Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization
Abstract The process of viral entry into host cells is crucial for the establishment of infection and the determination of viral pathogenicity. A comprehensive understanding of entry pathways is fundamental for the development of novel therapeutic strategies. Standard techniques for investigating vi...
Saved in:
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-01-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-025-87080-x |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832585777596334080 |
|---|---|
| author | Agata Kublicka Daria Lorek Agata Mikołajczyk-Martinez Grzegorz Chodaczek Aleksandra Chwirot Barbara Bażanów Anna Karolina Matczuk |
| author_facet | Agata Kublicka Daria Lorek Agata Mikołajczyk-Martinez Grzegorz Chodaczek Aleksandra Chwirot Barbara Bażanów Anna Karolina Matczuk |
| author_sort | Agata Kublicka |
| collection | DOAJ |
| description | Abstract The process of viral entry into host cells is crucial for the establishment of infection and the determination of viral pathogenicity. A comprehensive understanding of entry pathways is fundamental for the development of novel therapeutic strategies. Standard techniques for investigating viral entry include confocal microscopy and flow cytometry, both of which provide complementary qualitative and quantitative data. Imaging flow cytometry, which integrates the advantages of both methodologies, offers significant potential in virological studies. In this investigation, we employed imaging flow cytometry coupled with immunostaining to monitor the entry of equine arteritis virus EAV into Vero cells via the endosomal trafficking route. Analysis provided an insight into the early infection dynamics across thousands of cells, revealing statistically significant alterations in internalization and uncoating process. Moreover, we evaluated the effectiveness of two inhibitors targeting cellular factors involved in facilitating viral entry: ammonium chloride, which disrupts endocytosis, and camostat mesylate, which inhibits the activity of serine proteases. The results demonstrated a clear distinction between effective and ineffective inhibitors. This study highlighted the potential of imaging flow cytometry to advance the study of viral entry and the evaluation of antiviral agents. |
| format | Article |
| id | doaj-art-63b32ea95bc444baa97fb9493cc83abf |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-63b32ea95bc444baa97fb9493cc83abf2025-01-26T12:31:03ZengNature PortfolioScientific Reports2045-23222025-01-0115111310.1038/s41598-025-87080-xImaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalizationAgata Kublicka0Daria Lorek1Agata Mikołajczyk-Martinez2Grzegorz Chodaczek3Aleksandra Chwirot4Barbara Bażanów5Anna Karolina Matczuk6Department of Pathology, Division of Microbiology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesDepartment of Immunology, Pathophysiology and Veterinary Preventive Medicine, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesDepartment of Biochemistry and Molecular Biology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesLaboratory of Confocal Microscopy, Łukasiewicz Research Network – PORT Polish Center For Technology DevelopmentDepartment of Pathology, Division of Microbiology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesDepartment of Pathology, Division of Microbiology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesDepartment of Pathology, Division of Microbiology, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life SciencesAbstract The process of viral entry into host cells is crucial for the establishment of infection and the determination of viral pathogenicity. A comprehensive understanding of entry pathways is fundamental for the development of novel therapeutic strategies. Standard techniques for investigating viral entry include confocal microscopy and flow cytometry, both of which provide complementary qualitative and quantitative data. Imaging flow cytometry, which integrates the advantages of both methodologies, offers significant potential in virological studies. In this investigation, we employed imaging flow cytometry coupled with immunostaining to monitor the entry of equine arteritis virus EAV into Vero cells via the endosomal trafficking route. Analysis provided an insight into the early infection dynamics across thousands of cells, revealing statistically significant alterations in internalization and uncoating process. Moreover, we evaluated the effectiveness of two inhibitors targeting cellular factors involved in facilitating viral entry: ammonium chloride, which disrupts endocytosis, and camostat mesylate, which inhibits the activity of serine proteases. The results demonstrated a clear distinction between effective and ineffective inhibitors. This study highlighted the potential of imaging flow cytometry to advance the study of viral entry and the evaluation of antiviral agents.https://doi.org/10.1038/s41598-025-87080-xVirus entryImaging flow cytometryImFCEquine arteritis virus (EAV)Protease inhibitors |
| spellingShingle | Agata Kublicka Daria Lorek Agata Mikołajczyk-Martinez Grzegorz Chodaczek Aleksandra Chwirot Barbara Bażanów Anna Karolina Matczuk Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization Scientific Reports Virus entry Imaging flow cytometry ImFC Equine arteritis virus (EAV) Protease inhibitors |
| title | Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization |
| title_full | Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization |
| title_fullStr | Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization |
| title_full_unstemmed | Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization |
| title_short | Imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization |
| title_sort | imaging flow cytometry reveals the mechanism of equine arteritis virus entry and internalization |
| topic | Virus entry Imaging flow cytometry ImFC Equine arteritis virus (EAV) Protease inhibitors |
| url | https://doi.org/10.1038/s41598-025-87080-x |
| work_keys_str_mv | AT agatakublicka imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization AT darialorek imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization AT agatamikołajczykmartinez imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization AT grzegorzchodaczek imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization AT aleksandrachwirot imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization AT barbarabazanow imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization AT annakarolinamatczuk imagingflowcytometryrevealsthemechanismofequinearteritisvirusentryandinternalization |