Production of HIV-1 polypeptides

Dissertação de Erasmus Mundus para obtenção do grau de mestre em Técnicas Laboratoriais ForensesHIV causes acquired immunodeficiency syndrome known as AIDS. HIV-1 and HIV-2 are the etiologic agents of AIDS, and despite very similar structural and genomic organization, HIV-1 and HIV-2 infections are very different regarding immunological and clinical outcomes. Neutralizing antibody responses, both autologous and heterologous is more frequent in HIV-2 than in HIV-1 infection. HIV-1 can escape naturally occurring antibodies (Nabs) due to sequence variability and epitope masking by glycosylation, which can cause difficulties and challenges in vaccine development. Even though is still debatable, region in HIV-2 that may contain broadly neutralizing epitotes is the V3 region. Furthermore, in HIV-2 envelope complex C2 and C3 regions are unprotected and therefore under strong diversifying selection, which can indicate that, like in HIV-1, they may harbor neutralizing epitopes. The hypothesis beneath this project is that an chimeric envelope surface glycoprotein containing the C2, V3 and C3 regions of HIV-2 and the remaining regions of env of recombinant AE subtype HIV-1 92TH019 might elicit a broadly neutralizing response against both HIV-1 and HIV-2. Aim of this project is the production of HIV-1 polypeptides (V1V2, C5, C3V4C4, V4) derived from recombinant AE subtype HIV-1 92TH019 which will be further used in immunization boosting in mice with vaccine, as well as detection of antibodies in mice. In this study these regions were successfully amplified and purified. Annealing temperatures that gave best amplification results were 49°C (C3V4C4 region), 50°C (V1V2 region), 50°C (C5 region) and 49°C (V4 region). To my best knowledge this is first time that these regions were amplified from this subtype. Due to the lack of time, cloning and expression of these regions was not possible, however research will be continued