Type I IFNs: new reporter cell lines to determine their biological activity

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

Potrero de los Funes

Congreso; XLVII Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular (SAIB); 2011

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

The potency of interferons (IFN) must be correctly identified because they are used as biopharmaceuticals since they have an important role in the immune defence. For this purpose, 4 new human reporter cell lines derived from different tissues (A549, Hela, Wish and HEp-2) were developed using the eGFP gene under the control of type I IFN-inducible Mx2 promoter. The Mx2 promoter responded specifically and quantitatively to type I IFNs (IFN-alfa and -beta) showing a good correlation between the eGFP expression and the IFNs added. Comparing the cell lines responsiveness to IFN-alfa subtypes, no differences in detection limit were observed among them. Contrarily, differences were observed when comparing the activity of IFN-beta subtypes in HEp-2 cell lines. Therefore, the standardization of several assays to measure the potency of IFNs might be carried out using the cell lines herein documented. These systems have many advantages with respect to already existing assays: they are very fast, they can determine the potency of type I IFNs using only one cell line and have the specificity of the Mx promoter. They are sensitive and safe, showing reproducible responses in a dose dependent manner. Outstandingly, the main contribution of this work was the development of alternative reporter systems as suitable candidates to evaluate the way that IFNs induce their activity in different human tissues.