Journal of Immunological Methods, 98 (1987) 113-118 113 Elsevier JIM 04275 Uptake of 3H-deoxyglucose as a microassay of human neutrophil and monocyte activation W.K. Scow, S.E. Smith, J.G. McCormack and Y.H. Thong Immunobiology Laboratory, Departments of Child Health and Medicine, University of Queensland, Mater Public Hospitals, South Brisbane 4101, Australia (Received24 July1986,revisedreceived10 November1986,accepted 4 December1986) The accumulation of 2-deoxy-D-[1-3H]glucose, a non-metabolised analogue of glucose provides a quantitative measurement of the state of activation of phagocytic cells. A microassay for 3H-deoxyglucose uptake by human neutrophils and monocytes is described. Optimal conditions for the assay include the use of 5 × 105 cells and 0.78 #Ci/ml of deoxyglucose in a final volume of 0.2 ml per microtitre well, and an incubation time of 30 min at 37 ° C. This simple, rapid and reproducible technique may find application in experimental immunology. Key words: Neutrophil; Monocyte; Deoxy[3H]glucose uptake; Diagnostic immunology Introduction Phagocytic cells have important roles in host resistance, inflammation and immunoregulation (Nathan et al., 1980; Weissman et al., 1980; John- son, 1982). The two major types of phagocytes in humans are polymorphonuclear neutrophils and cells of the monocyte-macrophage series. The main energy source for these cells comes from the metabolism of glucose. The glycolytic pathway provides energy for adherence, chemotaxis, phago- cytosis and other vital cell functions (Borregaard and Herlin, 1982), while the hexose monophos- phate shunt provides energy for the generation of oxygen-free radicals which are vital for microbi- cidal activities and mediation of inflammatory tissue destruction (Baehner et al., 1970; Klebanoff, 1975; Babior, 1978). The transport and uptake of glucose into Correspondence to: Y.H. Thong, Immunobiology Labora- tory, Mater Children's Hospital, South Brisbane 4101, Australia. phagocytic cells can be measured by radiometric techniques. For such assays analogues of glucose that are not metabolised should be used so that their accumulation in phagocytic cells provides a quantitative assessment of the state of activation. The 2-deoxy-D-[1-3H]glucose uptake assay has been used for the study of macrophage activation in quinea pigs and other animal species (Bonventre and Mukkada, 1974; Mukkada and Bonventre, 1975; Bonventre et al., 1977; McCormack et al., 1981). We have adapted this method to human neutrophils and monocytes and have described a simple, rapid and reproducible microassay tech- nique suitable for use in experimental immunol- ogy. Materials and methods Reagents 2-Deoxy-o-[1-3H]glucose was purchased from the Radiochemical Centre, Amersham, U.K. It had a specific activity of 15 Ci/mmol or 90.9 0022-1759/87/$03.50 © 1987ElsevierSciencePublishersB.V. (Biomedical Division)