Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1988 Jan 1;167(1):57–72. doi: 10.1084/jem.167.1.57

The low-affinity receptor for IgE (CD23) on B lymphocytes is spatially associated with HLA-DR antigens

PMCID: PMC2188804  PMID: 2961843

Abstract

Two hybridomas that produce the mAbs 135 and 449 B4 were obtained that inhibited the binding of IgE to the Fc epsilon RL/CD23 on the EBV- transformed B cell line RPMI 8866. mAb 135 was obtained from a mouse immunized with RPMI 8866 cells, whereas mAb 449B4 was obtained from a mouse immunized with a partially purified preparation of Fc epsilon RL/CD23 obtained as the eluate of an IgE immunoabsorbent loaded with a soluble extract of RPMI 8866 cells. These two mAbs bound to Fc epsilon RL/CD23- cell lines and precipitated two polypeptides with 36,000 Mr and 28,000 Mr, which were the HLA-DR alpha and beta chains, respectively. Immunoprecipitation with mAb 135 of NP-40 lysates from dithio-bis(succinimidyl propionate) (DSP) crosslinked 125I-labeled RPMI 8866 or normal B cells incubated with rIL-4 showed three polypeptides with 42,000, 36,000, and 28,000 Mr. The 42,000 Mr polypeptide is identical to the Fc epsilon RL/CD23 since it could be precipitated by the anti-Fc epsilon RL/CD23 mAb 25 after resolubilization from the SDS- PAGE gel. Immunoprecipitations of the crosslinked cell extracts carried out with the anti-Fc epsilon RL/CD23 mAb 25 yielded the same three polypeptides. Furthermore, when RPMI 8866 or rIL-4 preincubated normal B cells were solubilized with a digitonin buffer, which prevents the dissociation of noncovalently linked polypeptide complexes, mAb 135 and mAb 25 precipitated complexes composed of three molecules with 42,000, 36,000, and 28,000 Mr. The well-characterized anti-HLA-DR mAb L243 was unable to block the binding of either IgE or mAb 135 to RPMI 8866 cells, although it could immunoprecipitate the complex (HLA-DR-Fc epsilon RL/CD23) from crosslinked cell lysates. Since mAb 135 and L243 were able to both bind the RPMI 8866 cells, it demonstrates that they bind to different epitopes of the HLA-DR complex, the mAb 135 epitope of the HLA-DR molecule being close to the IgE binding site of the Fc epsilon RL/CD23. These data demonstrated that the Fc epsilon RL/CD23 and HLA-DR antigens are spatially associated on the B cell membrane.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Basten A., Miller J. F., Abraham R. Relationship between Fc receptors, antigen-binding sites on T and B cells, and H-2 complex-associated determinants. J Exp Med. 1975 Mar 1;141(3):547–560. doi: 10.1084/jem.141.3.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bonnefoy J. Y., Aubry J. P., Peronne C., Wijdenes J., Banchereau J. Production and characterization of a monoclonal antibody specific for the human lymphocyte low affinity receptor for IgE: CD 23 is a low affinity receptor for IgE. J Immunol. 1987 May 1;138(9):2970–2978. [PubMed] [Google Scholar]
  3. Bonnefoy J. Y., Banchereau J., Aubry J. P., Wijdenes J. A flow cytometric micromethod for the detection of Fc epsilon receptors and IgE binding factors using fluorescent microspheres. J Immunol Methods. 1986 Apr 3;88(1):25–32. doi: 10.1016/0022-1759(86)90047-5. [DOI] [PubMed] [Google Scholar]
  4. Brenner M. B., Trowbridge I. S., Strominger J. L. Cross-linking of human T cell receptor proteins: association between the T cell idiotype beta subunit and the T3 glycoprotein heavy subunit. Cell. 1985 Jan;40(1):183–190. doi: 10.1016/0092-8674(85)90321-6. [DOI] [PubMed] [Google Scholar]
  5. Defrance T., Aubry J. P., Rousset F., Vanbervliet B., Bonnefoy J. Y., Arai N., Takebe Y., Yokota T., Lee F., Arai K. Human recombinant interleukin 4 induces Fc epsilon receptors (CD23) on normal human B lymphocytes. J Exp Med. 1987 Jun 1;165(6):1459–1467. doi: 10.1084/jem.165.6.1459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dickler H. B., Kubicek M. T., Arbeit R. D., Sharrow S. O. Studies on the nature of the relationship between Ia antigens and Fc receptors on murine B lymphocytes. J Immunol. 1977 Jul;119(1):348–354. [PubMed] [Google Scholar]
  7. Dickler H. B., Sachs D. H. Evidence for identity or close association of the Fc receptor of B lymphocytes and alloantigens determined by the Ir region of the H-2 complex. J Exp Med. 1974 Sep 1;140(3):779–796. doi: 10.1084/jem.140.3.779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Due C., Simonsen M., Olsson L. The major histocompatibility complex class I heavy chain as a structural subunit of the human cell membrane insulin receptor: implications for the range of biological functions of histocompatibility antigens. Proc Natl Acad Sci U S A. 1986 Aug;83(16):6007–6011. doi: 10.1073/pnas.83.16.6007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fehlmann M., Peyron J. F., Samson M., Van Obberghen E., Brandenburg D., Brossette N. Molecular association between major histocompatibility complex class I antigens and insulin receptors in mouse liver membranes. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8634–8637. doi: 10.1073/pnas.82.24.8634. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fridman W. H., Rabourdin-Combe C., Neauport-Sautes C., Gisler R. H. Characterization and function of T cell Fc gamma receptor. Immunol Rev. 1981;56:51–88. doi: 10.1111/j.1600-065x.1981.tb01047.x. [DOI] [PubMed] [Google Scholar]
  11. Gonzalez-Molina A., Spiegelberg H. L. Binding of IgE myeloma proteins to human cultured lymphoblastoid cells. J Immunol. 1976 Nov;117(5 PT2):1838–1845. [PubMed] [Google Scholar]
  12. Ishizaka K. Regulation of IgE synthesis. Annu Rev Immunol. 1984;2:159–182. doi: 10.1146/annurev.iy.02.040184.001111. [DOI] [PubMed] [Google Scholar]
  13. Joseph M., Capron A., Ameisen J. C., Capron M., Vorng H., Pancré V., Kusnierz J. P., Auriault C. The receptor for IgE on blood platelets. Eur J Immunol. 1986 Mar;16(3):306–312. doi: 10.1002/eji.1830160318. [DOI] [PubMed] [Google Scholar]
  14. Katz D. H. Regulation of the IgE system: experimental and clinical aspects. Allergy. 1984 Feb;39(2):81–106. doi: 10.1111/j.1398-9995.1984.tb01940.x. [DOI] [PubMed] [Google Scholar]
  15. Kikutani H., Inui S., Sato R., Barsumian E. L., Owaki H., Yamasaki K., Kaisho T., Uchibayashi N., Hardy R. R., Hirano T. Molecular structure of human lymphocyte receptor for immunoglobulin E. Cell. 1986 Dec 5;47(5):657–665. doi: 10.1016/0092-8674(86)90508-8. [DOI] [PubMed] [Google Scholar]
  16. Kisaki T., Huff T. F., Conrad D. H., Yodoi J., Ishizaka K. Monoclonal antibody specific for T cell-derived human IgE binding factors. J Immunol. 1987 May 15;138(10):3345–3351. [PubMed] [Google Scholar]
  17. Krammer P. H., Pernis B. Ia antigens and Fc receptors. Scand J Immunol. 1976;5(3):205–212. doi: 10.1111/j.1365-3083.1976.tb00271.x. [DOI] [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. Martens C. L., Jardieu P., Trounstine M. L., Stuart S. G., Ishizaka K., Moore K. W. Potentiating and suppressive IgE-binding factors are expressed by a single cloned gene. Proc Natl Acad Sci U S A. 1987 Feb;84(3):809–813. doi: 10.1073/pnas.84.3.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Oettgen H. C., Pettey C. L., Maloy W. L., Terhorst C. A T3-like protein complex associated with the antigen receptor on murine T cells. Nature. 1986 Mar 20;320(6059):272–275. doi: 10.1038/320272a0. [DOI] [PubMed] [Google Scholar]
  21. Peterson L. H., Conrad D. H. Fine specificity, structure, and proteolytic susceptibility of the human lymphocyte receptor for IgE. J Immunol. 1985 Oct;135(4):2654–2660. [PubMed] [Google Scholar]
  22. Rabourdin-Combe C., Dorf M. E., Guimezanes A., Fridman W. H. T cell-produced immunoglobulin binding factor (IBF) bears determinants coded for by the I region of the major histocompatibility complex and lacks allogeneic restriction. Eur J Immunol. 1979 Mar;9(3):237–242. doi: 10.1002/eji.1830090313. [DOI] [PubMed] [Google Scholar]
  23. Rijkers G. T., Mosier D. E. Pneumococcal polysaccharides induce antibody formation by human B lymphocytes in vitro. J Immunol. 1985 Jul;135(1):1–4. [PubMed] [Google Scholar]
  24. Samson M., Cousin J. L., Fehlmann M. Cross-linking of insulin receptors to MHC antigens in human B lymphocytes: evidence for selective molecular interactions. J Immunol. 1986 Oct 1;137(7):2293–2298. [PubMed] [Google Scholar]
  25. Schirrmacher V., Halloran P., David C. S. Interactions of Fc receptors with antibodies against Ia antigens and other cell surface components. J Exp Med. 1975 May 1;141(5):1201–1209. doi: 10.1084/jem.141.5.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schreiber A. B., Schlessinger J., Edidin M. Interaction between major histocompatibility complex antigens and epidermal growth factor receptors on human cells. J Cell Biol. 1984 Feb;98(2):725–731. doi: 10.1083/jcb.98.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Spits H., Borst J., Giphart M., Coligan J., Terhorst C., De Vries J. E. HLA-DC antigens can serve as recognition elements for human cytotoxic T lymphocytes. Eur J Immunol. 1984 Apr;14(4):299–304. doi: 10.1002/eji.1830140404. [DOI] [PubMed] [Google Scholar]
  28. Spits H., Borst J., Giphart M., Coligan J., Terhorst C., De Vries J. E. HLA-DC antigens can serve as recognition elements for human cytotoxic T lymphocytes. Eur J Immunol. 1984 Apr;14(4):299–304. doi: 10.1002/eji.1830140404. [DOI] [PubMed] [Google Scholar]
  29. Suemura M., Shiho O., Deguchi H., Yamamura Y., Böttcher I., Kishimoto T. Characterization and isolation of IgE class-specific suppressor factor (IgE-TsF) I. The presence of the binding site(s) for IgE and of the H-2 gene products in IgE-TsF. J Immunol. 1981 Aug;127(2):465–471. [PubMed] [Google Scholar]
  30. Yodoi J., Ishizaka K. Lymphocytes bearing Fc receptors for IgE. I. Presence of human and rat T lymphocytes with Fc epsilon receptors. J Immunol. 1979 Jun;122(6):2577–2583. [PubMed] [Google Scholar]
  31. Yokota T., Otsuka T., Mosmann T., Banchereau J., DeFrance T., Blanchard D., De Vries J. E., Lee F., Arai K. Isolation and characterization of a human interleukin cDNA clone, homologous to mouse B-cell stimulatory factor 1, that expresses B-cell- and T-cell-stimulating activities. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5894–5898. doi: 10.1073/pnas.83.16.5894. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yssel H., Blanchard D., Boylston A., De Vries J. E., Spits H. T cell clones which share T cell receptor epitopes differ in phenotype, function and specificity. Eur J Immunol. 1986 Oct;16(10):1187–1193. doi: 10.1002/eji.1830161002. [DOI] [PubMed] [Google Scholar]
  33. Yssel H., De Vries J. E., Koken M., Van Blitterswijk W., Spits H. Serum-free medium for generation and propagation of functional human cytotoxic and helper T cell clones. J Immunol Methods. 1984 Aug 3;72(1):219–227. doi: 10.1016/0022-1759(84)90450-2. [DOI] [PubMed] [Google Scholar]
  34. Yukawa K., Kikutani H., Owaki H., Yamasaki K., Yokota A., Nakamura H., Barsumian E. L., Hardy R. R., Suemura M., Kishimoto T. A B cell-specific differentiation antigen, CD23, is a receptor for IgE (Fc epsilon R) on lymphocytes. J Immunol. 1987 Apr 15;138(8):2576–2580. [PubMed] [Google Scholar]

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

RESOURCES