Details

Autor(en) / Beteiligte

• Santos, Ana Catarina Taborda Godinho dos

Titel

Modulation of HIV-1-host cell pathways: From virus entry to latency

Ort / Verlag

ProQuest Dissertations & Theses

Erscheinungsjahr

2017

Quelle

ProQuest Dissertations & Theses A&I

Beschreibungen/Notizen

• AIDS represents a major global problem on human health due to increased number of HIV-1 resistant variants to antiretroviral drugs in infected individuals. HIV infection occurs following a series of virus-host interactions that not only lead to viral replication but also result in the generation of latently infected CD4+ T cells. Thus, targeting host cell factors involved in HIV-1 replication might be an alternative strategy to the currently available antiviral agents. Here we present strategies that allow the study of the function of specific cellular genes in HIV replication cycle through gene silencing. Using shRNA or siRNA constructs, we knockdown the expression of DNA-PKcs, EZH2, CIB2 and CIB1 in different cell lines and primary cells to verify the impact of the downmodulation of these candidates on HIV-1 replication. We observed a reduction of viral proteins expression and virus production, indicating for the first time that CIB1 is a helper factor for HIV-1, in addition to CIB2, EZH2 and DNA-PKcs. We also demonstrate that reduced expression of these proteins affected HIV-2 replication, suggesting them as HIV2 helper factors for the first time. Following this evaluation, we assessed the steps of the viral cycle that could be affected by these helper factors. EZH2 down-modulation did not impair HIV-1 replication in primary CD4+ T lymphocytes and facilitated viral reactivation in latently infected cell lines, suggesting that EZH2 is not indeed a helper factor for HIV-1. Both CIB1 and CIB2 act on early steps of the viral life cycle, facilitating HIV-1 entry in natural target cells in part through modulation of relevant cell surface molecules. Overall, this research work provides new insights for the complex host-HIV interactions instigating further studies that could raise new possibilities for antiviral strategies.