Twelve periapical lesions from symptomatic and asymptomatic patients (six each) were obtained and immediately frozen in liquid nitrogen. Six pulps from unerupted third molars as well as chronically inflamed gingival tissues were also obtained, frozen, and used as negative and positive controls, respectively. The concentration of leukotriene (LT) B4 was determined by reverse phase high pressure liquid chromatography. Representative samples from each group were fixed in formalin, sectioned, and stained with hematoxylin and eosin. Extremely low levels of LTB4 were detected in the uninflamed pulpal samples in comparison to those found in chronically inflamed gingival tissues and periradicular lesions. A significant statistical difference was noted between concentrations of LTB4 in periapical lesions of symptomatic patients and those found in asymptomatic patients and samples of chronically inflamed gingival tissues (p < 0.05). In addition, a positive correlation was found between the presence of symptoms, the concentration of LTB4, and presence of polymorphonuclear leukocytes in symptomatic periapical lesions. The results show presence of high concentrations of LTB4 in symptomatic human periapical lesions.