Characterization of compounds that inhibit human interferron-alfa activity using a new reporter gene assay

BURGI, MARÍA DE LOS MILAGROS; PRIETO, CLAUDIO; KRATJE, RICARDO; ETCHEVERRIGARAY, MARINA; OGGERO, MARCOS; BOLLATI-FOGOLÍN, MARIELA

Buenos Aires

Congreso; XLIX Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB).; 2013

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

Several pathologies and, in particular, systemic lupus erythematosus have shown an excessive production of endogenous IFN-alpha with undesirable symptoms for the organism. Because of that, it is interesting to find some molecules which can inhibit the activity of the above-mentioned cytokine. Following this objective, the WISH-Mx2/eGFP human reporter IFN activity cell line was obtained in our lab and employed to analyze 88 synthetic compounds gently transferred from Facultad de Ciencias, Universidad de la República, Montevideo, Uruguay. The reporter cell line has the Mx2/eGFP construction, where the sequence of enhanced green fluorescence protein (eGFP) is under the control of Mx2 promoter, specifically induced by type I human IFNs (hIFNs-I, among them, hIFN-alpha). Thus, the eGFP expression is directly correlated with the hIFN-I concentration in the sample. Using the reporter cell line, the eGFP expression in the presence and absence of each compound was compared aiming to find to some molecules that decrease the eGFP production as a consequence of IFNs-I activity inhibition. As a result, 10 compounds with the mentioned characteristics were identified. These compounds were extended characterized by studying their toxicity, effective dose, their effect on antiviral and antiproliferative IFNs-I activity, their action on cell cycle and the analysis of their apoptotic potentiality. Finally, 5 of them were selected to analyze their combined effect on the IFN biologic activity by employing the reporter gene assay. An interesting cooperative effect to decrease the IFN activity was observed. Besides, this effect was corroborated by the standard methods routinely used to study the IFN´s potency: antiviral and antiproliferative activity assay.