Searching for compounds that modulate type I interferon activity by screening libraries with new reporter cell lines

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

Lille

Congreso; 23rd ESACT Meeting; 2013

Le Public Système PCO - European Society for Animal Cell Technology

IFNs have dual clinical effects depending on their use as pharmaceuticals (IFN-alpha or IFN-beta) or their special feature of being self-produced by humans in some autoimmune diseases (IFN-alpha). Thus, increasing their therapeutic efficiency in the first situation or decreasing the side effects in the second one involve high clinical value. In this sense, it is interesting to find molecules which can modulate the IFN´s activity. We carried out a simple, fast and robust reporter assay to identify IFN activity modulator compounds. We analyzed lots of them simultaneously giving reliable and reproducible results employing Mx2/eGFP modified-A549, -HeLa, -HEp2 and -WISH reporter cell lines previously developed in our lab. In these cell lines the enhanced green fluorescence protein (eGFP) gene is driven by the specific type I IFNs inducible Mx2 promoter. The eGFP percentage obtained after type I IFN addition is directly correlated with the cytokine in the sample. We used the reporter cell lines to analyze a complete natural library of 176 compounds and 288 from a synthetic library of 2,500 compounds provided by the Helmholtz Zentrum für Infektionsforschung, Braunschweig, Germany. We isolated 21 inhibitory compound and 1 enhancer compound. 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. Along this screening, we could easily discriminate positive from negative compounds. The Z factor value, commonly used in high throughput screening, 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.