using novel "flow virometry", we found that TCV contained a distinct submicron particle composition compared to SGV. Using an inhibitor of exosome production, we show these submicron particles are not extracellular vesicles that contribute to centrifugal enhancement. We examined how these differences in submicron particles potentially contribute to differing centrifugal enhancement phenotypes, as well as broader in vivo vs. in vitro MCMV differences.
Insights
Murine cytomegalovirus (MCMV) derived in vitro shows significantly higher centrifugal enhancement than in vivo-derived virus, due to distinct submicron particle composition. These findings offer insights into MCMV pathogenesis and therapeutic development.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Human cytomegalovirus (HCMV) is a widespread pathogen with significant health implications, especially for immunocompromised individuals and in congenital infections.
- Murine cytomegalovirus (MCMV) serves as a crucial model for studying HCMV due to species specificity, aiding translational therapy research.
- Understanding in vitro versus in vivo MCMV differences is vital for refining therapeutic strategies.
Purpose of the Study:
- To investigate the differences in centrifugal enhancement between MCMV derived in vitro (tissue culture-derived virus, TCV) and in vivo (salivary gland-derived virus, SGV).
- To characterize the particle composition of TCV and SGV using novel methods and explore the nature of observed differences.
Main Methods:
- Comparative analysis of centrifugal enhancement in TCV and SGV.
- Utilizing "flow virometry" to analyze the submicron particle composition of MCMV isolates.
- Employing an exosome production inhibitor to determine the role of extracellular vesicles in centrifugal enhancement.
Main Results:
- Tissue culture-derived virus (TCV) exhibited approximately threefold greater centrifugal enhancement compared to salivary gland-derived virus (SGV).
- Flow virometry revealed distinct submicron particle compositions between TCV and SGV.
- Inhibiting exosome production demonstrated that these submicron particles are not extracellular vesicles contributing to centrifugal enhancement.
Conclusions:
- Differences in submicron particle composition between in vitro and in vivo MCMV contribute to variations in centrifugal enhancement.
- These findings provide a deeper understanding of MCMV biology and highlight potential targets for therapeutic interventions.
- Further research is warranted to elucidate the precise role of these submicron particles in MCMV pathogenesis.
