Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1971 Mar 31;133(4):901–920. doi: 10.1084/jem.133.4.901

IMMUNOGLOBULIN SYNTHESIS AND SECRETION

VI. SYNTHESIS AND INTRACELLULAR TRANSPORT OF IMMUNOGLOBULIN IN NONSECRETORY LYMPHOMA CELLS

Charles J Sherr 1, Jonathan W Uhr 1
PMCID: PMC2138957  PMID: 5547061

Abstract

Cells from an established line of Burkitt's lymphoma (Daudi) and a mouse myeloma (P3K) were pulse-labeled in vitro with 3H-leucine, and immunoglobulin was immunologically precipitated from cell lysates and secretions. In contrast to P3K cells, Daudi cells synthesize a small amount of Ig which is not secreted. Subcellular fractionation experiments indicated that Ig of Daudi cells is synthesized on membrane-bound polyribosomes and enters the cisternae of the microsomes. Ig in the microsomes could be labeled with either 3H-galactose or 3H-fucose suggesting that transport proceeds to the Golgi complex. Additional evidence indicates that Ig molecules are transported to the plasma membrane but are not cleaved from the cell surface. These results together with other studies of Burkitt lymphoma cells suggest that the Daudi line may represent a clone of neoplastic cells derived from normal lymphocytes which synthesize but do not secrete Ig. Similarities between lymphoma cells and antigen-binding cells are discussed.

Full Text

The Full Text of this article is available as a PDF (1.0 MB).

Selected References

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

  1. ATTARDI G., COHN M., HORIBATA K., LENNOX E. S. ANTIBODY FORMATION BY RABBIT LYMPH NODE CELLS. II. FURTHER OBSERVATIONS ON THE BEHAVIOR OF SINGLE ANTIBODY-PRODUCING CELLS WITH RESPECT TO THEIR SYNTHETIC CAPACITY AND MORPHOLOGY. J Immunol. 1964 Mar;92:346–355. [PubMed] [Google Scholar]
  2. Abel C. A., Spiegelberg H. L., Grey H. M. The carbohydrate contents of fragments and polypeptide chains of human gamma-G-myeloma proteins of different heavy-chain subclasses. Biochemistry. 1968 Apr;7(4):1271–1278. doi: 10.1021/bi00844a004. [DOI] [PubMed] [Google Scholar]
  3. Avrameas S., Leduc E. H. Detection of simultaneous antibody synthesis in plasma cells and specialized lymphocytes in rabbit lymph nodes. J Exp Med. 1970 Jun 1;131(6):1137–1168. doi: 10.1084/jem.131.6.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. BOAS N. F. Method for the determination of hexosamines in tissues. J Biol Chem. 1953 Oct;204(2):553–563. [PubMed] [Google Scholar]
  5. Biozzi G., Binaghi R. A., Stiffel C., Mouton D. Production of different classes of immunoglobulins by individual cells in the guinea-pig. Immunology. 1969 Mar;16(3):349–359. [PMC free article] [PubMed] [Google Scholar]
  6. Blobel G., Potter V. R. Ribosomes in rat liver: an estimate of the percentage of free and membrane-bound ribosomes interacting with messenger RNA in vivo. J Mol Biol. 1967 Sep 28;28(3):539–542. doi: 10.1016/s0022-2836(67)80103-7. [DOI] [PubMed] [Google Scholar]
  7. Clark S. L., Jr The synthesis and storage of protein by isolated lymphoid cells, examined by autoradiography with the electron microscope. Am J Anat. 1966 Nov;119(3):375–403. doi: 10.1002/aja.1001190304. [DOI] [PubMed] [Google Scholar]
  8. Coffino P., Scharff M. D. Rate of somatic mutation in immunoglobulin production by mouse myeloma cells. Proc Natl Acad Sci U S A. 1971 Jan;68(1):219–223. doi: 10.1073/pnas.68.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. EAGLE H. Amino acid metabolism in mammalian cell cultures. Science. 1959 Aug 21;130(3373):432–437. doi: 10.1126/science.130.3373.432. [DOI] [PubMed] [Google Scholar]
  10. Eylar E. H. On the biological role of glycoproteins. J Theor Biol. 1966 Jan;10(1):89–113. doi: 10.1016/0022-5193(66)90179-2. [DOI] [PubMed] [Google Scholar]
  11. Fahey J. L., Finegold I., Rabson A. S., Manaker R. A. Immunoglobulin synthesis in vitro by established human cell lines. Science. 1966 May 27;152(3726):1259–1261. doi: 10.1126/science.152.3726.1259. [DOI] [PubMed] [Google Scholar]
  12. Finegold I., Fahey J. L., Dutcher T. F. Immunofluorescent studies of immunoglobulins in human lymphoid cells in continuous culture. J Immunol. 1968 Aug;101(2):366–373. [PubMed] [Google Scholar]
  13. Finegold I., Fahey J. L., Granger H. Synthesis of immunoglobulins by human cell lines in tissue culture. J Immunol. 1967 Nov;99(5):839–848. [PubMed] [Google Scholar]
  14. Greaves M. F., Möller E., Möller G. Studies on antigen-binding cells. II. Relationship to antigen-sensitive cells. Cell Immunol. 1970 Oct;1(4):386–403. doi: 10.1016/0008-8749(70)90016-x. [DOI] [PubMed] [Google Scholar]
  15. Greaves M. F., Möller E. Studies on antigen-binding cells. I. The origin of reactive cells. Cell Immunol. 1970 Oct;1(4):372–385. doi: 10.1016/0008-8749(70)90015-8. [DOI] [PubMed] [Google Scholar]
  16. Greaves M. F., Torrigiani G., Roitt I. M. Blocking of the lymphocyte receptor site for cell mediated hypersensitivity and transplantation reactions by anti-light chain sera. Nature. 1969 May 31;222(5196):885–886. doi: 10.1038/222885a0. [DOI] [PubMed] [Google Scholar]
  17. HELMREICH E., KERN M., EISEN H. N. Observations on the mechanism of secretion of gamma-globulins by isolated lymph node cells. J Biol Chem. 1962 Jun;237:1925–1931. [PubMed] [Google Scholar]
  18. HELMREICH E., KERN M., EISEN H. N. The secretion of antibody by isolated lymph node cells. J Biol Chem. 1961 Feb;236:464–473. [PubMed] [Google Scholar]
  19. Hammond E. Ultrastructural-characteristics of surface IgM reactive malignant lymphoid cells. Exp Cell Res. 1970 Mar;59(3):359–370. doi: 10.1016/0014-4827(70)90642-7. [DOI] [PubMed] [Google Scholar]
  20. Harris T. N., Hummeler K., Harris S. Electron microscopic observations on antibody-producing lymph node cells. J Exp Med. 1966 Jan 1;123(1):161–172. doi: 10.1084/jem.123.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hinuma Y., Grace J. T., Jr Cloning of immunoglobulin-producing human leukemic and lymphoma cells in long-term cultures. Proc Soc Exp Biol Med. 1967 Jan;124(1):107–111. doi: 10.3181/00379727-124-31677. [DOI] [PubMed] [Google Scholar]
  22. Horibata K., Harris A. W. Mouse myelomas and lymphomas in culture. Exp Cell Res. 1970 Apr;60(1):61–77. doi: 10.1016/0014-4827(70)90489-1. [DOI] [PubMed] [Google Scholar]
  23. Klein E., Klein G., Nadkarni J. S., Nadkarni J. J., Wigzell H., Clifford P. Surface IgM-kappa specificity on a Burkitt lymphoma cell in vivo and in derived culture lines. Cancer Res. 1968 Jul;28(7):1300–1310. [PubMed] [Google Scholar]
  24. Klein E., Klein G., Nadkarni J. S., Nadkarni J. J., Wigzell H., Clifford P. Surgace IgM specificity on cells derived from a Burkitt's lymphoma. Lancet. 1967 Nov 18;2(7525):1068–1070. doi: 10.1016/s0140-6736(67)90340-6. [DOI] [PubMed] [Google Scholar]
  25. Leduc E. H., Avrameas S., Bouteille M. Ultrastructural localization of antibody in differentiating plasma cells. J Exp Med. 1968 Jan 1;127(1):109–118. doi: 10.1084/jem.127.1.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lisowska-Bernstein B., Lamm M. E., Vassalli P. Synthesis of immunoglobulin heavy and light chains by the free ribosomes of a mouse plasma cell tumor. Proc Natl Acad Sci U S A. 1970 Jun;66(2):425–432. doi: 10.1073/pnas.66.2.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mach B., Koblet H., Gros D. Chemical identification of specific immunoglobulins as the product of a cell-free system from plasmocytoma tumors. Proc Natl Acad Sci U S A. 1968 Feb;59(2):445–452. doi: 10.1073/pnas.59.2.445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mason S., Warner N. L. The immunoglobulin nature of the antigen recognition site on cells mediating transplantation immunity and delayed hypersentivity. J Immunol. 1970 Mar;104(3):762–765. [PubMed] [Google Scholar]
  29. McConnell I., Munro A., Gurner B. W., Coombs R. R. Studies on actively allergized cells. I. The cyto-dynamics and morphology of rosete-forming lymph node cells in mice and inhibition of rosette-formation with antibody to mouse immunoglobulins. Int Arch Allergy Appl Immunol. 1969;35(3):209–227. [PubMed] [Google Scholar]
  30. Melchers F. The attachment site of carbohydrate in a mouse immunoglobulin light chain. Biochemistry. 1969 Mar;8(3):938–947. doi: 10.1021/bi00831a026. [DOI] [PubMed] [Google Scholar]
  31. Minowada J., Klein G., Clifford P., Klein E., Moore G. E. Studies of Burkitt lymphoma cells. I. Establishment of a cell line (B35M) and its characteristics. Cancer. 1967 Sep;20(9):1430–1437. doi: 10.1002/1097-0142(196709)20:9<1430::aid-cncr2820200909>3.0.co;2-h. [DOI] [PubMed] [Google Scholar]
  32. Mitchell G. F., Miller J. F. Cell to cell interaction in the immune response. II. The source of hemolysin-forming cells in irradiated mice given bone marrow and thymus or thoracic duct lymphocytes. J Exp Med. 1968 Oct 1;128(4):821–837. doi: 10.1084/jem.128.4.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nadkarni J. S., Nadkarni J. J., Clifford P., Manolov G., Fenyö E. M., Klein E. Characteristics of new cell lines derived from Burkitt lymphomas. Cancer. 1969 Jan;23(1):64–79. doi: 10.1002/1097-0142(196901)23:1<64::aid-cncr2820230107>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  34. Nossal G. J., Cunningham A., Mitchell G. F., Miller J. F. Cell to cell interaction in the immune response. 3. Chromosomal marker analysis of single antibody-forming cells in reconstituted, irradiated, or thymectomized mice. J Exp Med. 1968 Oct 1;128(4):839–853. doi: 10.1084/jem.128.4.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Osunkoya B. O., McFarlane H., Luzzatto L., Udeozo I. O., Mottram F. C., Williams A. I., Ngu V. A. Immunoglobin synthesis by fresh biopsy cells and established cell lines from Burkitt's lymphoma. Immunology. 1968 Jun;14(6):851–860. [PMC free article] [PubMed] [Google Scholar]
  36. Paraskevas F., Lee S. T., Israels L. G. Absence of gamma-globulin receptors on mouse plasmacytoma cells. Nature. 1970 Jul 25;227(5256):395–397. doi: 10.1038/227395a0. [DOI] [PubMed] [Google Scholar]
  37. Partridge S. M. Filter-paper partition chromatography of sugars: 1. General description and application to the qualitative analysis of sugars in apple juice, egg white and foetal blood of sheep. with a note by R. G. Westall. Biochem J. 1948;42(2):238–250. doi: 10.1042/bj0420238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pernis B., Forni L., Amante L. Immunoglobulin spots on the surface of rabbit lymphocytes. J Exp Med. 1970 Nov;132(5):1001–1018. doi: 10.1084/jem.132.5.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rifkind R. A., Osserman E. F., Hsu K. C., Morgan C. THE INTRACELLULAR DISTRIBUTION OF GAMMA GLOBULIN IN A MOUSE PLASMA CELL TUMOR (X5563) AS REVEALED BY FLUORESCENCE AND ELECTRON MICROSCOPY. J Exp Med. 1962 Oct 1;116(4):423–432. doi: 10.1084/jem.116.4.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. SELL S., GELL P. G. STUDIES ON RABBIT LYMPHOCYTES IN VITRO. I. STIMULATION OF BLAST TRANSFORMATION WITH AN ANTIALLOTYPE SERUM. J Exp Med. 1965 Aug 1;122:423–440. doi: 10.1084/jem.122.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schenkein I., Uhr J. W. Glycosyl transferases for mouse IgG. J Immunol. 1970 Jul;105(1):271–273. [PubMed] [Google Scholar]
  42. Schenkein I., Uhr J. W. Immunoglobulin synthesis and secretion. I. Biosynthetic studies of the addition of the carbohydrate moieties. J Cell Biol. 1970 Jul;46(1):42–51. doi: 10.1083/jcb.46.1.42. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Schubert D., Cohn M. Immunoglobulin biosynthesis. 3. Blocks in defective synthesis. J Mol Biol. 1968 Dec;38(3):273–288. doi: 10.1016/0022-2836(68)90386-0. [DOI] [PubMed] [Google Scholar]
  44. Schubert D., Horibata K. Immunoglobulin biosynthesis. II. Four independently isolated myeloma variants. J Mol Biol. 1968 Dec;38(3):263–271. doi: 10.1016/0022-2836(68)90385-9. [DOI] [PubMed] [Google Scholar]
  45. Schubert D. Immunoglobulin assembly in a mouse myeloma. Proc Natl Acad Sci U S A. 1968 Jun;60(2):683–690. doi: 10.1073/pnas.60.2.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Shapiro A. L., Scharff M. D., Maizel J. V., Jr, Uhr J. W. Polyribosomal synthesis and assembly of the H and L chains of gamma globulin. Proc Natl Acad Sci U S A. 1966 Jul;56(1):216–221. doi: 10.1073/pnas.56.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sherr C. J., Uhr J. W. Immunoglobulin synthesis and secretion. 3. Incorporation of glucosamine into immunoglobulin on polyribosomes. Proc Natl Acad Sci U S A. 1969 Sep;64(1):381–387. doi: 10.1073/pnas.64.1.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Sherr C. J., Uhr J. W. Immunoglobulin synthesis and secretion. V. Incorporation of leucine and glucosamine into immunoglobulin on free and bound polyribosomes. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1183–1189. doi: 10.1073/pnas.66.4.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Spragg B. P., Clamp J. R. A study of the carbohydrate present in three type K macroglobulins. Biochem J. 1969 Aug;114(1):57–64. doi: 10.1042/bj1140057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Swenson R. M., Kern M. THE SYNTHESIS AND SECRETION OF gamma-GLOBULINS BY LYMPH NODE CELLS, I. THE MICROSOMAL COMPARTMENTALIZATION OF gamma-GLOBULINS. Proc Natl Acad Sci U S A. 1967 Feb;57(2):417–422. doi: 10.1073/pnas.57.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Takahashi M., Yagi Y., Morre G. E., Pressman D. Pattern of immunoglobulin production in individual cells of human hematopoietic origin in established culture. J Immunol. 1969 May;102(5):1274–1283. [PubMed] [Google Scholar]
  52. Tanigaki N., Yagi Y., Moore G. E., Pressman D. Immunoglobulin production in human leukemia cell lines. J Immunol. 1966 Nov;97(5):634–646. [PubMed] [Google Scholar]
  53. Uhr J. W. Intracellular events underlying synthesis and secretion of immunoglobulin. Cell Immunol. 1970 Jul;1(2):228–244. doi: 10.1016/0008-8749(70)90010-9. [DOI] [PubMed] [Google Scholar]
  54. Uhr J. W., Schenkein I. Immunoglobulin synthesis and secretion, IV. Sites of incorporation of sugars as determined by subcellular fractionation. Proc Natl Acad Sci U S A. 1970 Jul;66(3):952–958. doi: 10.1073/pnas.66.3.952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Vassalli P., Lisowska-Bernstein B., Lamm M. E., Benacerraf B. Studies on cell-free synthesis of rat immunoglobulins, II. Synthesis of immunoglobulin and of antibody to the dinitrophenyl hapten. Proc Natl Acad Sci U S A. 1967 Dec;58(6):2422–2429. doi: 10.1073/pnas.58.6.2422. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Vassalli P. Studies on cell-free synthesis of rat immunoglobulins, I. A cell-free system for protein synthesis prepared from lymph-node microsomal vesicles. Proc Natl Acad Sci U S A. 1967 Nov;58(5):2117–2124. doi: 10.1073/pnas.58.5.2117. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Wakefield J. D., Thorbecke G. J., Old L. J., Boyse E. A. Production of immunoglobulins and their subunits by human tissue culture cell lines. J Immunol. 1967 Aug;99(2):308–319. [PubMed] [Google Scholar]
  58. Warner N. L., Byrt P., Ada G. L. Blocking of the lymphocyte antigen receptor site with anti-immunoglobulin sera in vitro. Nature. 1970 Jun 6;226(5249):942–943. doi: 10.1038/226942a0. [DOI] [PubMed] [Google Scholar]
  59. Zagury D., Uhr J. W., Jamieson J. D., Palade G. E. Immunoglobulin synthesis and secretion. II. Radioautographic studies of sites of addition of carbohydrate moieties and intracellular transport. J Cell Biol. 1970 Jul;46(1):52–63. doi: 10.1083/jcb.46.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES