Crimean-Congo haemorrhagic fever (CCHF) is the most common tick-borne disease, it has a mortality rate of 30% to 70% and is caused by Crimean-Congo haemorrhagic fever virus (CCHFV). Considering the high pathogenicity of CCHFV, handling the virus requires the use of biosafety level 4 (BSL-4) laboratories. Its pathogenesis remains poorly understood and no specific therapy has yet been identified. The virus nucleoprotein (NP) seems to play a role in the pathogenesis of the disease, with a mechanism that has not yet been clarified. This thesis is part of a main project aimed at investigating the role of NP in viral pathogenesis by assessing the induction of inflammatory cytokines, which are a marker of viral infection, and identifying possible cellular interactors that may interact with the nucleoprotein. To this end, the two NP variants derived from the CCHFV IbAr 10200 strain, a laboratory-adapted strain considered non-pathogenic, and the highly pathogenic Hoti-WT strain were selected. As a control, NP from Hazara virus, a non-pathogenic virus considered to be a good surrogate for CCHFV, was also included. The aim of the thesis is to develop an efficient transduction protocol, with the ultimate goal of using the protocol in target cells of viral infection in which the different nucleoprotein variants can be expressed and thus assessing their effect on the cell. In particular, the efficiency of three transduction protocols was evaluated in the thesis work, in order to select the method that guarantees the highest levels of NP expression.
La febbre emorragica della Crimea-Congo (CCHF) è la più diffusa malattia trasmessa dalle zecche, ha un tasso di mortalità che varia dal 30% al 70% ed è causata dal virus della febbre emorragica della Crimea-Congo (CCHFV). Considerando l'elevata patogenicità del CCHFV, la manipolazione del virus richiede l'uso di laboratori di biosicurezza di livello 4 (BSL-4). La sua patogenesi rimane ancora poco conosciuta e non è ancora stata individuata una terapia specifica. La nucleoproteina (NP) del virus sembra svolgere un ruolo nella patogenesi della malattia, con un meccanismo non ancora chiarito. Questa tesi si inserisce in un progetto più ampio volto a investigare il ruolo della NP nella patogenesi virale attraverso la valutazione dell’induzione di citochine infiammatorie, che rappresentano un marker di infezione virale, e l’identificazione di eventuali interattori cellulari che possono interagire con la nucleoproteina. A tal fine, sono state selezionate le due varianti di NP derivanti dal ceppo CCHFV IbAr 10200, un ceppo adattato in laboratorio considerato non patogeno, e il ceppo altamente patogeno Hoti-WT. Come controllo, è stata anche inclusa la NP del virus Hazara, un virus non patogeno considerato essere un buon surrogato del CCHFV. Lo scopo della tesi è sviluppare un efficiente protocollo di trasduzione, con l’obbiettivo finale di utilizzare il protocollo in cellule bersaglio dell’infezione virale in cui far esprimere le differenti varianti di nucleoproteina e valutarne quindi l’effetto sulla cellula. In particolare, nel lavoro di tesi è stata valutata l’efficienza di tre protocolli di trasduzione, per selezionare il metodo che garantisca i maggiori livelli di espressione della NP.
Valutazione dell'azione patogenetica della nucleoproteina del virus della febbre emorragica della Crimea-Congo
REOLON, ARIANNA
2021/2022
Abstract
Crimean-Congo haemorrhagic fever (CCHF) is the most common tick-borne disease, it has a mortality rate of 30% to 70% and is caused by Crimean-Congo haemorrhagic fever virus (CCHFV). Considering the high pathogenicity of CCHFV, handling the virus requires the use of biosafety level 4 (BSL-4) laboratories. Its pathogenesis remains poorly understood and no specific therapy has yet been identified. The virus nucleoprotein (NP) seems to play a role in the pathogenesis of the disease, with a mechanism that has not yet been clarified. This thesis is part of a main project aimed at investigating the role of NP in viral pathogenesis by assessing the induction of inflammatory cytokines, which are a marker of viral infection, and identifying possible cellular interactors that may interact with the nucleoprotein. To this end, the two NP variants derived from the CCHFV IbAr 10200 strain, a laboratory-adapted strain considered non-pathogenic, and the highly pathogenic Hoti-WT strain were selected. As a control, NP from Hazara virus, a non-pathogenic virus considered to be a good surrogate for CCHFV, was also included. The aim of the thesis is to develop an efficient transduction protocol, with the ultimate goal of using the protocol in target cells of viral infection in which the different nucleoprotein variants can be expressed and thus assessing their effect on the cell. In particular, the efficiency of three transduction protocols was evaluated in the thesis work, in order to select the method that guarantees the highest levels of NP expression.File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/34828