The increase in temperatures is promoting the spread of ticks and the viruses they transmit even in areas previously unaffected. Ticks, characterized by a long life cycle and by mechanisms such as trans-stadial and trans-ovarial transmission, can carry arboviruses in a persistent manner, and viral replication can be influenced by environmental temperature. In this work, the effect of temperature on the replication of Hazara virus (HAZV), a study model for Crimean-Congo hemorrhagic fever virus, was analyzed in HAE/CTVM8 tick cells. The data show that 28–32 °C represent favorable conditions for cell survival and proliferation and for the persistence of infection, whereas at 37 °C marked stress and reduced viability were observed. Cells stably adapted to 32 °C displayed more robust growth and greater tolerance to infection compared to those maintained at 28 °C, suggesting a key role of physiological adaptation. The comparison between immunofluorescence and RT-qPCR highlighted different but complementary trends: RT-qPCR detected higher values and more pronounced dynamics, whereas immunofluorescence returned lower values and flatter curves, respectively reflecting intracellular replication of the viral genome and the production of extracellular infectious particles. Overall, the results confirm the importance of temperature as a determining variable in the infection dynamics of HAZV, highlighting both methodological limitations and the potential of different experimental approaches. In perspective, more gradual adaptation protocols, comparison with other cell lines, and integration with ecological and climatic data could improve data reliability. Indeed, linking in vitro experimental observations with real environmental scenarios will allow the translation of results into useful insights for the understanding and management of arboviruses in the context of climate change.
L’incremento delle temperature sta promuovendo la diffusione delle zecche e dei virus da esse trasmessi anche in aree precedentemente non interessate. Le zecche, caratterizzate da un ciclo vitale lungo e da meccanismi come la trasmissione trans-stadiale e trans-ovarica, possono veicolare gli arbovirus in maniera persistente, e la replicazione virale può essere influenzata dalla temperatura ambientale. In questo lavoro è stata analizzata l’influenza della temperatura sulla replicazione del virus Hazara (HAZV), modello di studio del virus della febbre emorragica della Crimea e Congo, in cellule di zecca HAE/CTVM8. I dati mostrano che 28–32 °C rappresentano condizioni favorevoli per la sopravvivenza e la proliferazione delle cellule e per la persistenza dell’infezione, mentre a 37 °C si osservano stress marcato e ridotta vitalità. Le cellule adattate stabilmente a 32 °C hanno mostrato una crescita più robusta e una maggiore tolleranza all’infezione rispetto a quelle mantenute a 28 °C, suggerendo un ruolo chiave dell’adattamento fisiologico. Il confronto tra immunofluorescenza e RT-qPCR ha evidenziato andamenti diversi ma complementari: la RT-qPCR ha rilevato valori più elevati e dinamiche più marcate, mentre l’immunofluorescenza ha restituito valori inferiori e curve più piatte, riflettendo rispettivamente la replicazione intracellulare del genoma virale e la produzione di particelle infettive extracellulari. Nel complesso, i risultati confermano l’importanza della temperatura come variabile determinante nella dinamica di infezione di HAZV, mettendo in luce limiti metodologici e potenzialità dei diversi approcci sperimentali. In prospettiva, protocolli di adattamento più graduali, il confronto con altre linee cellulari e l’integrazione con dati ecologici e climatici potrebbero migliorare l’affidabilità dei dati. Collegare le osservazioni sperimentali in vitro con scenari ambientali reali permetterà infatti di tradurre i risultati in indicazioni utili per la comprensione e la gestione degli arbovirus in un contesto di cambiamento climatico.
Valutazione degli effetti della temperatura sulla replicazione dei nairovirus nelle cellule di zecca
BENETTON, CECILIA
2024/2025
Abstract
The increase in temperatures is promoting the spread of ticks and the viruses they transmit even in areas previously unaffected. Ticks, characterized by a long life cycle and by mechanisms such as trans-stadial and trans-ovarial transmission, can carry arboviruses in a persistent manner, and viral replication can be influenced by environmental temperature. In this work, the effect of temperature on the replication of Hazara virus (HAZV), a study model for Crimean-Congo hemorrhagic fever virus, was analyzed in HAE/CTVM8 tick cells. The data show that 28–32 °C represent favorable conditions for cell survival and proliferation and for the persistence of infection, whereas at 37 °C marked stress and reduced viability were observed. Cells stably adapted to 32 °C displayed more robust growth and greater tolerance to infection compared to those maintained at 28 °C, suggesting a key role of physiological adaptation. The comparison between immunofluorescence and RT-qPCR highlighted different but complementary trends: RT-qPCR detected higher values and more pronounced dynamics, whereas immunofluorescence returned lower values and flatter curves, respectively reflecting intracellular replication of the viral genome and the production of extracellular infectious particles. Overall, the results confirm the importance of temperature as a determining variable in the infection dynamics of HAZV, highlighting both methodological limitations and the potential of different experimental approaches. In perspective, more gradual adaptation protocols, comparison with other cell lines, and integration with ecological and climatic data could improve data reliability. Indeed, linking in vitro experimental observations with real environmental scenarios will allow the translation of results into useful insights for the understanding and management of arboviruses in the context of climate change.| File | Dimensione | Formato | |
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https://hdl.handle.net/20.500.12608/92054