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Vpliv strukturnih proteinov M, N, S in E koronavirusa SARS-CoV-2 na izločanje citokinov in izražanje proteinov APOBEC3 v človeških celicah A549 in HuH-7 = The effect of M, N, S, and E structural proteins of the SARS-CoV-2 virus on cytokine secretion and expression of APOBEC3 proteins in human A549 and HuH-7 cells : enovit magistrski študijski program FarmacijeTurk, Špela, farmacevtkaPandemija koronavirusne bolezni 2019 (COVID-19), ki jo povzroča hudi akutni respiratorni sindrom koronavirus-2 (SARS-CoV-2), je močno vplivala na zdravje ljudi po vsem svetu. Visoka prenosljivost in ... patogenost virusa je v zadnjih letih vzpodbudila znaten napredek v raziskavah, vključno z velikimi preboji v razvoju cepiv, možnostih zdravljenja, razumevanju prenosa in pojavljanja novih različic. Virus SARS-CoV-2 je RNA virus iz družine Betacoronavirusov, sestavljen iz strukturnih proteinov M, N, S, E, nestrukturnih in pomožnih proteinov. V gostiteljske celice vstopi prek encima angiotenzin konvertaza 2 in se v njih začne razmnoževati, celice gostitelja pa so opremljene z mehanizmi, ki jim omogočajo hitro prepoznavanje virusov in odziv nanje. Mednje sodijo tudi citokini in katalitičnemu polipeptidu podobni encimi, ki preurejajo mRNA apolipoproteina B (APOBEC), ki se začnejo v sklopu imunskega odziva ob okužbi pospešeno izražati in sodelovati pri inaktivaciji virusa. V magistrskem delu smo želeli raziskati, kako prekomerna izraženost strukturnih proteinov M, N, S, E SARS-CoV-2 vpliva na izražanje citokinov in proteinov APOBEC3 v celičnih linijah A549 in Huh-7. Ustrezno pripravljene celice smo najprej transficirali s plazmidi z zapisi za SARS-CoV-2 strukturne, pomožne in kontrolne proteine. Po 48 urah smo z encimskoimunskim testom preverili prisotnost citokinov IL-6 v celičnem gojišču. Iz transficiranih celic smo izolirali mRNA, jo s pomočjo reverzne transkripcije prepisali v stabilnejšo komplementarno DNA in na koncu pridobili kvantitativne rezultate izražanja genov za posamezne citokine in proteine APOBEC3 z verižno reakcijo s polimerazo v realnem času. Ugotovili smo, da v celicah Huh-7 izražanje mRNA citokinov IL-8 in TNF-α ter proteinov A3B in A3F najbolj poviša vnos strukturnih proteinov S, E in M. Vpliv pomožnega proteina ORF6 pri celicah A549 kaže na sorazmerje med izražanjem citokinov in proteinov iz družine APOBEC, raziskava ELISA pa ni pokazala povečanega izločanja citokina IL-6 v supernatantih preiskovanih celičnih vzorcih. Iz naših rezultatov sklepamo, da strukturni proteini S, E in M vplivajo na povišano izražanje citokinov in proteinov APOBEC in izzovejo imunski odziv.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [Š. Turk], 2024Language - slovenianCOBISS.SI-ID - 195577091
Author
Turk, Špela, farmacevtka
Other authors
Lovšin, Nika
Topics
SARS-CoV-2 |
Citokini |
proteini APOBEC3 |
encimskoimunski test |
A549 |
Huh-7 |
transfekcija
Library/institution |
City | Acronym | For loan | Other holdings |
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Faculty of Pharmacy, Ljubljana | Ljubljana | FFALJ |
reading room 1 cop.
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Turk, Špela, farmacevtka | |
Lovšin, Nika | 19649 |
Source: Personal bibliographies
and: SICRIS
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