Digestive Diseases and Sciences

, Volume 57, Issue 7, pp 1887–1898 | Cite as

Epstein-Barr Virus Infection Is Common in Inflamed Gastrointestinal Mucosa

  • Julie L. Ryan
  • You-Jun Shen
  • Douglas R. Morgan
  • Leigh B. Thorne
  • Shannon C. Kenney
  • Ricardo L. Dominguez
  • Margaret L. Gulley
Original Article
First Online:
Received:
Accepted:

Abstract

Background and Aims

Epstein-Barr virus (EBV) is present in the malignant epithelial cells of 10% of all gastric adenocarcinomas; however, localization of the virus in normal gastrointestinal mucosa is largely unexplored. In the present study, we measured EBV DNA and localized viral gene products in gastritis specimens (n = 89), normal gastric and colonic mucosa (n = 14), Crohn’s disease (n = 9), and ulcerative colitis (n = 11) tissues.

Methods

A battery of sensitive and specific quantitative polymerase chain reactions targeted six disparate regions of the EBV genome: BamH1 W, EBNA1, LMP1, LMP2, BZLF1, and EBER1. EBV infection was localized by EBV-encoded RNA (EBER) in situ hybridization and by immunohistochemical stains for viral latent proteins LMP1 and LMP2 and for viral lytic proteins BMRF1 and BZLF1. B lymphocytes were identified using CD20 immunostains.

Results

EBV DNA was essentially undetectable in normal gastric mucosa but was present in 46% of gastritis lesions, 44% of normal colonic mucosa, 55% of Crohn’s disease, and 64% of ulcerative colitis samples. Levels of EBV DNA exceeded what would be expected based on the numbers of B lymphocytes in inflamed tissues, suggesting that EBV is preferentially localized to inflammatory gastrointestinal lesions. Histochemical staining revealed EBER expression in lymphoid cells of some PCR-positive lesions. The viral lytic viral proteins, BMRF1 and BZLF1, were expressed in lymphoid cells of two ulcerative colitis tissues, both of which had relatively high viral loads by quantitative PCR.

Conclusion

EBV-infected lymphocytes are frequently present in inflamed gastric and colonic mucosa. Active viral replication in some lesions raises the possibility of virus-related perpetuation of gastrointestinal inflammation.

Keywords

Epstein-Barr virus Gastritis Colitis Inflammation Gene expression Viral load 
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Notes

Acknowledgments

The authors thank Sandra Elmore for technical assistance. This study was funded by the University of North Carolina Department of Pathology and Laboratory Medicine with support from the National Institutes of Health: Environmental Pathology Training Grant (T32-ES07017) supporting graduate studies of Julie L. Ryan, PhD, MPH, Cancer Epidemiology Award (K07CA125588) and funds for the UNC Center for Gastrointestinal Biology and Disease (P30 DK 034987) supporting Douglas R. Morgan, MD MPH, and the Alliance for Clinical Trials in Oncology (U10 CA031946), Clinical Translational Science Award (U54 RR024383), and Innovative Technologies for Molecular Analysis of Cancer (R21 CA155543) award supporting Margaret L. Gulley, MD.

Conflict of interest

MLG is a consultant for McKesson, Abbott Laboratories, and Roche Molecular Systems, and serves on the clinical advisory board of Generation Health.

References

  1. 1.
    Correa P, Piazuelo MB, Camargo MC. The future of gastric cancer prevention. Gastric Cancer. 2004;7:9–16.PubMedCrossRefGoogle Scholar
  2. 2.
    Gulley ML, Tang W. Laboratory assays for Epstein-Barr virus-related disease. J Mol Diagn. 2008;10:279–292.PubMedCrossRefGoogle Scholar
  3. 3.
    Imai S, Koizumi S, Sugiura M, et al. Gastric carcinoma: Monoclonal epithelial malignant cells expressing Epstein-Barr virus latent infection protein. Proc Natl Acad Sci USA. 1994;91:9131–9135.PubMedCrossRefGoogle Scholar
  4. 4.
    Gulley ML, Pulitzer DR, Eagan PA, Schneider BG. Epstein-Barr virus infection is an early event in gastric carcinogenesis and is independent of bcl-2 expression and p53 accumulation. Hum Pathol. 1996;27:20–27.PubMedCrossRefGoogle Scholar
  5. 5.
    Zhang Y, Molot R. Severe gastritis secondary to Epstein-Barr viral infection. Unusual presentation of infectious mononucleosis and associated diffuse lymphoid hyperplasia in gastric mucosa. Arch Pathol Lab Med. 2003;127:478–480.PubMedGoogle Scholar
  6. 6.
    Owens SR, Walls A, Krasinskas AM, Rund CR. Epstein-barr virus gastritis: rare or rarely sampled? A case report. Int J Surg Pathol. 2011;19:196–198.PubMedCrossRefGoogle Scholar
  7. 7.
    Toll AD, Malik S, Tuluc M. Ulcerative gastritis secondary to Epstein-Barr viral infection. Dig Dis Sci. 2010;55:218–219.PubMedCrossRefGoogle Scholar
  8. 8.
    Kitayama Y, Honda S, Sugimura H. Epstein-Barr virus-related gastric pseudolymphoma in infectious mononucleosis. Gastrointest Endosc. 2000;52:290–291.PubMedCrossRefGoogle Scholar
  9. 9.
    Babcock GJ, Decker LL, Freeman RB, Thorley-Lawson DA. Epstein-barr virus-infected resting memory B cells, not proliferating lymphoblasts, accumulate in the peripheral blood of immunosuppressed patients. J Exp Med. 1999;190:567–576.PubMedCrossRefGoogle Scholar
  10. 10.
    Dojcinov SD, Venkataraman G, Pittaluga S, et al. Age-related EBV-associated lymphoproliferative disorders in the Western population: a spectrum of reactive lymphoid hyperplasia and lymphoma. Blood. 2011;117:4726–4735.PubMedCrossRefGoogle Scholar
  11. 11.
    Dojcinov SD, Venkataraman G, Raffeld M, Pittaluga S, Jaffe ES. EBV positive mucocutaneous ulcer—a study of 26 cases associated with various sources of immunosuppression. Am J Surg Pathol. 2010;34:405–417.PubMedCrossRefGoogle Scholar
  12. 12.
    Schneider BG, Gulley ML, Eagan P, Bravo JC, Mera R, Geradts J. Loss of p16/CDKN2A tumor suppressor protein in gastric adenocarcinoma is associated with Epstein-Barr virus and anatomic location in the body of the stomach. Hum Pathol. 2000;31:45–50.PubMedCrossRefGoogle Scholar
  13. 13.
    Arikawa J, Tokunaga M, Tashiro Y, et al. Epstein-Barr virus-positive multiple early gastric cancers and dysplastic lesions: a case report. Pathol Int. 1997;47:730–734.PubMedCrossRefGoogle Scholar
  14. 14.
    Yanai H, Takada K, Shimizu N, Mizugaki Y, Tada M, Okita K. Epstein-Barr virus infection in non-carcinomatous gastric epithelium. J Pathol. 1997;183:293–298.PubMedCrossRefGoogle Scholar
  15. 15.
    Arikawa J, Tokunaga M, Satoh E, Tanaka S, Land CE. Morphological characteristics of Epstein-Barr virus-related early gastric carcinoma: a case-control study. Pathol Int. 1997;47:360–367.PubMedCrossRefGoogle Scholar
  16. 16.
    Hungermann D, Muller S, Spieker T, Lisner R, Niedobitek G, Herbst H. Low prevalence of latently Epstein-Barr virus-infected cells in chronic gastritis. Microsc Res Tech. 2001;53:409–413.PubMedCrossRefGoogle Scholar
  17. 17.
    Hirano A, Yanai H, Shimizu N, et al. Evaluation of epstein-barr virus DNA load in gastric mucosa with chronic atrophic gastritis using a real-time quantitative PCR assay. Int J Gastrointest Cancer. 2003;34:87–94.PubMedCrossRefGoogle Scholar
  18. 18.
    Fukayama M, Chong JM, Uozaki H. Pathology and molecular pathology of Epstein-Barr virus-associated gastric carcinoma. Curr Top Microbiol Immunol. 2001;258:91–102.PubMedCrossRefGoogle Scholar
  19. 19.
    Jing X, Nakamura Y, Nakamura M, et al. Detection of Epstein-Barr virus DNA in gastric carcinoma with lymphoid stroma. Viral Immunol. 1997;10:49–58.PubMedCrossRefGoogle Scholar
  20. 20.
    Zur Hausen A, van Rees BP, van Beek J et al. Epstein-Barr virus in gastric carcinomas and gastric stump carcinomas: a late event in gastric carcinogenesis. J Clin Pathol. 2004;57:487–491.Google Scholar
  21. 21.
    Shukla SK, Prasad KN, Tripathi A, et al. Epstein-Barr virus DNA load and its association with Helicobacter pylori infection in gastroduodenal diseases. Braz J Infect Dis. 2011;15:583–590.PubMedCrossRefGoogle Scholar
  22. 22.
    Minoura-Etoh J, Gotoh K, Sato R, et al. Helicobacter pylori-associated oxidant monochloramine induces reactivation of Epstein-Barr virus (EBV) in gastric epithelial cells latently infected with EBV. J Med Microbiol. 2006;55:905–911.PubMedCrossRefGoogle Scholar
  23. 23.
    Kojima Y, Mogaki M, Takagawa R, et al. A case of lymphoepithelioma-like carcinoma of the colon with ulcerative colitis. J Gastroenterol. 2007;42:181–185.PubMedCrossRefGoogle Scholar
  24. 24.
    Pender MP. Infection of autoreactive B lymphocytes with EBV, causing chronic autoimmune diseases. Trends Immunol. 2003;24:584–588.PubMedCrossRefGoogle Scholar
  25. 25.
    Vaughan JH, Nguyen MD, Valbracht JR, Patrick K, Rhodes GH. Epstein-Barr virus-induced autoimmune responses. II. Immunoglobulin G autoantibodies to mimicking and nonmimicking epitopes. Presence in autoimmune disease. J Clin Invest. 1995;95:1316–1327.PubMedCrossRefGoogle Scholar
  26. 26.
    Harley JB, Harley IT, Guthridge JM, James JA. The curiously suspicious: a role for Epstein-Barr virus in lupus. Lupus. 2006;15:768–777.PubMedCrossRefGoogle Scholar
  27. 27.
    Lunemann JD, Munz C. Epstein-Barr virus and multiple sclerosis. Curr Neurol Neurosci Rep. 2007;7:253–258.PubMedCrossRefGoogle Scholar
  28. 28.
    Swanson-Mungerson M, Longnecker R. Epstein-Barr virus latent membrane protein 2A and autoimmunity. Trends Immunol. 2007;28:213–218.PubMedCrossRefGoogle Scholar
  29. 29.
    Ryan JL, Fan H, Glaser SL, Schichman SA, Raab-Traub N, Gulley ML. Epstein-Barr virus quantization by real-time PCR targeting multiple gene segments: a novel approach to screen for the virus in paraffin-embedded tissue and plasma. J Mol Diagn. 2004;6:378–385.PubMedCrossRefGoogle Scholar
  30. 30.
    Ling PD, Vilchez RA, Keitel WA, et al. Epstein-Barr virus DNA loads in adult human immunodeficiency virus type 1-infected patients receiving highly active antiretroviral therapy. Clin Infect Dis. 2003;37:1244–1249.PubMedCrossRefGoogle Scholar
  31. 31.
    Glaser SL, Gulley ML, Borowitz MJ, et al. Inter- and intra-observer reliability of Epstein-Barr virus detection in Hodgkin lymphoma using histochemical procedures. Leuk Lymphoma. 2004;45:489–497.PubMedCrossRefGoogle Scholar
  32. 32.
    Levine PH, Stemmermann G, Lennette ET, Hildesheim A, Shibata D, Nomura A. Elevated antibody titers to Epstein-Barr virus prior to the diagnosis of Epstein-Barr-virus-associated gastric adenocarcinoma. Int J Cancer.1995;60:642–644.PubMedCrossRefGoogle Scholar
  33. 33.
    van Beek J, van Zur Hausen A, Klein Kranenbarg E, et al. EBV-positive gastric adenocarcinomas: a distinct clinicopathologic entity with a low frequency of lymph node involvement. J Clin Oncol. 2004;22:664–670.PubMedCrossRefGoogle Scholar
  34. 34.
    Oda K, Koda K, Takiguchi N, Nunomura M, Seike K, Miyazaki M. Detection of Epstein-Barr virus in gastric carcinoma cells and surrounding lymphocytes. Gastric Cancer. 2003;6:173–178.PubMedCrossRefGoogle Scholar
  35. 35.
    Luqmani YA, Linjawi SO, Shousha S. Detection of Epstein-Barr virus in gastrectomy specimens. Oncol Rep. 2001;8:995–999.PubMedGoogle Scholar
  36. 36.
    Nakamura S, Ueki T, Yao T, Ueyama T, Tsuneyoshi M. Epstein-Barr virus in gastric carcinoma with lymphoid stroma. Special reference to its detection by the polymerase chain reaction and in situ hybridization in 99 tumors, including a morphologic analysis. Cancer. 1994;73:2239–2249.PubMedCrossRefGoogle Scholar
  37. 37.
    Yuen ST, Chung LP, Leung SY, Luk IS, Chan SY, Ho J. In situ detection of Epstein-Barr virus in gastric and colorectal adenocarcinomas. Am J Surg Pathol. 1994;18:1158–1163.PubMedCrossRefGoogle Scholar
  38. 38.
    Morgan DR, Dominguez RL, Keku TO, et al. Gastric cancer and the high combination prevalence of host cytokine genotypes and Helicobacter pylori in Honduras. Clin Gastroenterol Hepatol. 2006;4:1103–1111.PubMedCrossRefGoogle Scholar
  39. 39.
    Greenberg ER, Anderson GL, Morgan DR, et al. 14-day triple, 5-day concomitant, and 10-day sequential therapies for Helicobacter pylori infection in seven Latin American sites: a randomised trial. Lancet. 2011;378:507–514.PubMedCrossRefGoogle Scholar
  40. 40.
    Cho HJ, Kim JY, Yoo J, Lee SS. Gastric carcinoma with lymphoid stroma: incidence of EBV and Helicobacter pylori infection. Appl Immunohistochem Mol Morphol. 2003;11:149–152.PubMedCrossRefGoogle Scholar
  41. 41.
    Normark S, Nilsson C, Normark BH, Hornef MW. Persistent infection with Helicobacter pylori and the development of gastric cancer. Adv Cancer Res. 2003;90:63–89.PubMedCrossRefGoogle Scholar
  42. 42.
    Wong NA, Herbst H, Herrmann K, et al. Epstein-Barr virus infection in colorectal neoplasms associated with inflammatory bowel disease: detection of the virus in lymphomas but not in adenocarcinomas. J Pathol. 2003;201:312–318.PubMedCrossRefGoogle Scholar
  43. 43.
    Spieker T, Herbst H. Distribution and phenotype of Epstein-Barr virus-infected cells in inflammatory bowel disease. Am J Pathol. 2000;157:51–57.PubMedCrossRefGoogle Scholar
  44. 44.
    Wakefield AJ, Fox JD, Sawyerr AM, et al. Detection of herpesvirus DNA in the large intestine of patients with ulcerative colitis and Crohn’s disease using the nested polymerase chain reaction. J Med Virol. 1992;38:183–190.PubMedCrossRefGoogle Scholar
  45. 45.
    Gehlert T, Devergne O, Niedobitek G. Epstein-Barr virus (EBV) infection and expression of the interleukin-12 family member EBV-induced gene 3 (EBI3) in chronic inflammatory bowel disease. J Med Virol. 2004;73:432–438.PubMedCrossRefGoogle Scholar
  46. 46.
    Ruther U, Nunnensiek C, Muller HA, Bader H, May U, Jipp P. Interferon alpha (IFN alpha 2a) therapy for herpes virus-associated inflammatory bowel disease (ulcerative colitis and Crohn’s disease). Hepatogastroenterology. 1998;45:691–699.PubMedGoogle Scholar
  47. 47.
    Yanai H, Shimizu N, Nagasaki S, Mitani N, Okita K. Epstein-Barr virus infection of the colon with inflammatory bowel disease. Am J Gastroenterol. 1999;94:1582–1586.PubMedCrossRefGoogle Scholar
  48. 48.
    Marszalek A, Marciniak R, Szkaradkiewicz A, et al. Inflammatory bowel disease—is there something new in the immunological background? Folia Histochem Cytobiol. 2011;49:357–362.PubMedGoogle Scholar
  49. 49.
    Maher MM, Nassar MI. Acute cytomegalovirus infection is a risk factor in refractory and complicated inflammatory bowel disease. Dig Dis Sci. 2009;54:2456–2462.PubMedCrossRefGoogle Scholar
  50. 50.
    Sipponen T, Turunen U, Lautenschlager I, Nieminen U, Arola J, Halme L. Human herpesvirus 6 and cytomegalovirus in ileocolonic mucosa in inflammatory bowel disease. Scand J Gastroenterol. 2011;46:1324–1333.PubMedCrossRefGoogle Scholar
  51. 51.
    Baumann MA, Paul CC. Interleukin-5 is an autocrine growth factor for Epstein-Barr virus-transformed B lymphocytes. Blood. 1992;79:1763–1767.PubMedGoogle Scholar
  52. 52.
    Tosato G, Tanner J, Jones KD, Revel M, Pike SE. Identification of interleukin-6 as an autocrine growth factor for Epstein-Barr virus-immortalized B cells. J Virol. 1990;64:3033–3041.PubMedGoogle Scholar
  53. 53.
    Mauser A, Holley-Guthrie E, Zanation A, et al. The Epstein-Barr virus immediate-early protein BZLF1 induces expression of E2F–1 and other proteins involved in cell cycle progression in primary keratinocytes and gastric carcinoma cells. J Virol. 2002;76:12543–12552.PubMedCrossRefGoogle Scholar
  54. 54.
    Israel BF, Kenney SC. Virally targeted therapies for EBV-associated malignancies. Oncogene. 2003;22:5122–5130.PubMedCrossRefGoogle Scholar
  55. 55.
    Kenney S, Theodore E. Woodward award: development of novel, EBV-targeted therapies for EBV-positive tumors. Trans Am Clin Climatol Assoc. 2006;117:55–74.PubMedGoogle Scholar
  56. 56.
    Foxman EF, Iwasaki A. Genome-virome interactions: examining the role of common viral infections in complex disease. Nat Rev Microbiol. 2011;9:254–264.PubMedCrossRefGoogle Scholar
  57. 57.
    Young VB. The intestinal microbiota in health and disease. Curr Opin Gastroenterol. 2012;28:63–69.PubMedCrossRefGoogle Scholar
  58. 58.
    Chai H, Brown RE. Field effect in cancer-an update. Ann Clin Lab Sci. 2009;39:331–337.PubMedGoogle Scholar
  59. 59.
    Meckes DG Jr, Raab-Traub N. Microvesicles and viral infection. J Virol. 2011;85:12844–12854.PubMedCrossRefGoogle Scholar
  60. 60.
    Pegtel DM, van de Garde MD, Middeldorp JM. Viral miRNAs exploiting the endosomal-exosomal pathway for intercellular cross-talk and immune evasion. Biochim Biophys Acta. 1809;2011:715–721.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Julie L. Ryan
    • 1
  • You-Jun Shen
    • 2
  • Douglas R. Morgan
    • 3
  • Leigh B. Thorne
    • 4
  • Shannon C. Kenney
    • 5
  • Ricardo L. Dominguez
    • 6
  • Margaret L. Gulley
    • 4
  1. 1.Department of Dermatology and Radiation OncologyUniversity of Rochester Medical CenterRochesterUSA
  2. 2.Sentara Virginia Beach General HospitalVirginia BeachUSA
  3. 3.Gastroenterology and Hepatology Division, Department of MedicineUniversity of North CarolinaChapel HillUSA
  4. 4.Department of Pathology and Laboratory Medicine, Lineberger Comprehensive Cancer CenterUniversity of North CarolinaChapel HillUSA
  5. 5.Departments of Medicine and OncologyUniversity of WisconsinMadisonUSA
  6. 6.Department of GastroenterologyWestern Regional HospitalSanta Rosa de CopanHonduras

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