897 CONSENSUS RECOMMENDATIONS FOR VIROLOGIC NOMENCLATURE IN DAA CLINICAL TRIALS

Abstract

development, hippocampus and prefrontal cortex, through the use of primary cultured mouse neurons. Methods: Hippocampal and prefrontal cortex neurons were isolated from 18-day-old embryos of C57 BL/6 mice. After disaggregation with trypsin, they were cultured at 1.2×105 cells/well in DMEM and 10% FCS, and later in Neurobasal medium. At day 9 of culture, stimulation was performed using 100, 500 and 1000 IU of mouse IFN-a2a, and cells were collected after 12, 24 and 48 hours. The expression of interferon stimulated genes (ISGs) and 10 of the depression-related interferon-inducible genes (DRIIs) was assessed using real time PCR. Results: The stimulation with mouse IFN promoted an increased expression of the studied ISGs (Isg15, Mx2 and Ifit1) in a doseresponse manner, and, in general terms, they reached similar rates at the concentration and time points in both studied areas. Regarding to the DRIIs, differential responses to the treatment were observed, ranging from a nonor mild response in one or both studied areas (i.e. Dynlt1, Mef2A and Glrx showed a light increase in prefrontal cortex but not in hippocampus), to a strong upregulation, as found in Stat1, Rtp4, Mef2A and Ube2L6. Conclusions: IFN-a treatment promotes a response of ISGs in the cultured neurons of hippocampus and prefrontal cortex, and is also associated to the modulation of the expression of most of the studied DRIIs, meanwhile others might be under the regulation of factor(s) different than IFN-a. Linked to our previous findings, the upregulation of some DRIIs may be involved in the pathophysiological mechanisms underlying IFN-a therapyassociated depression.