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. 1983 Oct 1;158(4):1021–1033. doi: 10.1084/jem.158.4.1021

Identification of the membrane receptor for the complement fragment C3d by means of a monoclonal antibody

PMCID: PMC2187368  PMID: 6225820

Abstract

The B2 antigen characterized by means of a monoclonal antibody (14) is a 140,000 Mr protein expressed only in certain stages of the differentiation of lymphocytes of the B lineage. Here we examine the relationship between B2 and the membrane complement receptor type 2 (CR2) for the complement fragment C3d (11, 12), which is also associated only with B cells. Both phenotypic markers are distributed in a similar manner among B cell malignancies and, as shown here, among established cell lines. A polypeptide with binding affinity for C3d was isolated from the membrane of B2-positive cells, i.e., tonsil lymphocytes and Raji cells. We found that this C3d-binding protein not only had the same Mr and isoelectric point (pI) as the B2 antigen, but that it was recognized by the monoclonal antibody to B2. However, anti- B2 does not mask the ligand-binding site of CR2 since it does not prevent the interaction of the purified 140,000 Mr polypeptide with immobilized C3d. Rosette formation between tonsil lymphocytes and erythrocyte intermediates bearing C3d was specifically inhibited by anti-B2. In the case of Raji cells, rosette formation was strongly inhibited only when the lymphocytes were sequentially treated with anti- B2 and with a polyclonal antibody against mouse Ig. In short, B2 and CR2 have a similar distribution among normal and malignant cells, have the same Mr and pI under denaturing conditions, and react with a single monoclonal antibody. We conclude that B2 is identical to CR2.

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Selected References

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  1. Barel M., Charriaut C., Frade R. Isolation and characterization of a C3b receptor-like molecule from membranes of a human B lymphoblastoid cell line (Raji). FEBS Lett. 1981 Dec 21;136(1):111–114. doi: 10.1016/0014-5793(81)81225-2. [DOI] [PubMed] [Google Scholar]
  2. Beller D. I., Springer T. A., Schreiber R. D. Anti-Mac-1 selectively inhibits the mouse and human type three complement receptor. J Exp Med. 1982 Oct 1;156(4):1000–1009. doi: 10.1084/jem.156.4.1000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bhan A. K., Nadler L. M., Stashenko P., McCluskey R. T., Schlossman S. F. Stages of B cell differentiation in human lymphoid tissue. J Exp Med. 1981 Sep 1;154(3):737–749. doi: 10.1084/jem.154.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bianco C., Patrick R., Nussenzweig V. A population of lymphocytes bearing a membrane receptor for antigen-antibody-complement complexes. I. Separation and characterization. J Exp Med. 1970 Oct 1;132(4):702–720. doi: 10.1084/jem.132.4.702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chaplin H., Monroe M. C., Lachmann P. J. Further studies of the C3g component of the alpha 2D fragment of human C3. Clin Exp Immunol. 1983 Mar;51(3):639–646. [PMC free article] [PubMed] [Google Scholar]
  6. Cossman J., Jaffe E. S. Distribution of complement receptor subtypes in non-Hodgkin's lymphomas of B-cell origin. Blood. 1981 Jul;58(1):20–26. [PubMed] [Google Scholar]
  7. Crossley L. G., Porter R. R. Purification of the human complement control protein C3b inactivator. Biochem J. 1980 Oct 1;191(1):173–182. doi: 10.1042/bj1910173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dykman T. R., Cole J. L., Iida K., Atkinson J. P. Polymorphism of human erythrocyte C3b/C4b receptor. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1698–1702. doi: 10.1073/pnas.80.6.1698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. EISEN H. N. PREPARATION OF PURIFIED ANTI-2,4-DINITROPHENYL ANTIBODIES. Methods Med Res. 1964;10:94–102. [PubMed] [Google Scholar]
  10. Eden A., Miller G. W., Nussenzweig V. Human lymphocytes bear membrane receptors for C3b and C3d. J Clin Invest. 1973 Dec;52(12):3239–3242. doi: 10.1172/JCI107525. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fearon D. T., Wong W. W. Complement ligand-receptor interactions that mediate biological responses. Annu Rev Immunol. 1983;1:243–271. doi: 10.1146/annurev.iy.01.040183.001331. [DOI] [PubMed] [Google Scholar]
  12. Ferreira A., Eichinger D. A simplified two-dimensional electrophoretic technique. J Immunol Methods. 1981;43(3):291–299. doi: 10.1016/0022-1759(81)90177-0. [DOI] [PubMed] [Google Scholar]
  13. Harrison R. A., Lachmann P. J. The physiological breakdown of the third component of human complement. Mol Immunol. 1980 Jan;17(1):9–20. doi: 10.1016/0161-5890(80)90119-4. [DOI] [PubMed] [Google Scholar]
  14. Iida K., Mornaghi R., Nussenzweig V. Complement receptor (CR1) deficiency in erythrocytes from patients with systemic lupus erythematosus. J Exp Med. 1982 May 1;155(5):1427–1438. doi: 10.1084/jem.155.5.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Inada S., Brown E. J., Gaither T. A., Hammer C. H., Takahashi T., Frank M. M. C3d receptors are expressed on human monocytes after in vitro cultivation. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2351–2355. doi: 10.1073/pnas.80.8.2351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Klaus G. G. Immune complexes and the problem of immunological memory. Ann Immunol (Paris) 1982 Mar-Apr;133C(2):221–227. doi: 10.1016/0769-2625(82)90036-8. [DOI] [PubMed] [Google Scholar]
  17. Lachmann P. J., Pangburn M. K., Oldroyd R. G. Breakdown of C3 after complement activation. Identification of a new fragment C3g, using monoclonal antibodies. J Exp Med. 1982 Jul 1;156(1):205–216. doi: 10.1084/jem.156.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Lambris J. D., Dobson N. J., Ross G. D. Isolation of lymphocyte membrane complement receptor type two (the C3d receptor) and preparation of receptor-specific antibody. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1828–1832. doi: 10.1073/pnas.78.3.1828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Medof M. E., Iida K., Mold C., Nussenzweig V. Unique role of the complement receptor CR1 in the degradation of C3b associated with immune complexes. J Exp Med. 1982 Dec 1;156(6):1739–1754. doi: 10.1084/jem.156.6.1739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. NISONOFF A. ENZYMATIC DIGESTION OF RABBIT GAMMA GLOBULIN AND ANTIBODY AND CHROMATOGRAPHY OF DIGESTION PRODUCTS. Methods Med Res. 1964;10:134–141. [PubMed] [Google Scholar]
  22. Nadler L. M., Stashenko P., Hardy R., van Agthoven A., Terhorst C., Schlossman S. F. Characterization of a human B cell-specific antigen (B2) distinct from B1. J Immunol. 1981 May;126(5):1941–1947. [PubMed] [Google Scholar]
  23. Papamichail M., Gutierrez C., Embling P., Johnson P., Holborow E. J., Pepys M. B. Complement dependence of localisation of aggregated IgG in germinal centres. Scand J Immunol. 1975;4(4):343–347. doi: 10.1111/j.1365-3083.1975.tb02635.x. [DOI] [PubMed] [Google Scholar]
  24. Polley M. J., Müller-Eberhard H. J. Enharncement of the hemolytic activity of the second component of human complement by oxidation. J Exp Med. 1967 Dec 1;126(6):1013–1025. doi: 10.1084/jem.126.6.1013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ross G. D., Lambris J. D., Cain J. A., Newman S. L. Generation of three different fragments of bound C3 with purified factor I or serum. I. Requirements for factor H vs CR1 cofactor activity. J Immunol. 1982 Nov;129(5):2051–2060. [PubMed] [Google Scholar]
  26. Ross G. D., Lambris J. D. Identification of a C3bi-specific membrane complement receptor that is expressed on lymphocytes, monocytes, neutrophils, and erythrocytes. J Exp Med. 1982 Jan 1;155(1):96–110. doi: 10.1084/jem.155.1.96. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ross G. D., Polley M. J., Rabellino E. M., Grey H. M. Two different complement receptors on human lymphocytes. One specific for C3b and one specific for C3b inactivator-cleaved C3b. J Exp Med. 1973 Oct 1;138(4):798–811. doi: 10.1084/jem.138.4.798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Stashenko P., Nadler L. M., Hardy R., Schlossman S. F. Expression of cell surface markers after human B lymphocyte activation. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3848–3852. doi: 10.1073/pnas.78.6.3848. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weiler J. M., Daha M. R., Austen K. F., Fearon D. T. Control of the amplification convertase of complement by the plasma protein beta1H. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3268–3272. doi: 10.1073/pnas.73.9.3268. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. White R. G., Henderson D. C., Eslami M. B., Neilsen K. H. Localization of a protein antigen in the chicken spleen. Effect of various manipulative procedures on the morphogenesis of the germinal centre. Immunology. 1975 Jan;28(1):1–21. [PMC free article] [PubMed] [Google Scholar]

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