We have developed a simple and reliable method to measure the sensitivity of individuals to oxidative stress. This method utilizes luminol-amplified chemiluminescence to quantify production of reactive oxygen species (ROS) by opsonized zymosan-stimulated neutrophils that have been subjected to short-term stress via heat shock. In this study, the chemiluminescence reaction was used to monitor the dynamics of ROS production in neutrophils derived from 17 patients of different ages and genders before and after these neutrophils were subjected to heat shock. In addition, we determined expression of Toll-like receptors using fluorescent-labeled antibody. The effects of adrenaline, dexamethasone, aspirin, and indomethacin, as well as different doses of exogenous heat shock protein 70 (Hsp70), on the production of ROS by stimulated neutrophils was also investigated. Our data showed that adrenaline and exogenous Hsp70 both suppressed ROS production by stimulated neutrophils. Furthermore, TLR4 expression was upregulated upon heat stress. Thus, adrenaline, HSPs, and TLRs may all play a role in regulating stress responses in phagocytes.