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. 1994 Sep 13;91(19):9131–9135. doi: 10.1073/pnas.91.19.9131

Gastric carcinoma: monoclonal epithelial malignant cells expressing Epstein-Barr virus latent infection protein.

S Imai 1, S Koizumi 1, M Sugiura 1, M Tokunaga 1, Y Uemura 1, N Yamamoto 1, S Tanaka 1, E Sato 1, T Osato 1
PMCID: PMC44761  PMID: 8090780

Abstract

In 1000 primary gastric carcinomas, 70 (7.0%) contained Epstein-Barr virus (EBV) genomic sequences detected by PCR and Southern blots. The positive tumors comprised 8 of 9 (89%) undifferentiated lymphoepithelioma-like carcinomas, 27 of 476 (5.7%) poorly differentiated adenocarcinomas, and 35 of 515 (6.8%) moderately to well-differentiated adenocarcinomas. In situ EBV-encoded small RNA 1 hybridization and hematoxylin/eosin staining in adjacent sections showed that the EBV was present in every carcinoma cell but was not significantly present in lymphoid stroma and in normal mucosa. Two-color immunofluorescence and hematoxylin/eosin staining in parallel sections revealed that every keratin-positive epithelial malignant cell expressed EBV-determined nuclear antigen 1 (EBNA1) but did not significantly express CD45+ infiltrating leukocytes. A single fused terminal fragment was detected in each of the EBNA1-expressing tumors, thereby suggesting that the EBV-carrying gastric carcinomas represent clonal proliferation of cells infected with EBV. The carcinoma cells had exclusively EBNA1 but not EBNA2, -3A, -3B, and -3C; leader protein; and latent membrane protein 1 because of methylation. The patients with EBV-carrying gastric carcinoma had elevated serum EBV-specific antibodies. The EBV-specific cellular immunity was not significantly reduced; however, the cytotoxic T-cell target antigens were not expressed. These findings strongly suggest a causal relation between a significant proportion of gastric carcinoma and EBV, and the virus-carrying carcinoma cells may evade immune surveillance.

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Selected References

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

  1. Dimery I. W., Lee J. S., Blick M., Pearson G., Spitzer G., Hong W. K. Association of the Epstein-Barr virus with lymphoepithelioma of the thymus. Cancer. 1988 Jun 15;61(12):2475–2480. doi: 10.1002/1097-0142(19880615)61:12<2475::aid-cncr2820611215>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]
  2. Finke J., Rowe M., Kallin B., Ernberg I., Rosén A., Dillner J., Klein G. Monoclonal and polyclonal antibodies against Epstein-Barr virus nuclear antigen 5 (EBNA-5) detect multiple protein species in Burkitt's lymphoma and lymphoblastoid cell lines. J Virol. 1987 Dec;61(12):3870–3878. doi: 10.1128/jvi.61.12.3870-3878.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Harabuchi Y., Yamanaka N., Kataura A., Imai S., Kinoshita T., Mizuno F., Osato T. Epstein-Barr virus in nasal T-cell lymphomas in patients with lethal midline granuloma. Lancet. 1990 Jan 20;335(8682):128–130. doi: 10.1016/0140-6736(90)90002-m. [DOI] [PubMed] [Google Scholar]
  4. Henle W., Henle G. E., Horwitz C. A. Epstein-Barr virus specific diagnostic tests in infectious mononucleosis. Hum Pathol. 1974 Sep;5(5):551–565. doi: 10.1016/s0046-8177(74)80006-7. [DOI] [PubMed] [Google Scholar]
  5. Herbst H., Dallenbach F., Hummel M., Niedobitek G., Pileri S., Müller-Lantzsch N., Stein H. Epstein-Barr virus latent membrane protein expression in Hodgkin and Reed-Sternberg cells. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4766–4770. doi: 10.1073/pnas.88.11.4766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hu L. F., Minarovits J., Cao S. L., Contreras-Salazar B., Rymo L., Falk K., Klein G., Ernberg I. Variable expression of latent membrane protein in nasopharyngeal carcinoma can be related to methylation status of the Epstein-Barr virus BNLF-1 5'-flanking region. J Virol. 1991 Mar;65(3):1558–1567. doi: 10.1128/jvi.65.3.1558-1567.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Imai S., Usui N., Sugiura M., Osato T., Sato T., Tsutsumi H., Tachi N., Nakata S., Yamanaka T., Chiba S. Epstein-Barr virus genomic sequences and specific antibodies in cerebrospinal fluid in children with neurologic complications of acute and reactivated EBV infections. J Med Virol. 1993 Aug;40(4):278–284. doi: 10.1002/jmv.1890400405. [DOI] [PubMed] [Google Scholar]
  8. Inouye S., Hondo R. Microplate hybridization of amplified viral DNA segment. J Clin Microbiol. 1990 Jun;28(6):1469–1472. doi: 10.1128/jcm.28.6.1469-1472.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Jones J. F., Shurin S., Abramowsky C., Tubbs R. R., Sciotto C. G., Wahl R., Sands J., Gottman D., Katz B. Z., Sklar J. T-cell lymphomas containing Epstein-Barr viral DNA in patients with chronic Epstein-Barr virus infections. N Engl J Med. 1988 Mar 24;318(12):733–741. doi: 10.1056/NEJM198803243181203. [DOI] [PubMed] [Google Scholar]
  10. Khanna R., Burrows S. R., Kurilla M. G., Jacob C. A., Misko I. S., Sculley T. B., Kieff E., Moss D. J. Localization of Epstein-Barr virus cytotoxic T cell epitopes using recombinant vaccinia: implications for vaccine development. J Exp Med. 1992 Jul 1;176(1):169–176. doi: 10.1084/jem.176.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kikuta H., Taguchi Y., Tomizawa K., Kojima K., Kawamura N., Ishizaka A., Sakiyama Y., Matsumoto S., Imai S., Kinoshita T. Epstein-Barr virus genome-positive T lymphocytes in a boy with chronic active EBV infection associated with Kawasaki-like disease. Nature. 1988 Jun 2;333(6172):455–457. doi: 10.1038/333455a0. [DOI] [PubMed] [Google Scholar]
  12. Kimura H., Futamura M., Kito H., Ando T., Goto M., Kuzushima K., Shibata M., Morishima T. Detection of viral DNA in neonatal herpes simplex virus infections: frequent and prolonged presence in serum and cerebrospinal fluid. J Infect Dis. 1991 Aug;164(2):289–293. doi: 10.1093/infdis/164.2.289. [DOI] [PubMed] [Google Scholar]
  13. Klein G., Dombos L., Gothoskar B. Sensitivity of Epstein-Barr virus (EBV) producer and non-producer human lymphoblastoid cell lines to superinfection with EB-virus. Int J Cancer. 1972 Jul 15;10(1):44–57. doi: 10.1002/ijc.2910100108. [DOI] [PubMed] [Google Scholar]
  14. Klein G., Lindahl T., Jondal M., Leibold W., Menézes J., Nilsson K., Sundström C. Continuous lymphoid cell lines with characteristics of B cells (bone-marrow-derived), lacking the Epstein-Barr virus genome and derived from three human lymphomas. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3283–3286. doi: 10.1073/pnas.71.8.3283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kondo K., Hayakawa Y., Mori H., Sato S., Kondo T., Takahashi K., Minamishima Y., Takahashi M., Yamanishi K. Detection by polymerase chain reaction amplification of human herpesvirus 6 DNA in peripheral blood of patients with exanthem subitum. J Clin Microbiol. 1990 May;28(5):970–974. doi: 10.1128/jcm.28.5.970-974.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Leoncini L., Vindigni C., Megha T., Funtò I., Pacenti L., Musarò M., Renieri A., Seri M., Anagnostopoulos J., Tosi P. Epstein-Barr virus and gastric cancer: data and unanswered questions. Int J Cancer. 1993 Apr 1;53(6):898–901. doi: 10.1002/ijc.2910530605. [DOI] [PubMed] [Google Scholar]
  17. Leyvraz S., Henle W., Chahinian A. P., Perlmann C., Klein G., Gordon R. E., Rosenblum M., Holland J. F. Association of Epstein-Barr virus with thymic carcinoma. N Engl J Med. 1985 May 16;312(20):1296–1299. doi: 10.1056/NEJM198505163122006. [DOI] [PubMed] [Google Scholar]
  18. Masucci M. G., Contreras-Salazar B., Ragnar E., Falk K., Minarovits J., Ernberg I., Klein G. 5-Azacytidine up regulates the expression of Epstein-Barr virus nuclear antigen 2 (EBNA-2) through EBNA-6 and latent membrane protein in the Burkitt's lymphoma line rael. J Virol. 1989 Jul;63(7):3135–3141. doi: 10.1128/jvi.63.7.3135-3141.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Min K. W., Holmquist S., Peiper S. C., O'Leary T. J. Poorly differentiated adenocarcinoma with lymphoid stroma (lymphoepithelioma-like carcinomas) of the stomach. Report of three cases with Epstein-Barr virus genome demonstrated by the polymerase chain reaction. Am J Clin Pathol. 1991 Aug;96(2):219–227. doi: 10.1093/ajcp/96.2.219. [DOI] [PubMed] [Google Scholar]
  20. Minarovits J., Minarovits-Kormuta S., Ehlin-Henriksson B., Falk K., Klein G., Ernberg I. Host cell phenotype-dependent methylation patterns of Epstein-Barr virus DNA. J Gen Virol. 1991 Jul;72(Pt 7):1591–1599. doi: 10.1099/0022-1317-72-7-1591. [DOI] [PubMed] [Google Scholar]
  21. Moss D. J., Rickinson A. B., Pope J. H. Long-term T-cell-mediated immunity to Epstein-Barr virus in man. I. Complete regression of virus-induced transformation in cultures of seropositive donor leukocytes. Int J Cancer. 1978 Dec;22(6):662–668. doi: 10.1002/ijc.2910220604. [DOI] [PubMed] [Google Scholar]
  22. Murray R. J., Kurilla M. G., Brooks J. M., Thomas W. A., Rowe M., Kieff E., Rickinson A. B. Identification of target antigens for the human cytotoxic T cell response to Epstein-Barr virus (EBV): implications for the immune control of EBV-positive malignancies. J Exp Med. 1992 Jul 1;176(1):157–168. doi: 10.1084/jem.176.1.157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Oda K., Tamaru J., Takenouchi T., Mikata A., Nunomura M., Saitoh N., Sarashina H., Nakajima N. Association of Epstein-Barr virus with gastric carcinoma with lymphoid stroma. Am J Pathol. 1993 Oct;143(4):1063–1071. [PMC free article] [PubMed] [Google Scholar]
  24. Pallesen G., Hamilton-Dutoit S. J., Rowe M., Young L. S. Expression of Epstein-Barr virus latent gene products in tumour cells of Hodgkin's disease. Lancet. 1991 Feb 9;337(8737):320–322. doi: 10.1016/0140-6736(91)90943-j. [DOI] [PubMed] [Google Scholar]
  25. Pittaluga S., Loke S. L., So K. C., Cheung K. N., Ma L. Clonal Epstein-Barr virus in lymphoepithelioma-like carcinoma of the stomach: demonstration of viral genome by in situ hybridization and Southern blot analysis. Mod Pathol. 1992 Nov;5(6):661–664. [PubMed] [Google Scholar]
  26. Raab-Traub N., Flynn K. The structure of the termini of the Epstein-Barr virus as a marker of clonal cellular proliferation. Cell. 1986 Dec 26;47(6):883–889. doi: 10.1016/0092-8674(86)90803-2. [DOI] [PubMed] [Google Scholar]
  27. Reedman B. M., Klein G. Cellular localization of an Epstein-Barr virus (EBV)-associated complement-fixing antigen in producer and non-producer lymphoblastoid cell lines. Int J Cancer. 1973 May;11(3):499–520. doi: 10.1002/ijc.2910110302. [DOI] [PubMed] [Google Scholar]
  28. Rooney C. M., Rickinson A. B., Moss D. J., Lenoir G. M., Epstein M. A. Cell-mediated immunosurveillance mechanisms and the pathogenesis of Burkitt's lymphoma. IARC Sci Publ. 1985;(60):249–264. [PubMed] [Google Scholar]
  29. Rowe D. T., Rowe M., Evan G. I., Wallace L. E., Farrell P. J., Rickinson A. B. Restricted expression of EBV latent genes and T-lymphocyte-detected membrane antigen in Burkitt's lymphoma cells. EMBO J. 1986 Oct;5(10):2599–2607. doi: 10.1002/j.1460-2075.1986.tb04540.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rowe M., Evans H. S., Young L. S., Hennessy K., Kieff E., Rickinson A. B. Monoclonal antibodies to the latent membrane protein of Epstein-Barr virus reveal heterogeneity of the protein and inducible expression in virus-transformed cells. J Gen Virol. 1987 Jun;68(Pt 6):1575–1586. doi: 10.1099/0022-1317-68-6-1575. [DOI] [PubMed] [Google Scholar]
  31. Sample J., Brooks L., Sample C., Young L., Rowe M., Gregory C., Rickinson A., Kieff E. Restricted Epstein-Barr virus protein expression in Burkitt lymphoma is due to a different Epstein-Barr nuclear antigen 1 transcriptional initiation site. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6343–6347. doi: 10.1073/pnas.88.14.6343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Sample J., Henson E. B., Sample C. The Epstein-Barr virus nuclear protein 1 promoter active in type I latency is autoregulated. J Virol. 1992 Aug;66(8):4654–4661. doi: 10.1128/jvi.66.8.4654-4661.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Schaefer B. C., Woisetschlaeger M., Strominger J. L., Speck S. H. Exclusive expression of Epstein-Barr virus nuclear antigen 1 in Burkitt lymphoma arises from a third promoter, distinct from the promoters used in latently infected lymphocytes. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6550–6554. doi: 10.1073/pnas.88.15.6550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Shibata D., Tokunaga M., Uemura Y., Sato E., Tanaka S., Weiss L. M. Association of Epstein-Barr virus with undifferentiated gastric carcinomas with intense lymphoid infiltration. Lymphoepithelioma-like carcinoma. Am J Pathol. 1991 Sep;139(3):469–474. [PMC free article] [PubMed] [Google Scholar]
  35. Shibata D., Weiss L. M. Epstein-Barr virus-associated gastric adenocarcinoma. Am J Pathol. 1992 Apr;140(4):769–774. [PMC free article] [PubMed] [Google Scholar]
  36. Telenti A., Marshall W. F., Smith T. F. Detection of Epstein-Barr virus by polymerase chain reaction. J Clin Microbiol. 1990 Oct;28(10):2187–2190. doi: 10.1128/jcm.28.10.2187-2190.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Watanabe S., Tsugane S., Ohno Y. Prediction of the gastric cancer mortality in 2000 in Japan. Jpn J Cancer Res. 1988 Apr;79(4):439–444. doi: 10.1111/j.1349-7006.1988.tb01611.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weiss L. M., Chen Y. Y., Liu X. F., Shibata D. Epstein-Barr virus and Hodgkin's disease. A correlative in situ hybridization and polymerase chain reaction study. Am J Pathol. 1991 Dec;139(6):1259–1265. [PMC free article] [PubMed] [Google Scholar]
  39. Young L. S., Dawson C. W., Clark D., Rupani H., Busson P., Tursz T., Johnson A., Rickinson A. B. Epstein-Barr virus gene expression in nasopharyngeal carcinoma. J Gen Virol. 1988 May;69(Pt 5):1051–1065. doi: 10.1099/0022-1317-69-5-1051. [DOI] [PubMed] [Google Scholar]
  40. Young L., Alfieri C., Hennessy K., Evans H., O'Hara C., Anderson K. C., Ritz J., Shapiro R. S., Rickinson A., Kieff E. Expression of Epstein-Barr virus transformation-associated genes in tissues of patients with EBV lymphoproliferative disease. N Engl J Med. 1989 Oct 19;321(16):1080–1085. doi: 10.1056/NEJM198910193211604. [DOI] [PubMed] [Google Scholar]

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