Ritchie, Morgan
(2024)
Using a SARS-CoV-2 Pseudotyped Lentivirus to Determine the Tropism in Human Lung Cell Suspensions.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the novel, positive-sense RNA coronavirus was discovered in late December 2019 in Wuhan, China. This virus causes COVID-19 and was responsible for a global pandemic. There have been over 775 million cases reported worldwide, and over 7 million deaths worldwide. COVID-19 symptoms vary from person to person usually depending on age, sex, and pre-existing illnesses. Symptoms could range anywhere from a mild cold to severe pneumonia and death. The surface SARS-CoV-2 is covered with a structural spike glycoprotein, which binds to human angiotensin-converting enzyme 2 (ACE2) to infect susceptible cells. Most infection from SARS-CoV-2 takes place in epithelial cells in respiratory tract, with the main target being alveolar type II epithelial cells. While ACE2 is expressed on alveolar type II cells in the lungs, it is also expressed in the heart, kidneys, intestines, and other tissue.
Due to the SARS-CoV-2 spike protein’s ability to bind to ACE2, spike is a main target for vaccine development. However, new variants are constantly circulating, which can render current vaccines and treatments less effective. SARS-CoV-2 is currently a biosafety level three pathogen, so access to work with the virus is limited and comes with a certain level of risk associated with it. My project aims to develop a SARS-CoV-2 pseudotyped lentivirus that contains the spike glycoprotein from the Wuhan strain of SARS-CoV-2 and identify which cells in human lung cell suspensions are infected using microscopic fluorescent imaging and flow cytometry. We have observed that developing a pseudotyped lentivirus with the Wuhan SARS-CoV-2 spike protein is possible, but efficiency of the virus varies between cell lines and between different donors. Future steps will be needed to improve the efficiency of the pseudotyped virus.
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Details
| Item Type: |
University of Pittsburgh ETD
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| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
26 June 2024 |
| Date Type: |
Publication |
| Defense Date: |
10 June 2024 |
| Approval Date: |
26 June 2024 |
| Submission Date: |
21 June 2024 |
| Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
| Number of Pages: |
62 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Public Health > Infectious Diseases and Microbiology |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
SARS-CoV-2, pseudotyped lentivirus, human lung cell suspensions, Spike rotein |
| Date Deposited: |
26 Jun 2024 19:57 |
| Last Modified: |
26 Jun 2024 19:57 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/46608 |
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