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Comparative Assessment of Feline and Canine Coronavirus-Induced Modifications in Extracellular Vesicle Biogenesis and Composition in Host Cells /

Record Type:	Electronic resources : Monograph/item
Title/Author:	Comparative Assessment of Feline and Canine Coronavirus-Induced Modifications in Extracellular Vesicle Biogenesis and Composition in Host Cells // Sandani Vimukthika Thurumal Wijerathne.
Author:	Wijerathne, Sandani Vimukthika Thurumal,
Description:	1 electronic resource (126 pages)
Notes:	Source: Dissertations Abstracts International, Volume: 86-08, Section: B.
Contained By:	Dissertations Abstracts International86-08B.
Subject:	Microbiology. -
Online resource:	https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=31841398
ISBN:	9798304996518

Comparative Assessment of Feline and Canine Coronavirus-Induced Modifications in Extracellular Vesicle Biogenesis and Composition in Host Cells /
Wijerathne, Sandani Vimukthika Thurumal,

Comparative Assessment of Feline and Canine Coronavirus-Induced Modifications in Extracellular Vesicle Biogenesis and Composition in Host Cells /Sandani Vimukthika Thurumal Wijerathne. - 1 electronic resource (126 pages)

Source: Dissertations Abstracts International, Volume: 86-08, Section: B.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a highly contagious infectious disease that can develop from mild to severe respiratory illnesses in humans and has rapidly spread worldwide, leading to a global pandemic. The extremely pathogenic coronavirus (CoV) outbreak of SARS-CoV-2 and Middle East respiratory syndrome coronavirus (MERS-CoV) originated from animals. Numerous coronaviruses (CoVs) that affect severe illnesses in animals, including canine, feline, dromedary camels, porcine, bovine, and birds, are now identified for causing severe respiratory, gastrointestinal, neurological, and systemic diseases in diverse animal species. Significant initiatives are underway to identify potent therapeutic methods for addressing coronavirus (CoV) infections. Nonetheless, the advancement is hindered by a constrained grasp of virus-triggered host responses and pathogenesis. Hence, we suggest utilizing naturally secreted extracellular vesicles (EVs), particularly exosomes, to investigate host-pathogen responses after CoV infection.Exosomes leverage common endosomal sorting pathways and mechanisms, resembling the process seen in viruses. Exosomes play a significant role in cell-to-cell transmission of viral disease, and they function as cargo to carry information from donor cells to nearby and distant target cells. Simultaneously, exosomes can influence various cellular processes, including coagulation, inflammation, and immunomodulation, throughout SARS-CoV-2 infection. Therefore, we hypothesize that CoV hijacks the host exosomal pathway, thereby influencing the host EV biogenesis and composition within distinct host cellular models. This bears a resemblance to the mechanisms observed in other viral infections.Herein, Crandell-Rees Feline Kidney (CRFK) cells were infected with feline coronavirus (FCoV) in an exosome-free media at 0.005 multiplicity of infection (MOI), and A-72 fibrosarcoma cells were subjected to separate infections with feline and canine coronavirus (CCoV) in an exosome free media at 0.001 MOI, with incubation periods of 48h and 72h. The cell viability of CRFK and A-72 cells was significantly downregulated with increased incubation time, which was identified by performing the 3-(4,5-dimethylthiazo-1-2yl)-2,5- diphenyltetrazolium bromide (MTT) assay. After the infection, EVs were isolated through ultracentrifugation, and the subsequent analysis involved quantifying and characterizing the purified EVs using various techniques. The surface morphology of isolated EVs was analyzed via Scanning Electron Microscopy (SEM). NanoSight particle tracking analysis (NTA) verified that EV dimensions fell between 100 and 200 nm at both incubation periods. Total DNA, RNA, and protein levels were determined in CRFK-derived EVs at both incubation time points; however, total protein was significantly increased at 48h after FCoV infection. At both periods, total protein and RNA levels were significantly upregulated in A-72-derived EVs following FCoV and CCoV infections. However, total DNA levels were gradually upregulated with increased incubation time. Dot blot analysis indicated that the expression levels of angiotensin-converting enzyme 2 (ACE2), transmembrane protease, serine 2 (TMPRSS2), interleukin-1 beta (IL-β1), flotillin-1, ALG-2-interacting protein x (Alix), tumor susceptibility gene 101 (TSG101), cluster of differentiation (CDs) (29,47,63), toll-like receptors (TLRs) (3,6,7), tumor necrosis factor-alpha (TNF-α), caspases (1,8), and heat shock proteins (Hsps) were modified in CRFK and A72-derived EVs following CoV infections. Our finding indicated that FCoV and CCoV infections could regulate the EV formation and content, influencing pathogenicity and disease advancement. Investigating diverse animal CoVs will provide in-depth insight into host exosome biology during CoV infection and how EVs significantly influence viral pathogenesis via their function in viral-host interaction. Hence, this study provides valuable insights into CoV transmission and immune response regulation, and such understanding could provide information to prevent future CoV outbreaks.

English

ISBN: 9798304996518Subjects--Topical Terms:

536250
Microbiology.
Subjects--Index Terms:

Canine coronavirus

Comparative Assessment of Feline and Canine Coronavirus-Induced Modifications in Extracellular Vesicle Biogenesis and Composition in Host Cells /
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