Cloned of Sag in Virus like particles of the Junin Z protein (Z-VLP) as potential biological vehicle therapeutic tool.

SCARFO MARINA; PASTORINI MERCEDES; DE GANZÓ AGUSTÍN; BORIO CRISTINA; GOÑI SANDRA; ALEMÁN MERCEDES; DUARTE ALEJANDRA

Mar del Plata

Congreso; Reunión Anual de Sociedades de Biociencia - SAIC; 2019

SAIC

Superantigens are proteins that bind to molecules of the major histocompatibility class II complex and then interact with a specific Vβ chains in T cell receptors. We have previously described that Sags are able to induce apoptosis of murine lymphoma cells in vitro and in vivo. Besides, the advance in genetic engineering allowed the generation of virus-like particles (VLP), these molecules maintain the same structural properties of virions but without genome, so they are not infective. These constructions are considered very efficient as platforms for vaccines and transport systems of potentially therapeutic molecules. In this study we propose to use this property to obtain a platform with VLPs carrying Sag Vβ14 fused to Z. Arenavirus matrix protein Z plays an important role in virus budding and is able to generate enveloped Z-VLP in absence of any other viral proteins. We previously demonstrated that Z-VLP induce maturation of dendritic cells (DCs) derived from bone marrow Balb/c.In different species, including man, it has been observed that many T lymphomas have a mono or oligoclonal character, in terms of the expression of a given Vβ region. To that, VLP containing JUNV Z protein to the antigen presentation design based. First we study the capacity of the sequences of the Sag to interact with and specific TCR Vβ14 region. The sequence was amplified and cloned in the Z vector using the NotI and BamHI restriction sites. Four positive clones were sequenced and analyzed by MEGA Software. This positive plasmid was transfected on 293T mammalian cells, and VLPs were purified from the supernatant. Then we determinate the effect of this construct on reactive lymphocytes analyzing the induction of apoptosis by annexin V. The advantage of this plataform is the generation of new tool to direction the Sag expression on antigen pesentations cells to induce apoptosis of specific T-cell.