Exploration of natural and synthetic libraries compounds by using new biological tools to detect some molecules which can modulate the Interferon type I activity

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

Puerto Varas

Congreso; XII Congreso de la Asociación Panamericana de Bioquímica y Biología Molecular (PABMB); 2013

Asociación Panamericana de Bioquímica y Biología Molecular (PABMB)

The clinical effects of IFNs depend on their use as biotherapeutics or their self-production in some autoimmune diseases. Therefore, the development of new IFN-I reporter assays could be of a high clinical value in order to search for compounds that could increase the IFNs therapeutic efficiency or decrease their side effect in the mentioned illnesses. With the aforementioned aim in mind, simple, fast and robust reporter assays were developed to analyze lots of compounds simultaneously giving reliable and reproducible results. Four human tissue-derived cell lines: WISH (derived via HeLa contaminantion), A549 (lung cancer cells), HEp-2 (epidermoid larynx carcinoma cells) and HeLa (cervical adenocarcinoma cells) were used to develop new reporter gene assays based on the expression of the eGFP gene under the control of type I IFN-inducible Mx2 promoter. Thus, the eGFP expression was directly correlated with the hIFN-I concentration in the sample. We analyzed a natural library of 176 compounds and 288 from a synthetic library provided by the Helmholtz Zentrum für Infektionsforschung, Braunschweig, Germany. We isolated 21 inhibitory and 1 enhancer compounds. All of them were characterized according to their cytotoxicity, effective doses, antiviral and anti-proliferative activity, residual and reversible effect, and their action on cell cycle and apoptosis. We could easily discriminate positive from negative compounds. The Z factor, was always higher than 0.8, reflecting the excellent quality of the bioassay. This search for compounds which can improve or block type I IFN´s activity shows a big potential in view of their therapeutic implications. We are very grateful to Mario Köster, Hansjörg Hauser, Dieter Kaufmann and Florenz Sasse for being so kind in providing us the compounds libraries and for the exhaustive help during this work.