Endotoxin Response Profiling in Healthy Individuals

The GENE Study recruited healthy individuals (N = 294, 52% Female, 65% EA, 35% AA) ancestry to a University of Pennsylvania (UPenn) inpatient Clinical and Translational Research Center (CTRC) protocol as previously described (clinicaltrials.gov NCT00953667).[30 (link)] Participants completed an endotoxin challenge (1ng/kg E coli-derived LPS; U.S. standard reference, lot No. CCRE-LOT-1+2, Clinical Center, Pharmacy Department at the National Institutes of Health, Bethesda MD).[30 (link), 31 (link)] Multiple clinical variables were assessed during the visit. Individuals were ranked by their peak clinical inflammatory response (Δ from baseline for fever, plasma TNFα, plasma IL-6). Individuals falling within the top and bottom 5% of responses were designated as “high” or “low” responders respectively. CD14+ monocytes were isolated from whole blood using magnetic bead selection (Dynabeads^® CD14, ThermoFisher, Waltham MA), and frozen in TRIzol reagent (ThermoFisher, Waltham MA) for subsequent RNA extraction via standard protocol.[32 ]. The GENE study was approved by UPenn’s Institutional Review Board (IRB), with regulatory oversight by the FDA (LPS: IND# 5984) and an NIH-appointed data-safety and monitoring board. All subjects provided written informed consent.