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. 1981 Jun 1;153(6):1426–1444. doi: 10.1084/jem.153.6.1426

Identification of leukemia-associated inhibitory activity as acidic isoferritins. A regulatory role for acidic isoferritins in the production of granulocytes and macrophages

PMCID: PMC2186184  PMID: 6972999

Abstract

Acidic isoferritins have been identified as leukemia-associated inhibitory activity (LIA), which suppresses colony and cluster formation of colony-forming unit-granulocyte macrophages from normal donors but not from patients with leukemia. LIA was detected in all ferritin preparations tested, including ferritin isolated from normal heart, spleen, liver, and placental tissues, and from the spleens of patients with chronic myelogenous leukemia and Hodgkin's disease. Purified preparations of LIA were composed almost entirely of acidic isoferritins, as determined by immunoassay, radioimmunoassay, and isoelectric focusing. The inhibitory activity in the LIA and ferritin samples was inactivated by a battery of antisera specific for ferritin, including those prepared against acidic isoferritins from normal heart and spleen tissues from patients with Hodgkin's disease, and those previously absorbed with basic isoferritins. Antisera absorbed with acidic isoferritins did not inactivate the inhibitory activity. Separation of LIA and chronic myelogenous leukemia and normal spleen ferritin by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and isoelectric focusing confirmed that the regions of peak inhibitory activity corresponded in each to an apparent molecular weight of approximately 550,000 and to a pI value of 4.7. Similar physicochemical characteristics included inactivation by methods that dissociate ferritin molecules into subunits and by treatment with trypsin, chymotrypsin, pronase, and periodate. The purified preparations were extremely stable to heat treatment. The glycoprotein nature of the inhibitory activity was substantiated because it bound to concanavalin A-Sepharose and was eluted off by alpha-methyl mannose. Inhibitory activity of the activity of the acidic isoferritins was detected at concentrations as low as 10(-17)-10(-19) M and iron saturation did not appear to be necessary for its action. These results implicate acidic isoferritins in the regulation of normal myelopoiesis and suggest a role for them in the progression of leukemia.

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Selected References

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  1. Aisen P., Listowsky I. Iron transport and storage proteins. Annu Rev Biochem. 1980;49:357–393. doi: 10.1146/annurev.bi.49.070180.002041. [DOI] [PubMed] [Google Scholar]
  2. Arosio P., Adelman T. G., Drysdale J. W. On ferritin heterogeneity. Further evidence for heteropolymers. J Biol Chem. 1978 Jun 25;253(12):4451–4458. [PubMed] [Google Scholar]
  3. Bognacki J., Broxmeyer H. E., Lobue J. Isolation and biochemical characterization of leukemia-associated inhibitory activity that suppresses colony and cluster formation of cells. Biochim Biophys Acta. 1981 Jan 21;672(2):176–190. doi: 10.1016/0304-4165(81)90391-3. [DOI] [PubMed] [Google Scholar]
  4. Bohn H. Isolierung des Placenta-Proteins PP2 und seine Identifizierung als Ferritin. Arch Gynakol. 1973;215(3):263–275. doi: 10.1007/BF00672810. [DOI] [PubMed] [Google Scholar]
  5. Bomford A., Lis Y., McFarlane I. G., Williams R. Variation in the distribution of two human heart ferritin species. Isoferritin profile and subunit composition in normal and iron-overloaded subjects. Biochem J. 1977 Oct 1;167(1):309–312. doi: 10.1042/bj1670309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Broxmeyer H. E., DeSousa M., Smithyman A., Ralph P., Hamilton J., Kurland J. I., Bognacki J. Specificity and modulation of the action of lactoferrin, a negative feedback regulator of myelopoiesis. Blood. 1980 Feb;55(2):324–333. [PubMed] [Google Scholar]
  7. Broxmeyer H. E., Grossbard E., Jacobsen N., Moore M. A. Evidence for a proliferative advantage of human leukemia colony-forming cells in vitro. J Natl Cancer Inst. 1978 Mar;60(3):513–521. doi: 10.1093/jnci/60.3.513. [DOI] [PubMed] [Google Scholar]
  8. Broxmeyer H. E., Grossbard E., Jacobsen N., Moore M. A. Persistence of inhibitory activity against normal bone-marrow cells during remission of acute leukemia. N Engl J Med. 1979 Aug 16;301(7):346–351. doi: 10.1056/NEJM197908163010702. [DOI] [PubMed] [Google Scholar]
  9. Broxmeyer H. E., Jacobsen N., Kurland J., Mendelsohn N., Moore A. S. In vitro suppression of normal granulocytic stem cells by inhibitory activity derived from human leukemia cells. J Natl Cancer Inst. 1978 Mar;60(3):497–511. doi: 10.1093/jnci/60.3.497. [DOI] [PubMed] [Google Scholar]
  10. Broxmeyer H. E., Moore M. A. Communication between white cells and the abnormalities of this in leukemia. Biochim Biophys Acta. 1978 Oct 27;516(2):129–166. doi: 10.1016/0304-419x(78)90006-9. [DOI] [PubMed] [Google Scholar]
  11. Broxmeyer H. E., Pahwa R., Jacobsen N., Pelus L. M., Ralph P., Meyers P. A., Pahwa S., Kapoor N. Specific inhibitory activity against granulocyte-progenitor cells produced by non-T lymphocytes from patients with neutropenia. Exp Hematol. 1980 Mar;8(3):278–297. [PubMed] [Google Scholar]
  12. Broxmeyer H. E., Ralph P., Gilbertson S., Margolis V. B. Induction of leukemia-associated inhibitory activity and bone marrow granulocyte-macrophage progenitor cell alterations during infection with Abelson virus. Cancer Res. 1980 Nov;40(11):3928–3933. [PubMed] [Google Scholar]
  13. Broxmeyer H. E., Ralph P., Margolis V. B., Nakoinz I., Meyers P., Kapoor N., Moore M. A. Characteristics of bone marrow and blood cells in human leukemia that produce leukemia inhibitory activity (LIA). Leuk Res. 1979;3(4):193–203. doi: 10.1016/0145-2126(79)90042-0. [DOI] [PubMed] [Google Scholar]
  14. Broxmeyer H. E., Smithyman A., Eger R. R., Meyers P. A., de Sousa M. Identification of lactoferrin as the granulocyte-derived inhibitor of colony-stimulating activity production. J Exp Med. 1978 Oct 1;148(4):1052–1067. doi: 10.1084/jem.148.4.1052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Catt K., Tregear G. W. Solid-phase radioimmunoassay in antibody-coated tubes. Science. 1967 Dec 22;158(3808):1570–1572. doi: 10.1126/science.158.3808.1570. [DOI] [PubMed] [Google Scholar]
  16. Cragg S. J., Jacobs A., Parry D. H., Wagstaff M., Worwood M. Isoferritins in acute leukaemia. Br J Cancer. 1977 May;35(5):635–642. doi: 10.1038/bjc.1977.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Drysdale J. W., Adelman T. G., Arosio P., Casareale D., Fitzpatrick P., Harzard J. T., Yokota M. Human isoferritins in normal and disease states. Semin Hematol. 1977 Jan;14(1):71–88. [PubMed] [Google Scholar]
  18. GREENWOOD F. C., HUNTER W. M., GLOVER J. S. THE PREPARATION OF I-131-LABELLED HUMAN GROWTH HORMONE OF HIGH SPECIFIC RADIOACTIVITY. Biochem J. 1963 Oct;89:114–123. doi: 10.1042/bj0890114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hancock B. W., Bruce L., May K., Richmond J. Ferritin, a sensitizing substance in the leucocyte migration inhibition test in patients with malignant lymphoma. Br J Haematol. 1979 Oct;43(2):223–233. doi: 10.1111/j.1365-2141.1979.tb03745.x. [DOI] [PubMed] [Google Scholar]
  20. Harrison P. M. Ferritin: an iron-storage molecule. Semin Hematol. 1977 Jan;14(1):55–70. [PubMed] [Google Scholar]
  21. Hazard J. T., Yokota M., Arosio P., Drysdale J. W. Immunologic differences in human isoferritins: implications for immunologic quantitation of serum ferritin. Blood. 1977 Jan;49(1):139–146. [PubMed] [Google Scholar]
  22. Jones B. M., Worwood M. An immunoradiometric assay for the acidic ferritin of human heart: application to human tissues, cells and serum. Clin Chim Acta. 1978 Apr 3;85(1):81–88. doi: 10.1016/0009-8981(78)90104-3. [DOI] [PubMed] [Google Scholar]
  23. Jones B. M., Worwood M., Jacobs A. Serum ferritin in patients with cancer: determination with antibodies to HeLa cell and spleen ferritin. Clin Chim Acta. 1980 Sep 25;106(2):203–214. doi: 10.1016/0009-8981(80)90173-4. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. Lavoie D. J., Marcus D. M., Otsuka S., Listowsky I. Characterization of ferritin from human placenta. Implications for analysis of tissue specificity and microheterogeneity of ferritins. Biochim Biophys Acta. 1979 Aug 28;579(2):359–366. doi: 10.1016/0005-2795(79)90063-1. [DOI] [PubMed] [Google Scholar]
  27. Linder M. C., Munro H. N. Assay of tissue ferritin. Anal Biochem. 1972 Jul;48(1):266–278. doi: 10.1016/0003-2697(72)90189-3. [DOI] [PubMed] [Google Scholar]
  28. Marcelletti J., Furmanski P. A murine model system for the study of the human leukemia-associated inhibitory activity. Blood. 1980 Jul;56(1):134–137. [PubMed] [Google Scholar]
  29. Matzner Y., Hershko C., Polliack A., Konijn A. M., Izak G. Suppressive effect of ferritin on in vitro lymphocyte function. Br J Haematol. 1979 Jul;42(3):345–353. doi: 10.1111/j.1365-2141.1979.tb01142.x. [DOI] [PubMed] [Google Scholar]
  30. Matzner Y., Konijn A. M., Hershko C. Serum ferritin in hematologic malignancies. Am J Hematol. 1980;9(1):13–22. doi: 10.1002/ajh.2830090103. [DOI] [PubMed] [Google Scholar]
  31. Otsuka S., Listowsky I. High-resolution preparative electrochromatography for purification of two subunit types of ferritin. Anal Biochem. 1980 Mar 1;102(2):419–422. doi: 10.1016/0003-2697(80)90176-1. [DOI] [PubMed] [Google Scholar]
  32. Sirivech S., Frieden E., Osaki S. The release of iron from horse spleen ferritin by reduced flavins. Biochem J. 1974 Nov;143(2):311–315. doi: 10.1042/bj1430311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wagstaff M., Worwood M., Jacobs A. Biochemical and immunological characterization of ferritin from leukaemic cells. Br J Haematol. 1980 Jun;45(2):263–274. doi: 10.1111/j.1365-2141.1980.tb07146.x. [DOI] [PubMed] [Google Scholar]
  34. Wagstaff M., Worwood M., Jacobs A. Properties of human tissue isoferritins. Biochem J. 1978 Sep 1;173(3):969–977. doi: 10.1042/bj1730969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Worwood M., Cragg S. J., Wagstaff M., Jacobs A. Binding of human serum ferritin to concanavalin A. Clin Sci (Lond) 1979 Jan;56(1):83–87. doi: 10.1042/cs0560083. [DOI] [PubMed] [Google Scholar]
  36. de Sousa M., Smithyman A., Tan C. Suggested models of ecotaxopathy in lymphoreticular malignancy. A role for iron-binding proteins in the control of lymphoid cell migration. Am J Pathol. 1978 Feb;90(2):497–520. [PMC free article] [PubMed] [Google Scholar]

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