Abstract
Pneumocystis carinii cysts were separated as enriched populations from suspensions of lung homogenate obtained from infected rats containing all developmental stages of this organism. Isolation of cyst populations was achieved by incubating filtered lung homogenate with the fluorescent phospholipid analog 1-palmitoyl-2-C6-NBD-phosphatidylcholine. Whereas cysts did not fluorescence as a result of outer-wall restraint of lipid integration, trophozoites and young intermediate stages readily incorporated the fluorescently labeled lipid analog into their outer membrane. The two distinct labeling patterns displayed by cysts and other developmental phases ofP. carinii constitute a novel, easy, and reproducible means of isolating cysts from infected lung homogenate by flow cytometry.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Babcock GF, Alexander JW, Warden GD (1990) Flow cytometric analysis of neutrophil subsets in thermally injured patients developing infections. Clin Immunol Immunopathol 54:117–125
Campbell WG (1972) Ultrastructure ofPneumocystis in human lungs: life cycle of human pneumocystis. Arch Pathol 93:312–324
Cushion MT, Ebbets D (1990) Growth and metabolism ofPneumocystis carinii in axenic culture. J Clin Microbiol 28:1385–1394
Cushion MT, Walzer PD (1984) Growth and serial passage ofPneumocystis carinii in the A549 cell line. Infect Immunol 44:245–251
Cushion MT, Ruffolo JJ, Walzer PD (1988) Analysis of the developmental stages ofPneumocystis carinii in vitro. Lab Invest 58:324–331
De Stefano JA, Cushion MT, Trinkle L, Walzer PD (1989) Lectins as probes toPneumocystis carinii surface glycomplexes. J Protozool 36:65S-66S
De Stefano JA, Cushion MT, Sleight RG, Walzer PD (1990) Analysis ofPneumocystis carinii cyst wall: I. Evidence for an outer surface membrane. J Protozool 37:428–435
Haselton PS, Curry A, Rankin EH (1981)Pneumocystis carinii pneumonia: a light microscopical and ultrastructural study. J Clin Pathol 34:1138–1146
Mills J (1986)Pneumocystis carinii andToxoplasma gondii infections in patients with AIDS. Rev Infect Dis 8:1001–1011
Pagano RE, Sleight RG (1985) Defining lipid transport and pathways in animal cells. Science 229:1051–1057
Schmatz DM, Romancheck MA, Pittarelli LA (1990) Treatment ofPneumocystis carinii pneumonia with 1,3-β-glucan synthesis inhibitors. Proc Natl Acad Sci USA 87:5950–5954
Sleight RG, Abanto MN (1989) Differences in intracellular transport of a flourescent phosphatidylcholine analog in established cell lines. J Cell Science 93:363–374
Sleight RG, Pagano RE (1984) Transport of a fluorescent phosphatidylcholine analog from the plasma membrane to the Golgi apparatus. J Cell Biol 99:742–751
Thalhammer T, Gessl A, Braakman I, Graf J (1989) Separation of hepatocytes of different acinar zones by flow cytometry. Clin Immunol Immunopathol 54:117–125
Walzer PD, Kim CK, Cushion MT (1989)Pneumocystis carinii. In: Walzer PD, Genta RM (eds) Parasitic infections in the compromised host. Marcel Dekker, New York, pp 83–178
Author information
Authors and Affiliations
Additional information
This study was supported by the Medical Research Service, Department of Veterans Affairs, and by Public Health Service contract NO1-AI-72646 and grants AI-25987 and RO1-HL-46653 from the National Institutes of Health
Rights and permissions
About this article
Cite this article
De Stefano, J.A., Sleight, R.G., Babcock, G.F. et al. Isolation ofPneumocystis carinii cysts by flow cytometry. Parasitol Res 78, 179–182 (1992). https://doi.org/10.1007/BF00931724
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00931724