Luminol- or Lucigenin-Coated Micropolystyrene Beads, a Single Reagent to Study Opsonin-Independent Phagocytosis by Cellular Chemiluminescence: Reaction with Human Neutrophils, Monocytes, and Differentiated HL60 Cells

"We report that two different micropolystyrene round beads linked to either luminol or lucigenin have been developed. The 1-µ.m-diameter beads arc useful reagents that allow one to perform chemiluminescent (light producing) phagoeytic reactions without opsonization. It is known that luminol reacts with peroxidases (e.g., myeloperoxidase) while lucigenin reacts mainly with superoxide anion. Luminol and lucigenin beads produce cellular chemiluminescence (CL) with neutrophils, macrophages, monocytes, and differentiated human promyelocytic leukemic cells (HL60). When myeloperoxidase (MPO) is present luminol beads evoke higher CL from neutrophils, but when little or no MPO is present, lucigenin beads produce higher amounts of light. Lucigenin beads appear to be active inside these cells where unbound lucigenin does not enter freely.INTRODUCTIONA variety of cells, e.g., neutrophils (Ns) and macrophages (Ms), can engulf foreign particles (1). This engulfing or phagocytic activity is used by the body to kill bacteria and to eliminate foreign particles such as respirable dusts, e.g., coal, silica, or cotton. The neutrophil engulfs or phagocytizes bacteria which are coated with plasma antibodies and/or complement proteins (Fig. 1). The coating or opsonization allows the neutrophil to recognize foreign substances. Usually the absence of opsonization prevents the cell's recognition of a foreign particle and the particle is not phagocytized. Macrophages, particularly the alveolar Oung) cells, do not require opsonization for phagocytosis. Ms should not be selective toward individual dusts because they serve a major role in eliminating a variety of these.Particle size can also be a factor. Usually a particle with a diameter of 0.5-5 µ.m is considered in the respirable range. Larger particles are trapped before they reach the alveoli Oung spaces where oxygen exchange with blood occurs), ·and smaller particles do not generally settle, and are not engulfed by macrophages (2). Tabato and Ikada (3) found that maximal phagocytosis of microspheres takes place when their diameter is in the range of 1.0 to 2.0 µ.m. In addition, they found that beads with hydrophobic surfaces were more readily phagocytized than hydrophilic surfaces."