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INVESTIGATION OF CYTOKINE PRODUCTION IN MACROPHAGES IN INTERACTION WITH SARS-COV-2 virus RECEPTOR BINDING DOMAIN
Malmö University, Faculty of Health and Society (HS).
2022 (English)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
UNDERSÖKNING AV Cytokinproduktion HOS makrofager vid INTERAKTION med SARS-CoV-2 virusets RECEPTORBINDANDE DOMÄN (Swedish)
Abstract [en]

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the virus that causes coronavirus disease-2019 (COVID-19). The virus was first identified in December 2019, spread worldwide, and caused many deaths. The World Health Organization (WHO) declared a global public health crisis on 30 January 2020 and a pandemic on 11 March 2020. SARS-CoV-2 is a single-stranded RNA virus belonging to the genus Coronavirus and the family Coronaviridae. SARS-CoV-2 has four structural proteins, Envelope (E), Membrane (M) and Nucleocapsid (N) and Spike (S) proteins. The S protein consists of S1 and S2 subunits. The receptor-binding domain (RBD) of the virus is located in the S1 subunit and binds the virus to the surface receptor angiotensin convertase-2 (ACE2) of the host cell. This study aimed to better understand the production of proinflammatory cytokines in macrophages after in vitro interaction with the RBD of SARS-CoV-2. Gene expression of tumor necrosis factor alpha (TNF-α), interleukin 6 (IL-6) and interleukin-1 Beta (IL-1β) in RBD stimulated mouse macrophage cell line RAW 264.7 was detected with quantitative Reverse Transcription Polymerase Chain Reaction (RT-qPCR). In addition, the concentration of secreted TNF-α, IL-6 and IL-1β was quantified with sandwich enzyme-linked immunosorbent assay (ELISA). The study indicated that mouse macrophages produce TNF-α upon exposure to SARS-CoV-2 RBD after 6 and 24 hours of cell stimulation, but neither IL-6 nor IL-1β were increased after stimulation. The future goals of the study are to understand how the immune system responds to the SARS-CoV-2 infection, and how this can help in treating COVID-19.

Abstract [sv]

Allvarligt akut respiratoriskt syndrom coronavirus-2 (SARS-CoV-2) är det virus som orsakar coronavirussjukdom-2019 (COVID-19). Viruset identifierades först i december 2019 och spreds sedan över hela världen och orsakade många dödsfall. Världshälsoorganisationen (WHO) utropade en global folkhälsokris den 30 januari 2020 och en pandemi den 11 mars 2020. SARS-CoV-2 är ett enkelsträngat RNA-virus som tillhör släktet Coronavirus och familjen Coronaviridae. SARS-CoV-2 har fyra strukturella proteiner, hölje (E), membran (M) och nukleokapsid (N) och Spike (S) proteiner. S-proteinet består av S1- och S2-subenheter. Den receptorbindande domänen (RBD) av viruset är belägen i S1-subenheten och binder viruset till ytreceptorn angiotensinconvertas-2 (ACE2) i värdcellen. Denna studie syftade till att bättre förstå produktionen av proinflammatoriska cytokiner i makrofager efter in vitro-interaktion med RBD av SARS-CoV-2. Genuttryck av tumörnekrosfaktor alfa (TNF-α), interleukin 6 (IL-6) och interleukin-1 Beta (IL-1β) i RBD-stimulerad musmakrofagcellinje RAW 264.7 detekterades med kvantitativ Reverse Transcription Polymerase Chain Reaction (RT-qPCR). Dessutom kvantifierades koncentrationen av utsöndrad TNF-α, IL-6 och IL-1β med sandwich enzyme-linked immunosorbent assay (ELISA). Studien visade att musmakrofager producerar TNF-α vid exponering för SARS-CoV-2 RBD efter 6 och 24 timmars cellstimulering, men varken IL-6 eller IL-1β ökade efter stimuleringen. De framtida målen för studien är att förstå hur immunsystemet svarar på SARS-CoV-2-infektionen och hur detta kan vara till hjälp för att behandla COVID-19.

Place, publisher, year, edition, pages
2022. , p. 21
Keywords [en]
ACE2, Covid-19, IL-1β, IL-6, Macrophages, RBD, SARS-CoV-2, S-protein, TNF-α
Keywords [sv]
ACE2, Covid-19, IL-1β, IL-6, Makrofager, RBD, SARS-CoV-2, S-protein, TNF-α
National Category
Biomedical Laboratory Science/Technology
Identifiers
URN: urn:nbn:se:mau:diva-52258OAI: oai:DiVA.org:mau-52258DiVA, id: diva2:1665749
Educational program
HS Biomedicinsk laboratorievetenskap
Supervisors
Examiners
Available from: 2022-06-08 Created: 2022-06-08 Last updated: 2022-06-08Bibliographically approved

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