INVESTIGADORES
ATTALLAH Carolina Veronica
congresos y reuniones científicas
Título:
Characterization of an anti-interferon-alpha2b chimeric fusion protein for systemic lupus erythematosus therapeutic use
Autor/es:
ATTALLAH C; MUFARREGE, E; ETCHEVERRIGARAY, M; OGGERO, M
Lugar:
Buenos Aires
Reunión:
Congreso; IV International Congress in Translational Medicine; 2018
Institución organizadora:
Universidad de Buenos Aires
Resumen:
Systemic lupus erythematosus (SLE) is a chronic, multisystem, autoimmune disease that predominantly affects women of childbearing age. Its clinical manifestations (arthritis, rashes, alopecia, vasculitis, nephritis, serositis) lead to significant morbidity, reduced physical function, loss of employment, impaired quality of life, high risk of permanent disability and shortened life span. Treatment of SLE remains challenging because of the suboptimal efficacy of standard-of-care medications and the serious adverse events associated with their use. Since the late 70?s, the interferon alpha (IFN alpha) has been associated with SLE. From that moment, a remarkable number of studies have confirmed the importance of IFN-alpha in SLE, and have prompted the investigation of anti IFN alpha therapeutic strategies. Indeed, several clinical trials have been initiated using monoclonal antibodies (mAb) against IFN-alpha.Our group has generated a chimeric anti-IFN-alpha2b fusion protein as a strategy to develop a biotherapeutic for SLE in Argentine. This chimeric protein comprises a single chain variable fragment (scFv) derived from a murine mAb anti-IFN-alpha2b and the Fc gamma1 region from human antibodies. In this work, we have characterized the anti-IFN-alpha2b chimeric protein produced in different types of mammalian cells by analyzing IFN-alpha affinity, in vitro biological activity and the ability to produce antibodies in transgenic animals. CHO-K1 (Chinese hamster ovary), HEK293 (Human Embryonic Kidney) and NS0 (derived from murine myeloma) cells were used. The chimeric proteins produced by each cell line demonstrated affinities in the order of 10-8 M-1 and the ability to neutralize the IFN-alpha antiproliferative and antiviral activities. The best performance was obtained by the CHO K1-produced chimeric protein. On the other hand, each chimeric protein showed different thermal stability depending on the producing host. In addition, HLA-DR1 transgenic mice inoculated with the distinct fusion proteins generated different humoral immune responses in terms of antibody titles. The highest antibody title was obtained by inoculating the HEK293 cell-derived protein. This characterization allows us to take into account the importance to select the proper host in order to project an anti-IFN-alpha2b antibody as a potential therapeutic agent for SLE.