High-throughput antibody-based assays to identify and quantify radiation-responsive protein biomarkers

Abstract

Purpose After a radiological ‘dirty bomb’ incident in a major metropolitan center, substantial numbers of people may be exposed to radiation. However, only a fraction of those individuals will need urgent medical attention. Consequently, a rapid screening test is needed to identify those people who require immediate treatment. Material and methods Ten normal human cell lines were screened by enzyme-linked immunosorbent assay (ELISA) for the expression of a dozen secreted cytokines that have been reported to have changes in protein or mRNA levels at 1, 2, and 3 days after 0–10 Gy irradiation using 137Cs gamma rays at 0.82 Gy min−1. After this systematic in vitro screen, we measured changes in the level of a subset of these candidate proteins in plasma from irradiated C57BL/6 mice (n = 3 per group), comparing shams with a single radiation dose (5 Gy X-rays) at 3.7 Gy min−1 at 6 h after irradiation. Results We identified four cytokine molecules that had altered levels after radiation exposure, one of which, Interleukin (IL) 6, was consistently elevated after irradiation in vitro and in vivo. Conclusions Our findings underscore the potential for IL6 as a marker for an immunoassay-based, rapid, high-throughput biodosimeter.

Exposure to ionizing radiation reveals global dose- and time-dependent changes in the urinary metabolome of rat

“…In this study, we expand upon earlier published observations of that have reported on the urinary and serum response of the mouse (Amendola et al 2006; Lee et al 2007; Tyburski et al 2008; 2009; Ossetrova et al 2010; Partridge et al 2010; Chen et al 2011) and rat(Randic & Supek 1961; Porciani et al 2001; Lanz et al 2009; Johnson et al 2011; Tang et al 2013; Zhang et al 2014) to γ radiation exposures. Using Ultra-Performance ® liquid chromatography (UPLC) in conjunction with time-of-flight mass spectrometry (TOFMS), we analyzed urine collected from rats prior to and at four time points after exposure to six different doses of gamma radiation ranging from 0.5 to 10 Gy at high dose rate.…”

Radiation-induced non-targeted response in vivo: role of the TGFβ-TGFBR1-COX-2 signalling pathway

“…Following 1 h incubation at 20 °C, microwells were washed four times and 100  μ l of tetramethylbenzidine (TMB, Thermo Scientific, Rockford, IL, USA) added for 15–30 min at 20 °C. Reactions were stopped by addition of 0.9 ℳ H 2 SO 4 (50  μ l) and product absorbance was determined at 450 nm (Partridge et al , 2010). …”

Oxidative DNA damage caused by inflammation may link to stress-induced non-targeted effects

“…Other cytokines shown to have potential influence on non-targeted effects include IL-6, Il-8, CCL2, CCL4, CCL7, CXCL7 and TNFα [12; 71; 92; 93]. Hei et al .…”