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. 1977 Jun 1;145(6):1484–1500. doi: 10.1084/jem.145.6.1484

Tissue culture studies in Hodgkin’s disease morphologic, cytogenic, cell surface, and enzymatic properties of cultures derived from splenic tumors

JC Long, PC Zamecnik, AC Aisenberg, L Atkins
PMCID: PMC2180683  PMID: 68093

Abstract

Monolayer and suspension cell cultures prepared from Hodgkin's disease tumors in the spleen were examined microscopically and by cytogenetics, tested for lymphocyte and monocyte cell surface properties, and assayed for enzymes by histochemical and spectrophotometric techniques. Hodgkin's disease monolayer cultures were composed of rapidly proliferating round and polygonal cells that were capable of propagation in vitro for an indefinite period of time. Abnormal aneuploid chromosomes were found in short-term Hodgkin's disease monolayers that had been passaged 16-20 times, and in established cell lines carried in culture longer than 3 yr and passaged more than 200 times. Cells fromHodgkin's disease monolayers contained lysozyme (muramidase), fluoride-resistant alpha naphthol acetate esterase, acid and alkaline phosphatase, and chymotrypsin-like activity. The monolayers did not exhibit specific cell surface markers or phagocytosis. Suspension cultures derived from Hodgkin's disease monolayers were composed of cells with aneuploid karyotypes and similar enzymes. The Hodgkin's disease suspension culture cells had surface receptors for complement and IgGFc, lacked surface or cytoplasmic immunoglobulin, and did not form Erosettes, react with an antithymocyte serum, nor exhibit phagocytosis. Normal monolayer culture cells, derived from adult spleen and human fetal spleen and thymus, were composed of spindle cells with a diploid number of chromosomes that could be carried for only a finite period of time in vitro. Normal cultured cells contained similar esterases and phosphatases, but were devoid of lysozyme and chymotrypsin-like activity. The morphologic, cytogenetic, cell surface, and enzymatic findings indicate that our Hodgkin's disease monolayer and suspension cultures are composed of cells with many properties suggesting an origin from monocytes (macrophages) rather than lymphocytes or fibroblasts. The presence of aneuploid karyotypes is consistent with a neoplastic origin and derivation from a malignant cell of Hodgkin's disease.

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Selected References

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  1. Aisenberg A. C., Bloch K. J. Immunoglobulins on the surface of neoplastic lymphocytes. N Engl J Med. 1972 Aug 10;287(6):272–276. doi: 10.1056/NEJM197208102870603. [DOI] [PubMed] [Google Scholar]
  2. Aisenberg A. C., Bloch K. J., Long J. C., Colvin R. B. Reaction of normal human lymphocytes and chronic lymphocytic leukemia cells with an antithymocyte antiserum. Blood. 1973 Mar;41(3):417–423. [PubMed] [Google Scholar]
  3. Bentwich Z., Douglas S. D., Siegal F. P., Kunkel H. G. Human lymphocyte-sheep erythrocyte rosette formation: some characteristics of the interaction. Clin Immunol Immunopathol. 1973 Jul;1(4):511–522. doi: 10.1016/0090-1229(73)90007-x. [DOI] [PubMed] [Google Scholar]
  4. Boecker W. R., Hossfeld D. K., Gallmeier W. M., Schmidt C. G. Clonal growth of Hodgkin cells. Nature. 1975 Nov 20;258(5532):235–236. doi: 10.1038/258235a0. [DOI] [PubMed] [Google Scholar]
  5. Braylan R. C., Jaffe E. S., Burbach J. W., Frank M. M., Johnson R. E., Berard C. W. Similarities of surface characteristics of neoplastic well-differentiated lymphocytes from solid tissues and from peripheral blood. Cancer Res. 1976 May;36(5):1619–1625. [PubMed] [Google Scholar]
  6. COHN Z. A., BENSON B. THE DIFFERENTIATION OF MONONUCLEAR PHAGOCYTES. MORPHOLOGY, CYTOCHEMISTRY, AND BIOCHEMISTRY. J Exp Med. 1965 Jan 1;121:153–170. doi: 10.1084/jem.121.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. COHN Z. A., WIENER E. THE PARTICULATE HYDROLASES OF MACROPHAGES. I. COMPARATIVE ENZYMOLOGY, ISOLATION, AND PROPERTIES. J Exp Med. 1963 Dec 1;118:991–1008. doi: 10.1084/jem.118.6.991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dorfman R. F., Rice D. F., Mitchell A. D., Kempson R. L., Levine G. Ultrastructural studies of Hodgkin's disease. Natl Cancer Inst Monogr. 1973 May;36:221–238. [PubMed] [Google Scholar]
  9. Dvorak H. F. Metallo-enzymes released from Escherichia coli by osmotic shock. I. Selective depression of enzymes in cells grown in the presence of ethylenediaminetetraacetate. J Biol Chem. 1968 May 25;243(10):2640–2646. [PubMed] [Google Scholar]
  10. Einstein L. P., Schneeberger E. E., Colten H. R. Synthesis of the second component of complement by long-term primary cultures of human monocytes. J Exp Med. 1976 Jan 1;143(1):114–126. doi: 10.1084/jem.143.1.114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Epstein A. L., Herman M. M., Kim H., Dorfman R. F., Kaplan H. S. Biology of the human malignant lymphomas. III. Intracranial heterotransplantation in the nude, athymic mouse. Cancer. 1976 May;37(5):2158–2176. doi: 10.1002/1097-0142(197605)37:5<2158::aid-cncr2820370503>3.0.co;2-f. [DOI] [PubMed] [Google Scholar]
  12. Epstein A. L., Kaplan H. S. Biology of the human malignant lymphomas. I. Establishment in continuous cell culture and heterotransplantation of diffuse histiocytic lymphomas. Cancer. 1974 Dec;34(6):1851–1872. doi: 10.1002/1097-0142(197412)34:6<1851::aid-cncr2820340602>3.0.co;2-4. [DOI] [PubMed] [Google Scholar]
  13. GAREN A., LEVINTHAL C. A fine-structure genetic and chemical study of the enzyme alkaline phosphatase of E. coli. I. Purification and characterization of alkaline phosphatase. Biochim Biophys Acta. 1960 Mar 11;38:470–483. doi: 10.1016/0006-3002(60)91282-8. [DOI] [PubMed] [Google Scholar]
  14. Garvin A. J., Spicer S. S., McKeever P. E. The cytochemical demonstration of intracellular immunoglobulin. In neoplasms of lymphoreticular tissue. Am J Pathol. 1976 Mar;82(3):457–478. [PMC free article] [PubMed] [Google Scholar]
  15. Glick A. D., Leech J. H., Flexner J. M., Collins R. D. Ultrastructural study of Reed-Sternberg cells. Comparison with transformed lymphocytes and histiocytes. Am J Pathol. 1976 Oct;85(1):195–208. [PMC free article] [PubMed] [Google Scholar]
  16. Gordon S., Todd J., Cohn Z. A. In vitro synthesis and secretion of lysozyme by mononuclear phagocytes. J Exp Med. 1974 May 1;139(5):1228–1248. doi: 10.1084/jem.139.5.1228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. HUMMEL B. C. A modified spectrophotometric determination of chymotrypsin, trypsin, and thrombin. Can J Biochem Physiol. 1959 Dec;37:1393–1399. [PubMed] [Google Scholar]
  18. KAPLOW L. S. SIMPLIFIED MYELOPEROXIDASE STAIN USING BENZIDINE DIHYDROCHLORIDE. Blood. 1965 Aug;26:215–219. [PubMed] [Google Scholar]
  19. Kadin M. E., Asbury A. K. Long term cultures of Hodgkin's tissue. A morphologic and radioautographic study. Lab Invest. 1973 Feb;28(2):181–184. [PubMed] [Google Scholar]
  20. LITWACK G. Photometric determination of lysozyme activity. Proc Soc Exp Biol Med. 1955 Jul;89(3):401–403. doi: 10.3181/00379727-89-21824. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Li C. Y., Yam L. T., Crosby W. H. Histochemical characterization of cellular and structural elements of the human spleen. J Histochem Cytochem. 1972 Dec;20(12):1049–1058. doi: 10.1177/20.12.1049. [DOI] [PubMed] [Google Scholar]
  23. Long J. C., Aisenberg A. C., Zamecnik P. C. An antigen in Hodgkin's disease tissue cultures: fluorescent antibody studies. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2285–2289. doi: 10.1073/pnas.71.6.2285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Long J. C., Aisenberg A. C., Zamecnik P. C. An antigen in Hodgkin's disease tissue cultures: radioiodine-labeled antibody studies. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2605–2609. doi: 10.1073/pnas.71.7.2605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Long J. C., Aisenberg A. C., Zamecnik P. C. Chromatographic and electrophoretic analysis of an antigen in Hodgkin's disease tissue cultures. J Natl Cancer Inst. 1977 Feb;58(2):223–227. doi: 10.1093/jnci/58.2.223. [DOI] [PubMed] [Google Scholar]
  26. MOORHEAD P. S., NOWELL P. C., MELLMAN W. J., BATTIPS D. M., HUNGERFORD D. A. Chromosome preparations of leukocytes cultured from human peripheral blood. Exp Cell Res. 1960 Sep;20:613–616. doi: 10.1016/0014-4827(60)90138-5. [DOI] [PubMed] [Google Scholar]
  27. Orenstein N. S., Hammond M. E., Dvorak H. F., Feder J. Esterase and protease activity of purified guinea pig basophil granules. Biochem Biophys Res Commun. 1976 Sep 7;72(1):230–235. doi: 10.1016/0006-291x(76)90984-0. [DOI] [PubMed] [Google Scholar]
  28. Povlsen C. O., Fialkow P. J., Klein E., Klein G., Rygaard J., Wiener F. Growth and antigenic properties of a biopsy-derived Burkitt's lymphoma in thymus-less (nude) mice. Int J Cancer. 1973 Jan 15;11(1):30–39. doi: 10.1002/ijc.2910110105. [DOI] [PubMed] [Google Scholar]
  29. Theofilopoulos A. N., Dixon F. J., Bokisch V. A. Binding of soluble immune complexes to human lymphoblastoid cells. I. Characterization of receptors for IgG Fc and complement and description of the binding mechanism. J Exp Med. 1974 Oct 1;140(4):877–894. doi: 10.1084/jem.140.4.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Theofilopoulos A. N., Wilson C. B., Bokisch V. A., Dixon F. J. Binding of soluble immune complexes to human lymphoblastoid cells. II. Use of Raji cells to detect circulating immune complexes in animal and human sera. J Exp Med. 1974 Nov 1;140(5):1230–1244. doi: 10.1084/jem.140.5.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yam L. T., Li C. Y., Crosby W. H. Cytochemical identification of monocytes and granulocytes. Am J Clin Pathol. 1971 Mar;55(3):283–290. doi: 10.1093/ajcp/55.3.283. [DOI] [PubMed] [Google Scholar]
  32. Yam L. T., Tavassoli M., Jacobs P. Differential characterization of the "reticulum cell" in lymphoreticular neoplasms. Am J Clin Pathol. 1975 Aug;64(2):171–179. doi: 10.1093/ajcp/64.2.171. [DOI] [PubMed] [Google Scholar]
  33. Zamecnik P. C., Long J. C. Growth of cultured cells from patients with Hodgkin's disease and transplantation into nude mice. Proc Natl Acad Sci U S A. 1977 Feb;74(2):754–758. doi: 10.1073/pnas.74.2.754. [DOI] [PMC free article] [PubMed] [Google Scholar]

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