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

SCARFO MARINA; CRISTINA SILVIA BORIO; DUARTE ALEJANDRA; PASTORINI MERCEDES; SANDRA GOÑI; AGUSTÍN DE GANZÓ; ALEMAN MECEDES

Mar del Plata, Buenos Aires, Argentina

Conferencia; Reunión conjunta de Sociedades de Biociencias; 2019

SAIC-SAFE

In different species including man, many T lymphomas have a mono or oligoclonal character, in terms of the expression of a given Vβ region. Superantigens (Sags) are proteins that bind to molecules of the major histocompatibility class II complex (MHC II) and 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), which maintain the same structural properties of virions without genome so, they are not infective. In this study we designed a VLP platform carrying Sag Vβ14 fused to JUNV Z protein, as these constructions are very efficient platforms for vaccines and transport systems of therapeutic molecules. In this way, arenavirus matrix protein Z plays an important role in virus budding allowing generating enveloped Z-VLP in absence of any other viral proteins. First, we observed that Z-VLP induces maturation of bone marrow-derived dendritic cells from Balb/c which is a prerequisite for Sag-TCR interaction. Then we determined the interaction between Sag sequences and the specific Vβ14 region in TCR. Finally, 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. Thereafter, positive plasmid was transfected on 293T mammalian cells, and VLPs were purified from the supernatant. Next step will be to evaluate the capability to induce apoptosis of reactive lymphocytes by the construct to be considered as a new tool to eliminate specific T-cell.