Kinetic analysis of the initial ingestion rate of albumin-coated paraffin oil particles by human granulocytes and rabbit alveolar macrophages was undertaken to study the mechanism of action of cations and of heat-labile opsonin on engulfment . The rate of uptake of the particles was stimulated by Ca++ Mg++ Mn++, or Co++ . At high concentrations (> 20 HIM) Ca++ and Mg++ inhibited the rate of ingestion . Treatment of the particles with fresh serum (heat-labile opsonin) also stimulated the rate of ingestion . 126 I -labeled C3 was bound to the particles during opsonization . C3-deficient human serum lacked opsonic activity, which was restored by addition of purified C3 . Normal, C2-deficient, and hereditary angioneurotic edema sera had equivalent opsonic activity . The serum opsonic activity thus involved C3 fixation to the particles by means of the properdin system . Although Mg++ and heat-labile opsonin both accelerated the maximal rates of ingestion of the particles, neither altered the particle concentrations associated with one-half maximal ingestion rates . Opsonization of the particles markedly diminished the concentrations of divalent cations causing both stimulatory and inhibitory effects on ingestion rates and altered the shapes of the cation activation curves . 43 Ca was not bound to the particles during opsonization . The results are consistent with a mechanism whereby divalent cations and heat-labile opsonin activate ingestion by stimulating the work of engulfment rather than by merely enhancing cell-particle affinity, and whereby heat-labile opsonin acts by potentiating the effects of divalent cations.
346Early in this century it was appreciated that fresh serum (1) and divalent cations (2) enhanced the ingestion of particles by mammalian phagocytes . Observations on the morphology of ingestion revealed that the phagocytes spread over the surfaces of particles, maintaining intimate contact with them during engulfment (3) . Explanation for the action of opsonins and of divalent cations on this spreading process was sought in their effects on particle surface wettability and charge, and alterations in particle surfaces that facilitated spreading and deformability by the phagocytes were predicted to accelerate phagocytosis (4-7) . These early theories considered the phagocytes as relatively passive subjects of physical forces . More recent work has shown that phagocytosis requires energy expenditure by the cell (8), and that contractile activity, dependent upon hydrolysis of ATP, may regulate mobility of single cells in general (9) . Therefore, it is possible that particles may elicit spreading by acting on a contractile mechanism, and that opsonins and divalent cations stimulate this process .Ingestion must be studied with intact cells, thus complicating characterization of its basis . Kinetic studies of particle uptake represent one approach to this problem . This report presents kinetic characteristics concerning the interactions of particles, cations, and heat-labile serum opsonin on ingestio...