Analytical subcellular fractionation of alveolar macrophages from normal and BCG-vaccinated rabbits with particular reference to heterogeneity of hydrolase-containing granules.
Macrophages were obtained by pulmonary lavage from normal rabbits or rabbits that had developed pulmonary granulomas after receiving intravenous BCG vaccine 2-3 weeks earlier. The cells were disrupted in iso-osmotic sucrose and a low-speed supernatant was fractionated by isopycnic centrifugation on a linear sucrose density gradient. Three populations of hydrolase-containing granules (putative lysosomes) were found in both normal and BCG-induced macrophages. They were distinguished by their different distributions in the gradient and different sensitivities to disruption by digitonin and were termed:type A, containing lysozyme; type B, containing N-acetyl-beta-glucosaminidase, beta-glactosidase, beta-glucuronidase and possibly some lysozyme; type C, containing cathepsin D. Acid phosphatase appeared to be about equally distributed between type B and C granules. Type A and B granules from BCG-induced macrophages showed markedly greater equilibrium density than did those from normal macrophages. Beta-glucuronidase and acid phosphatase had greater specific activity in the induced cells.
D000818:Animals; D001500:BCG Vaccine; D002500:Centrifugation, Isopycnic; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000201:enzymology*; D004072:Digitonin / Q000494:pharmacology; D005260:Female; D006867:Hydrolases / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology*; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000201:enzymology; D011817:Rabbits; D013347:Subcellular Fractions / Q000201:enzymology
D001500:BCG Vaccine; D006867:Hydrolases; D004072:Digitonin