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Kinetics of granulocyte phagocytosis: Rate limited by cytoplasmic viscosity and constrained by cell size

机译:Kinetics of granulocyte phagocytosis: Rate limited by cytoplasmic viscosity and constrained by cell size

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AbstractMicromanipulation of yeast particles and blood granulocytes has been used to study the kinetics of single phagocytosis events. The ingestion process was quantitated by observation of sequential adhesion and encapsulation times. Both adherence and encapsulation times were found to increase greatly as the temperature was reduced below 37°C calcium in solution facilitated adhesion of the particle to the phagocyte but not encapsulation; both adhesion and encapsulation processes required a minimum level of plasma components (presumably complement). The general nature of these observations were confirmatiory of previous studies, but this study is unique in that the specific time course of single particle ingestion was quantitated. It was immediately apparent that the phagocytosis process was 100 efficient above the threshold concentrations required for plasma and temperature, but variations in times from cell to cell indicated heterogeneity in the population. The total time for ingestion varied from as low as 2 sec/particle at 37°C to above several min/particle below 15°C. Encapsulation times for particles were normalized by estimates of particle surface areas to establish a specific time/unit area of particle surface: from 0.5 sec/10−8cm2at 37°C to greater than 8 sec/10−8cm2at 15°C. The temperature dependence of the encapsulation time correlated well with the temperature dependence of the “apparent” viscosity for granulocytes measured by micropipet aspiration. As such, the kinetic properties observed in these phagocytosis tests are consistent with a model that both assembly of the contractile system and the displacement of the surface by active contraction in phagocytosis are limited by viscous dissipation in the cell. Based on temperature dependence of the adhesion time, the activation energy associated with “turning on” the contractile system is quite large, with a value of about 31 kcal/mole. Finally, it was found that serial “feeding” of yeast to a single granulocyte satiated the phagocyte only after six to eleven particles had been encapsulated. The number limit to ingestion was directly proportional to inital siz

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