Abstract
A method has been developed for measuring the rate of phagocytosis rather than the quantity of particles ingested per cell when the process is virtually complete. The method, which is simpler and more rapid than those described previously, utilizes cellular monolayers, radioactive particles, and short incubation times. Under the conditions described, the rate of uptake of particles by either guinea-pig peritoneal or human blood leukocytes was proportional to both cell concentration and the time of incubation, and was independent of changes in the concentration of particles during the measurement. The particles were retained by the cells for at least 90 min. The most suitable particles so far used have been 32P-labeled Salmonella typhimurium, and acetyl-14C- or methyl-14C-labeled starch particles. The oxidation of 14C-labeled glucose has been studied under the same conditions that were used for the assays of phagocytosis: the greatest increase in formation of 14CO2 from glucose-1-14C occurred a few minutes after the most rapid period of phagocytosis.
Full Text
The Full Text of this article is available as a PDF (676.2 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- BUHLER D. R. A simple scintillation counting technique for assaying C1402 in a Warburg flask. Anal Biochem. 1962 Nov;4:413–417. doi: 10.1016/0003-2697(62)90143-4. [DOI] [PubMed] [Google Scholar]
- Brzuchowska W. Use of radioactive isotopes in studies on phagocytosis in vitro. Nature. 1966 Oct 8;212(5058):210–211. doi: 10.1038/212210b0. [DOI] [PubMed] [Google Scholar]
- COHN Z. A., MORSE S. I. Interactions between rabbit polymorphonuclear leucocytes and staphylococci. J Exp Med. 1959 Sep 1;110:419–443. doi: 10.1084/jem.110.3.419. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carpenter R. R., Barsales P. B. Uptake by mononuclear phagocytes of protein-coated bentonite particles stabilized with a carbodiimide. J Immunol. 1967 Apr;98(4):844–853. [PubMed] [Google Scholar]
- Carpenter R. R. Phagocytosis by guinea pig splenic cells of Escherichia coli and protein-coated bentonite particles labeled with iodine-125. J Immunol. 1966 Jun;96(6):992–999. [PubMed] [Google Scholar]
- Fidalgo B. V., Najjar V. A. The physiological role of the lymphoid system. VI. The stimulatory effect of leucophilic gamma globulin (leucokinin) on the phagocytic activity of human polymorphonuclear leucocyte. Biochemistry. 1967 Nov;6(11):3386–3392. doi: 10.1021/bi00863a008. [DOI] [PubMed] [Google Scholar]
- Graham R. C., Jr, Karnovsky M. J., Shafer A. W., Glass E. A., Karnovsky M. L. Metabolic and morphological observations on the effect of surface-active agents of leukocytes. J Cell Biol. 1967 Mar;32(3):629–647. doi: 10.1083/jcb.32.3.629. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jenkin C. R., Karnovsky M. L., Rowley D. Preparation of an artificial antigen and immunity to mouse typhoid. Immunology. 1967 Oct;13(4):361–372. [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- MARTIN S. P., GREEN R. Methods for the study of surviving leukocytes: A. Preparation of cell suspension. Methods Med Res. 1958;7:136–138. [PubMed] [Google Scholar]
- Malawista S. E., Bodel P. T. The dissociation by colchicine of phagocytosis from increased oxygen consumption in human leukocytes. J Clin Invest. 1967 May;46(5):786–796. doi: 10.1172/JCI105579. [DOI] [PMC free article] [PubMed] [Google Scholar]
- OREN R., FARNHAM A. E., SAITO K., MILOFSKY E., KARNOVSKY M. L. Metabolic patterns in three types of phagocytizing cells. J Cell Biol. 1963 Jun;17:487–501. doi: 10.1083/jcb.17.3.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ROBERTS J., QUASTEL J. H. PARTICLE UPTAKE BY POLYMORPHONUCLEAR LEUCOCYTES AND EHRLICH ASCITES-CARCINOMA CELLS. Biochem J. 1963 Oct;89:150–156. doi: 10.1042/bj0890150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- SBARRA A. J., KARNOVSKY M. L. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J Biol Chem. 1959 Jun;234(6):1355–1362. [PubMed] [Google Scholar]
- Sastry P. S., Hokin L. E. Studies on the role of phospholipids in phagocytosis. J Biol Chem. 1966 Jul 25;241(14):3354–3361. [PubMed] [Google Scholar]
- Weisman R. A., Korn E. D. Phagocytosis of latex beads by Acanthamoeba. I. Biochemical properties. Biochemistry. 1967 Feb;6(2):485–497. doi: 10.1021/bi00854a017. [DOI] [PubMed] [Google Scholar]