This study was designed to determine the dosimetry within macrophages of beryllium compounds administered at sublethal doses. Information on the dosimetry of beryllium within macrophages is required to guide further efforts to isolate and characterize beryllium-containing haptens. Inhalation of beryllium aerosols can cause chronic berylliosis, a progressive, granulomatous fibrosis of the lung. Studies in laboratory animals indicate that alveolar macrophages take up beryllium compounds and participate in a hypersensitivity immune response to beryllium-containing antigen. Beagle dog macrophage cultures were incubated with 7BeSO4 in solution or with suspensions of 7BeO particles that had been calcined at 500 or 1000 degrees C. Beryllium-7 was measured in fractions collected from cultures after successive centrifugation and filtration steps at 2, 6, 20, and 48 h after addition. An insignificant percentage of BeSO4 was taken up by the cells and did not cause cytotoxicity. Maximum BeO uptake occurred within 6 h, was 60 +/- 6% of added BeO, and was independent of BeO calcination temperature or specific surface area. Approximately 22% of 500 degrees C BeO dissolved within 48 h after addition to cell culture, concurrent with 39% cell killing. Dissolved beryllium remained associated with cells until a cytotoxic concentration was reached (2.2 x 10(-5) M, 15 nmol Be/10(6) cells), when the beryllium was released into the medium. There was no significant dissolution of the 1000 degrees C BeO within 48 h, and no significant cell killing. The results indicate that beryllium dissolved from phagocytized BeO was more cytotoxic than soluble beryllium added extracellularly. The data support an interactive mechanism in which phagocytized BeO particles were dissolved, and dissolved beryllium remained associated with the macrophage until a cytotoxic concentration accumulated, whereupon the beryllium was released to the medium and not appreciably taken up by viable cells.

No abstract

Because of the role that alveolar macrophages (PAM) play in the pulmonary clearance of inhaled particles via mechanical transport and dissolution, understanding the uptake and dissolution of particles by these cells might provide insight into the mechanisms of particle dissolution in lungs of various species and hence facilitate the extrapolation of animal data to humans. Therefore, experiments were conducted to study the phagocytosis and dissolution of 241AmO2 particles by rat, dog and monkey PAM in vitro. Rat, dog and monkey PAM were exposed for up to 72 h to 0.19, 0.93 or 4.6 kBq/ml 241Am, after which cell viability was determined. The 241Am concentration, 4.63 kBq/ml, was used for the phagocytosis and dissolution experiments. The phagocytosis and dissolution of 241AmO2 particles were followed up to 20 and 72 h, respectively. Dog and monkey PAM took up 241AmO2 particles at similar rates, whereas rat PAM phagocytosed only 60% of the amount phagocytosed by dog and monkey PAM at 20 h. The PAM of the three species dissolved 241AmO2 particles at similar rates; 8-10% was dissolved by 72 h. The results of the 241AmO2 uptake in vitro may reflect in vivo situations, where the differences in uptake seen in vitro would probably diminish at later times after exposure. The dissolution results imply that the dissolution of 241AmO2 particles by alveolar macrophages of the three species might be species-independent. This, at least, might be true for dog and monkey, where in vivo data have shown that 241AmO2 was translocated similarly in both species. Finally, the alveolar macrophage culture system provides a useful simulation to investigate uptake and dissolution of inhaled particles.

No abstract

Because alveolar macrophages play a role in the pulmonary clearance of inhaled particles, experiments were conducted to investigate the toxicity of 241AmO2 particles to alveolar macrophages and the role of these macrophages in the dissolution of 241AmO2 particles. Beagle dog pulmonary alveolar macrophages obtained by bronchopulmonary lavage were exposed in vitro to selected concentrations of 241AmO2. Macrophage viability determined by trypan-blue dye exclusion technique and the ability of the alveolar macrophages to phagocytose opsonized sheep red blood cells were the measures of 241AmO2 toxicity. The uptake of 241AmO2 particles was studied as a function of time. In addition, the dissolution of 241AmO2 by macrophages was determined for periods up to 72 h. After 20 h exposure to concentrations of 241Am higher than 4.63 kBq/ml, the phagocytic ability of macrophages was reduced, whereas no significant change in cell viability was observed at this concentration. Significant cell killing occurred at concentrations higher than 18.5 kBq/ml. After 72 h in the cultures, 10% of the 241AmO2 was dissolved by the alveolar macrophages. These findings imply that the inhalation of radioactive particles such as 241AmO2 particles might cause a reduction in the alveolar macrophage population in the lungs. In addition, the dissolution of 241AmO2 particles by alveolar macrophages might play a role in the short-term pulmonary clearance of inhaled 241AmO2 particles in the beagle dog.