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Specific elimination of HIV-1 infected cells using Tat/Rev-dependent circuit

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Detalhes bibliográficos
Resumo:Despite the success of antiretroviral cocktails, a cure for HIV-1 remains elusive. This is mainly due to the existence of persistent cellular reservoirs infected with non-transcriptional, latent HIV-1. An effective treatment against HIV-1 would target both active and latent HIV-1-infected cells, and eliminate them without harming non-infected cells. In order to achieve this, we have developed a therapeutic plasmid with a Tat/Rev-dependent toxin expression. This vector contains a HIV-1 transcriptional activator to specifically activate latent infected cells. We confirmed the expression of all vector components necessary to achieve our goal. We then assessed the potential of this therapeutic plasmid to specifically eliminate infected cells by transduction of a T-lymphoma cell line that has a HIV-1 latent provirus integrated in its genome. We demonstrated that our therapeutic plasmid is not only capable of reactivating latent HIV-1 expression by itself, but it also can induce cell death efficiently and specifically in HIV-1 infected cells. Cell death was 60% higher in infected cells than in non-infected cells, indicating that our therapeutic vector was able to fulfill the objective proposed in this work. Therefore, the data presented here represents a promising approach for the development of a gene therapy that could become a viable and successful treatment against HIV-1.
Autores principais:Perdigão, Pedro Ricardo Lucas, 1987-
Assunto:HIV-1 Terapia génica Retrovírus Teses de mestrado - 2011
Ano:2011
País:Portugal
Tipo de documento:dissertação de mestrado
Tipo de acesso:acesso aberto
Instituição associada:Universidade de Lisboa
Idioma:inglês
Origem:Repositório da Universidade de Lisboa
Descrição
Resumo:Despite the success of antiretroviral cocktails, a cure for HIV-1 remains elusive. This is mainly due to the existence of persistent cellular reservoirs infected with non-transcriptional, latent HIV-1. An effective treatment against HIV-1 would target both active and latent HIV-1-infected cells, and eliminate them without harming non-infected cells. In order to achieve this, we have developed a therapeutic plasmid with a Tat/Rev-dependent toxin expression. This vector contains a HIV-1 transcriptional activator to specifically activate latent infected cells. We confirmed the expression of all vector components necessary to achieve our goal. We then assessed the potential of this therapeutic plasmid to specifically eliminate infected cells by transduction of a T-lymphoma cell line that has a HIV-1 latent provirus integrated in its genome. We demonstrated that our therapeutic plasmid is not only capable of reactivating latent HIV-1 expression by itself, but it also can induce cell death efficiently and specifically in HIV-1 infected cells. Cell death was 60% higher in infected cells than in non-infected cells, indicating that our therapeutic vector was able to fulfill the objective proposed in this work. Therefore, the data presented here represents a promising approach for the development of a gene therapy that could become a viable and successful treatment against HIV-1.