Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1974 Jul;54(1):83–90. doi: 10.1172/JCI107752

Effects of Phorbol Myristate Acetate on the Metabolism and Ultrastructure of Neutrophils in Chronic Granulomatous Disease

John E Repine 1,2,3,4, James G White 1,2,3,4, C Carlyle Clawson 1,2,3,4, Beulah M Holmes 1,2,3,4
PMCID: PMC301526  PMID: 4366245

Abstract

Previous investigations have demonstrated that phorbol myristate acetate (PMA), the active principle of croton oil, stimulates alterations in normal polymorphonuclear leukocytes (PMN) that resemble closely the changes that develop in the cells after phagocytosis of bacteria. The present study has compared the effects of PMA and heat-killed bacteria on the oxygen uptake, glucose oxidation, nitroblue tetrazolium (NBT) reduction, and ultrastructure of normal neutrophils and PMN from six patients with chronic granulomatous disease (CGD). PMA stimulated oxygen consumption, hexose monophosphate shunt activity, and NBT reduction in normal cells but failed to produce similar effects in CGD neutrophils. However, PMA did induce formation of cytoplasmic vacuoles in the CGD cells similar to those observed in normal neutrophils. The results indicate that PMA is a useful nonparticulate agent for distinguishing between normal and CGD neutrophils and for studying basic mechanisms of phagocytosis in normal and abnormal PMN.

Full text

PDF

Images in this article

86Image
on p.86

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BERENDES H., BRIDGES R. A., GOOD R. A. A fatal granulomatosus of childhood: the clinical study of a new syndrome. Minn Med. 1957 May;40(5):309–312. [PubMed] [Google Scholar]
  2. Baird W. M., Melera P. W., Boutwell R. K. Acrylamide gel electrophoresis studies of the incorporation of cytidine- 3 H into mouse skin RNA at early times after treatment with phorbol esters. Cancer Res. 1972 Apr;32(4):781–788. [PubMed] [Google Scholar]
  3. Belman S., Troll W. The inhibition of croton oil-promoted mouse skin tumorigenesis by steroid hormones. Cancer Res. 1972 Mar;32(3):450–454. [PubMed] [Google Scholar]
  4. CARSON M. J., CHADWICK D. L., BRUBAKER C. A., CLELAND R. S., LANDING B. H. THIRTEEN BOYS WITH PROGRESSIVE SEPTIC GRANULOMATOSIS. Pediatrics. 1965 Mar;35:405–412. [PubMed] [Google Scholar]
  5. COHN Z. A., MORSE S. I. Functional and metabolic properties of polymorphonuclear leucocytes. II. The influence of a lipopolysaccharide endotoxin. J Exp Med. 1960 May 1;111:689–704. doi: 10.1084/jem.111.5.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clawson C. C., White J. G. Platelet interaction with bacteria. I. Reaction phases and effects of inhibitors. Am J Pathol. 1971 Nov;65(2):367–380. [PMC free article] [PubMed] [Google Scholar]
  7. Cooper M. R., McCall C. E., Dechatelet L. R. Stimulation of leukocyte hexose monophosphate shunt activity by ascorbic Acid. Infect Immun. 1971 Jun;3(6):851–853. doi: 10.1128/iai.3.6.851-853.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DeChatelet L. R., Cooper M. R., McCall C. E. Stimulation of the hexose monophosphate shunt in human neutrophils by ascorbic acid: mechanism of action. Antimicrob Agents Chemother. 1972 Jan;1(1):12–16. doi: 10.1128/aac.1.1.12. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. EVANS W. H., KARNOVSKY M. L. The biochemical basis of phagocytosis. IV. Some aspects of carbohydrate metabolism during phagocytosis. Biochemistry. 1962 Jan;1:159–166. doi: 10.1021/bi00907a024. [DOI] [PubMed] [Google Scholar]
  10. Estensen R. D., White J. G., Holmes B. Specific degranulation of human polymorphonuclear leukocytes. Nature. 1974 Mar 22;248(446):347–348. doi: 10.1038/248347a0. [DOI] [PubMed] [Google Scholar]
  11. Estensen R. D., White J. G. Ultrastructural features on the platelet response to phorbol myristate acetate. Am J Pathol. 1974 Mar;74(3):441–452. [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Hecker E. Cocarcinogenic principles from the seed oil of Croton tiglium and from other Euphorbiaceae. Cancer Res. 1968 Nov;28(11):2338–2349. [PubMed] [Google Scholar]
  14. Hecker E. Phorbol esters from croton oil. Chemical nature and biological activities. Naturwissenschaften. 1967 Jun;54(11):282–284. doi: 10.1007/BF00620887. [DOI] [PubMed] [Google Scholar]
  15. Hennings H., Boutwell R. K. Studies on the mechanism of skin tumor promotion. Cancer Res. 1970 Feb;30(2):312–320. [PubMed] [Google Scholar]
  16. Holmes B., Good R. A. Laboratory models of chronic granulomatous disease. J Reticuloendothel Soc. 1972 Aug;12(2):216–237. [PubMed] [Google Scholar]
  17. Holmes B., Page A. R., Good R. A. Studies of the metabolic activity of leukocytes from patients with a genetic abnormality of phagocytic function. J Clin Invest. 1967 Sep;46(9):1422–1432. doi: 10.1172/JCI105634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Holmes B., Park B. H., Malawista S. E., Quie P. G., Nelson D. L., Good R. A. Chronic granulomatous disease in females. N Engl J Med. 1970 Jul 30;283(5):217–221. doi: 10.1056/NEJM197007302830501. [DOI] [PubMed] [Google Scholar]
  19. Holmes B., Quie P. G., Windhorst D. B., Good R. A. Fatal granulomatous disease of childhood. An inborn abnormality of phagocytic function. Lancet. 1966 Jun 4;1(7449):1225–1228. doi: 10.1016/s0140-6736(66)90238-8. [DOI] [PubMed] [Google Scholar]
  20. Karnovsky M. L. Chronic granulomatous disease--pieces of a cellular and molecular puzzle. Fed Proc. 1973 Apr;32(4):1527–1533. [PubMed] [Google Scholar]
  21. LANDING B. H., SHIRKEY H. S. A syndrome of recurrent infection and infiltration of viscera by pigmented lipid histiocytes. Pediatrics. 1957 Sep;20(3):431–438. [PubMed] [Google Scholar]
  22. Lehrer R. I. Effects of colchicine and chloramphenicol on the oxidative metabolism and phagocytic activity of human neutrophils. J Infect Dis. 1973 Jan;127(1):40–48. doi: 10.1093/infdis/127.1.40. [DOI] [PubMed] [Google Scholar]
  23. Park B. H., Holmes B. M., Rodey G. E., Good R. A. Nitroblue-tetrazolium test in children with fatal granulomatous disease and newborn infants. Lancet. 1969 Jan 18;1(7586):157–157. doi: 10.1016/s0140-6736(69)91172-6. [DOI] [PubMed] [Google Scholar]
  24. Puszkin E. G., Zucker M. B. Enhancement of platelet and muscle actomyosin ATPase activity and superprecipitation by the tumour-promotor 12-0-tetradecanoyl-phorbol-13-acetate (PMA). Nat New Biol. 1973 Oct 31;245(148):277–280. doi: 10.1038/newbio245277a0. [DOI] [PubMed] [Google Scholar]
  25. Qualliotine D., DeChatelet L. R., McCall C. E., Cooper M. R. Stimulation of oxidative metabolism in polymorphonuclear leukocytes by catecholamines. J Reticuloendothel Soc. 1972 Mar;11(3):263–276. [PubMed] [Google Scholar]
  26. Quie P. G., White J. G., Holmes B., Good R. A. In vitro bactericidal capacity of human polymorphonuclear leukocytes: diminished activity in chronic granulomatous disease of childhood. J Clin Invest. 1967 Apr;46(4):668–679. doi: 10.1172/JCI105568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Raick A. N., Thumm K., Chivers B. R. Early effects of 12-O-tetradecanoyl-phorbol-13-acetate on the incorporation of tritiated precursor into DNA and the thickness of the interfollicular epidermis, and their relation to tumor promotion in mouse skin. Cancer Res. 1972 Jul;32(7):1562–1568. [PubMed] [Google Scholar]
  28. Romeo D., Zabucchi G., Rossi F. Reversible metabolic stimulation of polymorphonuclear leukocytes and macrophages by concanavalin A. Nat New Biol. 1973 May 23;243(125):111–112. [PubMed] [Google Scholar]
  29. Sivak A., Mossman B. T., Van Duuren B. L. Activation of cell membrane enzymes in the stimulation of cell division. Biochem Biophys Res Commun. 1972 Jan 31;46(2):605–609. doi: 10.1016/s0006-291x(72)80182-7. [DOI] [PubMed] [Google Scholar]
  30. Sivak A., Ray F., Van Duuren B. L. Phorbol ester tumor-promoting agents and membrane stability. Cancer Res. 1969 Mar;29(3):624–630. [PubMed] [Google Scholar]
  31. Sivak A., Van Duuren B. L. Cellular interactions of phorbol myristate acetate in tumor promotion. Chem Biol Interact. 1971 Nov;3(6):401–411. doi: 10.1016/0009-2797(71)90001-9. [DOI] [PubMed] [Google Scholar]
  32. Sivak A., Van Duuren B. L. RNA synthesis induction in cell culture by a tumor promoter. Cancer Res. 1970 Apr;30(4):1203–1205. [PubMed] [Google Scholar]
  33. Van Duuren B. L. Tumor-promoting agents in two-stage carcinogenesis. Prog Exp Tumor Res. 1969;11:31–68. doi: 10.1159/000391388. [DOI] [PubMed] [Google Scholar]
  34. Weissmann G., Troll W., Van Duuren B. L., Sessa G. Studies on lysosomes. X. Effects of tumor-promoting agents upon biological and artificial membrane systems. Biochem Pharmacol. 1968 Dec;17(12):2421–2434. doi: 10.1016/0006-2952(68)90132-9. [DOI] [PubMed] [Google Scholar]
  35. White J. G., Estensen R. D. Cytochemical electron microscopic studies of the action of phorbol myristate acetate on platelets. Am J Pathol. 1974 Mar;74(3):453–466. [PMC free article] [PubMed] [Google Scholar]
  36. White J. G., Estensen R. D. Selective labilization of specific granules in polymorphonuclear leukocytes by phorbol myristate acetate. Am J Pathol. 1974 Apr;75(1):45–60. [PMC free article] [PubMed] [Google Scholar]
  37. White J. G. Giant organelles containing tubules in Chediak-Higashi lymphocytes. Am J Pathol. 1972 Nov;69(2):225–238. [PMC free article] [PubMed] [Google Scholar]
  38. White J. G. The Chediak-Higashi syndrome: a possible lysosomal disease. Blood. 1966 Aug;28(2):143–156. [PubMed] [Google Scholar]
  39. White J. G. The Chediak-Higashi syndrome: cytoplasmic sequestration in circulating leukocytes. Blood. 1967 Apr;29(4):435–451. [PubMed] [Google Scholar]
  40. Zucker M. B., Troll W., Belman S. The tumor-promoter phorbol ester (12-O-tetradecanoyl-phorbol-13-acetate), a potent aggregating agent for blood platelets. J Cell Biol. 1974 Feb;60(2):325–336. doi: 10.1083/jcb.60.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES