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. 1992 Nov 1;176(5):1405–1414. doi: 10.1084/jem.176.5.1405

Characterization of an Epstein-Barr virus receptor on human epithelial cells

PMCID: PMC2119421  PMID: 1383386

Abstract

Epstein-Barr virus (EBV) adsorption to human B lymphocytes is mediated by the viral envelope glycoprotein, gp350/220, which binds to the cell surface protein, CD21, also known as the CR2 complement receptor. Human epithelial cells also express an EBV receptor. A candidate surface molecule of 195 kD has previously been identified on an epithelial cell line and explanted epithelial tissue by reactivity with the CD21 specific monoclonal antibody (mAb), HB-5a. In experiments to further characterize the epithelial cell EBV receptor, we have found that two human epithelial cell lines, RHEK-1 and HeLa, specifically bind intact EB virions. A 145-kD protein, similar in size to B lymphocyte CD21, was specifically precipitated from surface iodinated RHEK-1 cells using the HB-5a mAb, or using purified soluble gp350/220 coupled to agarose beads. The previously identified 195-kD protein did not bind to gp350/220 or react with two other anti-CD21 mAbs. CD21 homologous RNA, similar in size to the B lymphocyte CD21 mRNA, was detected in both RHEK-1 and HeLa cells. The nucleotide sequence of the epithelial cell cDNA was identical to B lymphocyte CD21. The longest clone differs from previously reported CD21 cDNAs in having additional 5' untranslated sequence. Polymerase chain reaction amplification of RHEK-1- or B lymphoblastoid-derived cDNA verified that most CD21 transcripts are initiated at least 30-50 nucleotides upstream of the previously reported mRNA cap site. These experiments demonstrate that human epithelial cells can express CD21, and that CD21 is likely to mediate EBV adsorption to epithelial cells.

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Selected References

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  1. Ahearn J. M., Hayward S. D., Hickey J. C., Fearon D. T. Epstein-Barr virus (EBV) infection of murine L cells expressing recombinant human EBV/C3d receptor. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9307–9311. doi: 10.1073/pnas.85.23.9307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Billaud M., Busson P., Huang D., Mueller-Lantzch N., Rousselet G., Pavlish O., Wakasugi H., Seigneurin J. M., Tursz T., Lenoir G. M. Epstein-Barr virus (EBV)-containing nasopharyngeal carcinoma cells express the B-cell activation antigen blast2/CD23 and low levels of the EBV receptor CR2. J Virol. 1989 Oct;63(10):4121–4128. doi: 10.1128/jvi.63.10.4121-4128.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bravo R., Small J. V., Fey S. J., Larsen P. M., Celis J. E. Architecture and polypeptide composition of HeLa cytoskeletons. Modification of cytoarchitectural polypeptides during mitosis. J Mol Biol. 1982 Jan 5;154(1):121–143. doi: 10.1016/0022-2836(82)90421-1. [DOI] [PubMed] [Google Scholar]
  4. Carel J. C., Myones B. L., Frazier B., Holers V. M. Structural requirements for C3d,g/Epstein-Barr virus receptor (CR2/CD21) ligand binding, internalization, and viral infection. J Biol Chem. 1990 Jul 25;265(21):12293–12299. [PubMed] [Google Scholar]
  5. Chen Y. X., Welte K., Gebhard D. H., Evans R. L. Induction of T cell aggregation by antibody to a 16kd human leukocyte surface antigen. J Immunol. 1984 Nov;133(5):2496–2501. [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Cooper N. R., Bradt B. M., Rhim J. S., Nemerow G. R. CR2 complement receptor. J Invest Dermatol. 1990 Jun;94(6 Suppl):112S–117S. doi: 10.1111/1523-1747.ep12876069. [DOI] [PubMed] [Google Scholar]
  8. David E. M., Morgan A. J. Efficient purification of Epstein-Barr virus membrane antigen gp340 by fast protein liquid chromatography. J Immunol Methods. 1988 Apr 6;108(1-2):231–236. doi: 10.1016/0022-1759(88)90424-3. [DOI] [PubMed] [Google Scholar]
  9. Fingeroth J. D., Weis J. J., Tedder T. F., Strominger J. L., Biro P. A., Fearon D. T. Epstein-Barr virus receptor of human B lymphocytes is the C3d receptor CR2. Proc Natl Acad Sci U S A. 1984 Jul;81(14):4510–4514. doi: 10.1073/pnas.81.14.4510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Frohman M. A., Dush M. K., Martin G. R. Rapid production of full-length cDNAs from rare transcripts: amplification using a single gene-specific oligonucleotide primer. Proc Natl Acad Sci U S A. 1988 Dec;85(23):8998–9002. doi: 10.1073/pnas.85.23.8998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fujisaku A., Harley J. B., Frank M. B., Gruner B. A., Frazier B., Holers V. M. Genomic organization and polymorphisms of the human C3d/Epstein-Barr virus receptor. J Biol Chem. 1989 Feb 5;264(4):2118–2125. [PubMed] [Google Scholar]
  12. Greenspan J. S., Greenspan D., Lennette E. T., Abrams D. I., Conant M. A., Petersen V., Freese U. K. Replication of Epstein-Barr virus within the epithelial cells of oral "hairy" leukoplakia, an AIDS-associated lesion. N Engl J Med. 1985 Dec 19;313(25):1564–1571. doi: 10.1056/NEJM198512193132502. [DOI] [PubMed] [Google Scholar]
  13. Jones H. W., Jr, McKusick V. A., Harper P. S., Wuu K. D. George Otto Gey. (1899-1970). The HeLa cell and a reappraisal of its origin. Obstet Gynecol. 1971 Dec;38(6):945–949. [PubMed] [Google Scholar]
  14. Kansas G. S., Tedder T. F. Transmembrane signals generated through MHC class II, CD19, CD20, CD39, and CD40 antigens induce LFA-1-dependent and independent adhesion in human B cells through a tyrosine kinase-dependent pathway. J Immunol. 1991 Dec 15;147(12):4094–4102. [PubMed] [Google Scholar]
  15. Lemon S. M., Hutt L. M., Shaw J. E., Li J. L., Pagano J. S. Replication of EBV in epithelial cells during infectious mononucleosis. Nature. 1977 Jul 21;268(5617):268–270. doi: 10.1038/268268a0. [DOI] [PubMed] [Google Scholar]
  16. Li Q. X., Young L. S., Niedobitek G., Dawson C. W., Birkenbach M., Wang F., Rickinson A. B. Epstein-Barr virus infection and replication in a human epithelial cell system. Nature. 1992 Mar 26;356(6367):347–350. doi: 10.1038/356347a0. [DOI] [PubMed] [Google Scholar]
  17. Lowell C. A., Klickstein L. B., Carter R. H., Mitchell J. A., Fearon D. T., Ahearn J. M. Mapping of the Epstein-Barr virus and C3dg binding sites to a common domain on complement receptor type 2. J Exp Med. 1989 Dec 1;170(6):1931–1946. doi: 10.1084/jem.170.6.1931. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  19. Löning T., Henke R. P., Reichart P., Becker J. In situ hybridization to detect Epstein-Barr virus DNA in oral tissues of HIV-infected patients. Virchows Arch A Pathol Anat Histopathol. 1987;412(2):127–133. doi: 10.1007/BF00716184. [DOI] [PubMed] [Google Scholar]
  20. Martin D. R., Yuryev A., Kalli K. R., Fearon D. T., Ahearn J. M. Determination of the structural basis for selective binding of Epstein-Barr virus to human complement receptor type 2. J Exp Med. 1991 Dec 1;174(6):1299–1311. doi: 10.1084/jem.174.6.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Moore M. D., Cooper N. R., Tack B. F., Nemerow G. R. Molecular cloning of the cDNA encoding the Epstein-Barr virus/C3d receptor (complement receptor type 2) of human B lymphocytes. Proc Natl Acad Sci U S A. 1987 Dec;84(24):9194–9198. doi: 10.1073/pnas.84.24.9194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Moore M. D., DiScipio R. G., Cooper N. R., Nemerow G. R. Hydrodynamic, electron microscopic, and ligand-binding analysis of the Epstein-Barr virus/C3dg receptor (CR2). J Biol Chem. 1989 Dec 5;264(34):20576–20582. [PubMed] [Google Scholar]
  23. 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]
  24. Nemerow G. R., Cooper N. R. Early events in the infection of human B lymphocytes by Epstein-Barr virus: the internalization process. Virology. 1984 Jan 15;132(1):186–198. doi: 10.1016/0042-6822(84)90102-8. [DOI] [PubMed] [Google Scholar]
  25. Nemerow G. R., Houghten R. A., Moore M. D., Cooper N. R. Identification of an epitope in the major envelope protein of Epstein-Barr virus that mediates viral binding to the B lymphocyte EBV receptor (CR2). Cell. 1989 Feb 10;56(3):369–377. doi: 10.1016/0092-8674(89)90240-7. [DOI] [PubMed] [Google Scholar]
  26. Nemerow G. R., Wolfert R., McNaughton M. E., Cooper N. R. Identification and characterization of the Epstein-Barr virus receptor on human B lymphocytes and its relationship to the C3d complement receptor (CR2). J Virol. 1985 Aug;55(2):347–351. doi: 10.1128/jvi.55.2.347-351.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Niedobitek G., Herbst H., Stein H. Epstein-Barr virus/complement receptor and epithelial cells. Lancet. 1989 Jul 8;2(8654):110–110. doi: 10.1016/s0140-6736(89)90357-7. [DOI] [PubMed] [Google Scholar]
  28. Oren R., Takahashi S., Doss C., Levy R., Levy S. TAPA-1, the target of an antiproliferative antibody, defines a new family of transmembrane proteins. Mol Cell Biol. 1990 Aug;10(8):4007–4015. doi: 10.1128/mcb.10.8.4007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Rhim J. S., Jay G., Arnstein P., Price F. M., Sanford K. K., Aaronson S. A. Neoplastic transformation of human epidermal keratinocytes by AD12-SV40 and Kirsten sarcoma viruses. Science. 1985 Mar 8;227(4691):1250–1252. doi: 10.1126/science.2579430. [DOI] [PubMed] [Google Scholar]
  30. Sixbey J. W., Davis D. S., Young L. S., Hutt-Fletcher L., Tedder T. F., Rickinson A. B. Human epithelial cell expression of an Epstein-Barr virus receptor. J Gen Virol. 1987 Mar;68(Pt 3):805–811. doi: 10.1099/0022-1317-68-3-805. [DOI] [PubMed] [Google Scholar]
  31. Takahashi S., Doss C., Levy S., Levy R. TAPA-1, the target of an antiproliferative antibody, is associated on the cell surface with the Leu-13 antigen. J Immunol. 1990 Oct 1;145(7):2207–2213. [PubMed] [Google Scholar]
  32. Tanner J., Weis J., Fearon D., Whang Y., Kieff E. Epstein-Barr virus gp350/220 binding to the B lymphocyte C3d receptor mediates adsorption, capping, and endocytosis. Cell. 1987 Jul 17;50(2):203–213. doi: 10.1016/0092-8674(87)90216-9. [DOI] [PubMed] [Google Scholar]
  33. Tanner J., Whang Y., Sample J., Sears A., Kieff E. Soluble gp350/220 and deletion mutant glycoproteins block Epstein-Barr virus adsorption to lymphocytes. J Virol. 1988 Dec;62(12):4452–4464. doi: 10.1128/jvi.62.12.4452-4464.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tedder T. F., Clement L. T., Cooper M. D. Expression of C3d receptors during human B cell differentiation: immunofluorescence analysis with the HB-5 monoclonal antibody. J Immunol. 1984 Aug;133(2):678–683. [PubMed] [Google Scholar]
  35. Tedder T. F., Klejman G., Schlossman S. F., Saito H. Structure of the gene encoding the human B lymphocyte differentiation antigen CD20 (B1). J Immunol. 1989 Apr 1;142(7):2560–2568. [PubMed] [Google Scholar]
  36. Timens W., Boes A., Vos H., Poppema S. Tissue distribution of the C3d/EBV-receptor: CD21 monoclonal antibodies reactive with a variety of epithelial cells, medullary thymocytes, and peripheral T-cells. Histochemistry. 1991;95(6):605–611. doi: 10.1007/BF00266748. [DOI] [PubMed] [Google Scholar]
  37. Toothaker L. E., Henjes A. J., Weis J. J. Variability of CR2 gene products is due to alternative exon usage and different CR2 alleles. J Immunol. 1989 May 15;142(10):3668–3675. [PubMed] [Google Scholar]
  38. Wang F., Gregory C., Sample C., Rowe M., Liebowitz D., Murray R., Rickinson A., Kieff E. Epstein-Barr virus latent membrane protein (LMP1) and nuclear proteins 2 and 3C are effectors of phenotypic changes in B lymphocytes: EBNA-2 and LMP1 cooperatively induce CD23. J Virol. 1990 May;64(5):2309–2318. doi: 10.1128/jvi.64.5.2309-2318.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weis J. J., Fearon D. T. The identification of N-linked oligosaccharides on the human CR2/Epstein-Barr virus receptor and their function in receptor metabolism, plasma membrane expression, and ligand binding. J Biol Chem. 1985 Nov 5;260(25):13824–13830. [PubMed] [Google Scholar]
  40. Weis J. J., Tedder T. F., Fearon D. T. Identification of a 145,000 Mr membrane protein as the C3d receptor (CR2) of human B lymphocytes. Proc Natl Acad Sci U S A. 1984 Feb;81(3):881–885. doi: 10.1073/pnas.81.3.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Young L. S., Clark D., Sixbey J. W., Rickinson A. B. Epstein-Barr virus receptors on human pharyngeal epithelia. Lancet. 1986 Feb 1;1(8475):240–242. doi: 10.1016/s0140-6736(86)90776-2. [DOI] [PubMed] [Google Scholar]
  42. Young L. S., Dawson C. W., Brown K. W., Rickinson A. B. Identification of a human epithelial cell surface protein sharing an epitope with the C3d/Epstein-Barr virus receptor molecule of B lymphocytes. Int J Cancer. 1989 May 15;43(5):786–794. doi: 10.1002/ijc.2910430508. [DOI] [PubMed] [Google Scholar]
  43. zur Hausen H., Schulte-Holthausen H., Klein G., Henle W., Henle G., Clifford P., Santesson L. EBV DNA in biopsies of Burkitt tumours and anaplastic carcinomas of the nasopharynx. Nature. 1970 Dec 12;228(5276):1056–1058. doi: 10.1038/2281056a0. [DOI] [PubMed] [Google Scholar]

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