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Using a Phosphoproteomic Screen to Profile Early Changes During HCMV Infection of Human Monocytes
Liudmila S Chesnokova1, Andrew D Yurochko2
1Department of Microbiology & Immunology, Center for Molecular and Tumor Virology, Feist-Weiller Cancer Center, Louisiana State University Health Sciences Center-Shreveport, Shreveport, LA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|February 8, 2021
Summary
Human cytomegalovirus (HCMV) infection involves rapid cell signaling. This study developed an ELISA-based assay to detect early phosphorylation events in monocytes during HCMV infection, aiding understanding of viral entry.
Area of Science:
- Virology and Immunology
- Cellular Signaling and Phosphoproteomics
Background:
- Human cytomegalovirus (HCMV) binds to monocytes via epidermal growth factor receptor (EGFR) and integrins, initiating rapid downstream signaling pathways.
- Understanding HCMV-induced signaling is crucial for elucidating its infection mechanisms and effects on target cell biology.
- Early phosphorylation events following HCMV receptor engagement are key indicators of cellular response.
Purpose of the Study:
- To assess rapid phosphorylation events in monocytes upon HCMV infection using an ELISA-based antibody platform.
- To investigate the phosphoproteomic landscape associated with HCMV-receptor interactions.
- To establish a versatile assay for studying early signaling events in various infection models.
Main Methods:
- Utilized an ELISA-based antibody platform for high-throughput screening of phosphorylation.
- Performed a phosphoproteomic screen using an ELISA-based antibody array.
- Focused on detecting early phosphorylation events occurring shortly after HCMV infection of monocytes.
Main Results:
- Successfully detected rapid phosphorylation events in monocytes following HCMV infection.
- Identified potential phosphorylated proteins involved in modulating HCMV infection.
- Demonstrated the utility of the ELISA-based antibody platform for phosphoproteomic analysis.
Conclusions:
- The developed ELISA-based assay effectively captures early phosphorylation events critical for HCMV infection.
- This phosphoproteomic approach provides insights into HCMV-host cell interactions.
- The assay is adaptable for diverse research applications in virology and cell signaling.
