Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1972 Jun 1;135(6):1392–1405. doi: 10.1084/jem.135.6.1392

CELL SURFACE IMMUNOGLOBULIN

III. ISOLATION AND CHARACTERIZATION OF IMMUNOGLOBULIN FROM NONSECRETORY HUMAN LYMPHOID CELLS

Charles J Sherr 1, Sonia Baur 1, Inge Grundke 1, Joseph Zeligs 1, Barbara Zeligs 1, Jonathan W Uhr 1
PMCID: PMC2139180  PMID: 5025439

Abstract

Cells from an established line of Burkitt lymphoma (Daudi) were enzymatically radioiodinated, and labeled Ig from the cell surface was isolated and studied. Subcellular fractionation of labeled cells confirmed that intracellular proteins from the cytoplasm are not iodinated by this method. Radioactive Ig was identified as monomeric (8S) IgM, and an average of 105 Ig molecules was found per cell. Ig molecules could be released from the plasma membrane by detergent lysis under nonreducing conditions indicating that attachment of Ig to the plasma membrane occurs via noncovalent interactions. The ratio of µ/L radioactivity in surface Ig was the same as that of total cellular Ig radioiodinated in solution suggesting that a large portion of the Fc fragment is not buried within the membrane. In contrast to the results obtained with cell surface Ig, most intracellular Ig was found as "free" µ- and L chains regardless of whether lysates were labeled with 125I or cells were labeled with leucine-3H. The results indicate that only a small percentage of the total Ig of Daudi cells is associated with the cell surface and suggest that covalent assembly of Ig occurs at or near the time that the molecule becomes part of the plasma membrane. Similarities between cell surface Ig on normal splenic lymphocytes and Daudi cells suggest that the latter is a neoplasm of bone marrow-derived lymphocytes.

Full Text

The Full Text of this article is available as a PDF (752.4 KB).

Selected References

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

  1. Baur S., Schenkein I., Uhr J. W. Cell surface immunoglobulin. I. Isolation and characterization of immunoglobulin from murine myeloma cells. J Immunol. 1972 Mar;108(3):748–754. [PubMed] [Google Scholar]
  2. Baur S., Vitetta E. S., Sherr C. J., Schenkein I., Uhr J. W. Isolation of heavy and light chains of immunoglobulin from the surfaces of lymphoid cells. J Immunol. 1971 Apr;106(4):1133–1135. [PubMed] [Google Scholar]
  3. Buxbaum J., Zolla S., Scharff M. D., Franklin E. C. Synthesis and assembly of immunoglobulins by malignant human plasmacytes and lymphocytes. II. Heterogeneity of assembly in cells producing IgM proteins. J Exp Med. 1971 May 1;133(5):1118–1130. doi: 10.1084/jem.133.5.1118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DALTON A. J., FELIX M. D. A comparison of mesothelial cells and macrophages in mice after the intraperitoneal inoculation of melanin granules. J Biophys Biochem Cytol. 1956 Jul 25;2(4 Suppl):109–114. doi: 10.1083/jcb.2.4.109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Dingman C. W., Peacock A. C. Analytical studies on nuclear ribonucleic acid using polyacrylamide gel electrophoresis. Biochemistry. 1968 Feb;7(2):659–668. doi: 10.1021/bi00842a022. [DOI] [PubMed] [Google Scholar]
  6. Eskeland T., Klein E., Inoue M., Johansson B. Characterization of immunoglobulin structures from the surface of chronic lymphocytic leukemia cells. J Exp Med. 1971 Jul 1;134(1):265–280. doi: 10.1084/jem.134.1.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eskeland T., Klein E. Isolation of 7S IgM and kappa chains from the surface membrane of tissue culture cells derived from a Burkitt lymphoma. J Immunol. 1971 Nov;107(5):1368–1375. [PubMed] [Google Scholar]
  8. FLECK A., MUNRO H. N. The precision of ultraviolet absorption measurements in the Schmidt-Thannhauser procedure for nucleic acid estimation. Biochim Biophys Acta. 1962 May 14;55:571–583. doi: 10.1016/0006-3002(62)90836-3. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. 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]
  11. 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]
  12. Klein E., Eskeland T., Inoue M., Strom R., Johansson B. Surface immunoglobulin-moieties on lymphoid cells. Exp Cell Res. 1970 Sep;62(1):133–148. doi: 10.1016/0014-4827(79)90515-9. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. 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]
  16. LUFT J. H. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. doi: 10.1083/jcb.9.2.409. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lerner R. A., McConahey P. J., Jansen I., Dixon F. J. Synthesis of plasma membrane-associated and secretory immunoglobulin in diploid lymphocytes. J Exp Med. 1972 Jan;135(1):136–149. doi: 10.1084/jem.135.1.136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marchalonis J. J. An enzymic method for the trace iodination of immunoglobulins and other proteins. Biochem J. 1969 Jun;113(2):299–305. doi: 10.1042/bj1130299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Phillips D. R., Morrison M. The arrangement of proteins in the human erythrocyte membrane. Biochem Biophys Res Commun. 1970 Jul 27;40(2):284–289. doi: 10.1016/0006-291x(70)91007-7. [DOI] [PubMed] [Google Scholar]
  21. Rabellino E., Colon S., Grey H. M., Unanue E. R. Immunoglobulins on the surface of lymphocytes. I. Distribution and quantitation. J Exp Med. 1971 Jan 1;133(1):156–167. doi: 10.1084/jem.133.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rowden G. A method of obtaining stained electron-microscope specimens free from contamination. J Microsc. 1969;89(2):229–231. doi: 10.1111/j.1365-2818.1969.tb00669.x. [DOI] [PubMed] [Google Scholar]
  23. STEMPAK J. G., WARD R. T. AN IMPROVED STAINING METHOD FOR ELECTRON MICROSCOPY. J Cell Biol. 1964 Sep;22:697–701. doi: 10.1083/jcb.22.3.697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. 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]
  26. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  27. Sherr C. J., Schenkein I., Uhr J. W. Synthesis and intracellular transport of immunoglobulin in secretory and nonsecretory cells. Ann N Y Acad Sci. 1971 Dec 31;190:250–267. doi: 10.1111/j.1749-6632.1971.tb13539.x. [DOI] [PubMed] [Google Scholar]
  28. Sherr C. J., Uhr J. W. Immunoglobulin synthesis and secretion. VI. Synthesis and intracellular transport of immunoglobulin in nonsecretory lymphoma cells. J Exp Med. 1971 Apr 1;133(4):901–920. doi: 10.1084/jem.133.4.901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Suzuki T., Deutsch H. F. Dissociation, reaggregation, and subunit structure studies of some human gamma-M-globulins. J Biol Chem. 1967 Jun 10;242(11):2725–2738. [PubMed] [Google Scholar]
  30. 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]
  31. Touster O., Aronson N. N., Jr, Dulaney J. T., Hendrickson H. Isolation of rat liver plasma membranes. Use of nucleotide pyrophosphatase and phosphodiesterase I as marker enzymes. J Cell Biol. 1970 Dec;47(3):604–618. doi: 10.1083/jcb.47.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. VENABLE J. H., COGGESHALL R. A SIMPLIFIED LEAD CITRATE STAIN FOR USE IN ELECTRON MICROSCOPY. J Cell Biol. 1965 May;25:407–408. doi: 10.1083/jcb.25.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Vassalli P., Lisowska-Bernstein B., Lamm M. E. Cell-free synthesis of rat immunoglobulin. 3. Analysis of the cell-free made chains and of their mode of assembly. J Mol Biol. 1971 Feb 28;56(1):1–19. doi: 10.1016/0022-2836(71)90080-5. [DOI] [PubMed] [Google Scholar]
  34. Vitetta E. S., Baur S., Uhr J. W. Cell surface immunoglobulin. II. Isolation and characterization of immunoglobulin from mouse splenic lymphocytes. J Exp Med. 1971 Jul 1;134(1):242–264. doi: 10.1084/jem.134.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES