Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contact Us Login 
An Official Publication of the Indian Association of Oral and Maxillofacial Pathologists

  Table of Contents    
Year : 2021  |  Volume : 25  |  Issue : 4  |  Page : 46-50

Epstein–Barr virus-positive ulcer in the oral cavity

1 Oral and Maxilloficial Pathology Area, ENES-Leon, National Autonomous University of Mexico, Mexico City, Mexico
2 Department of Oral Medicine and Pathology, Postgraduate Division, Dental School, National Autonomous University of Mexico, Mexico City, Mexico

Date of Submission18-Jan-2019
Date of Decision05-Aug-2019
Date of Acceptance28-Sep-2020
Date of Web Publication19-Mar-2021

Correspondence Address:
Javier Portilla-Robertson
Department of Oral Pathology, Graduate Dental School, National Autonomous Mexico University, Av. Universidad # 3000, Mexico City
Login to access the Email id

Source of Support: None, Conflict of Interest: None

DOI: 10.4103/jomfp.JOMFP_26_19

Rights and Permissions



Epstein–Barr virus-positive ulcer (EBV + U) is a recently reported B cell lymphoproliferative disorder in the oral cavity, oropharynx, gastrointestinal tract and skin, principally in immunosuppressed patients. A 53-year-old female patient with rheumatoid arthritis treated with methotrexate, presenting ulcers of unknown duration on the dorsum and the lateral left border of the tongue. Excisional biopsy, histopathological analysis and histochemical stains for syphilis (Warthin–Starry), mycotic diseases (Grocott silver methenamine), tuberculosis (Ziehl–Neelsen), immunohistochemistry tests for herpesvirus type 8 (CMV), EBV (LMP-1) and DNA extraction for polymerase chain reaction (PCR) assay to CMV, EBV and herpes simplex virus-1 were performed. Posterior to PCR assay, the final diagnosis was EBV + U in the oral cavity. Acyclovir® was prescribed, showing clinical improvement. A case of EBV + U with clinical characteristics similar to other lesions or conditions has been reported. Special assays are necessary for an accurate diagnosis and treatment.

Keywords: Acyclovir, chronic ulcers, Epstein–Barr virus, methotrexate

How to cite this article:
Ramos-Baena JD, Jacinto-Alemán LF, Leyva-Huerta ER, Portilla-Robertson J. Epstein–Barr virus-positive ulcer in the oral cavity. J Oral Maxillofac Pathol 2021;25, Suppl S1:46-50

How to cite this URL:
Ramos-Baena JD, Jacinto-Alemán LF, Leyva-Huerta ER, Portilla-Robertson J. Epstein–Barr virus-positive ulcer in the oral cavity. J Oral Maxillofac Pathol [serial online] 2021 [cited 2022 Aug 18];25, Suppl S1:46-50. Available from: https://www.jomfp.in/text.asp?2021/25/4/46/311546

   Introduction Top

Epstein–Barr virus (EBV) is present in 90% of the population. It has a predilection for B cells, in which it causes proliferation and transformation.[1] Its viral latency is established in memory B cells after primary infection, which occurs mainly during childhood and adolescence.[2] The cell-mediated immune system keeps B cells infected by EBV (B-EBV +) under viral latency. Therefore, drug-related immunosuppression allows B-EBV + to proliferate resulting in a wide range of lymphoproliferative disorders immunodeficiency associated. The above is observed in patients with lymphoproliferative diseases associated with primary immune disorders or posttransplanting (iatrogenic immunodeficiency).[3]

EBV-positive ulcer (EBV+U) is a newly described entity consisting of well-defined and isolated lesions that occur in the skin, oral cavity or gastrointestinal tract. the morphological characteristics of EBV+U may vary, but usually, an ulcer with mixed lymphoplasmacytic infiltrates on a background of necrotic tissue is found. Cells within the infiltrate consist of small T lymphocytes, large B lymphocytes and often Reed–Sternberg-like cells that coexpress B cell and CD30 antigens.[4] EBV+U occurs predominantly in patients with age-related immunosuppression or iatrogenic immunosuppression by autoimmune disease treatment.[5],[6]

EBV+U could be developed in the oral mucosa since the oral cavity has the highest concentration of infected B cells in Waldeyer's ring. This alteration was first described in 2010 by Dojcinov et al.[3] in a study that included 26 patients that presented ulcers in the oral cavity, skin and colon, related to some type of immunosuppression. They proposed the EBV+U as a new clinicopathological entity with histopathological characteristics similar to Hodgkin's lymphoma but self-limited, which generally responds well to the administration of conservative treatments, such as the regulation of immunosuppressive drugs.

   Case Report Top

A 53-year-old female patient was referred to her general dentist, to our Oral Medicine Clinic of the Postgraduate and Research Division, Faculty of Dentistry National University of Mexico (UNAM). The patient complained of ulcers on the dorsum and left border of the tongue. On interrogation, she mentioned suffering rheumatoid arthritis with 30 months of evolution, treated with methotrexate and prednisone as well as hypertension for 2 years under metoprolol medication. Intraoral examination confirms the presence of an ulcer on the left border of her tongue, with well-defined and indurated borders, about 0.7 cm in length and another ulcer on the dorsum of the tongue also well-defined and indurated borders, measuring 1.8 cm × 0.5 cm × 0.5 cm [Figure 1]a and [Figure 1]b. Both ulcers caused painful symptoms. The patient reported 4 months of evolution, increasing in size and symptoms. The patient referred the previous prescription of diverse pharmacotherapy indicated by general dentists such as amoxicillin, ketoconazole and nystatin without improvement.
Figure 1: (a) Ulcer in the tongue dorsum (b) Ulcer in the left border of the tongue, (c) Macroscopic analysis of both specimen labeled as A and B, (d) photomicrography at ×100 magnifications that show the discontinuity of epithelium with inflammatory infiltrate, (e) photomicrography at ×400 magnifications showing cytopathic changes suggestive of viral infection in epithelial cells and (f) photomicrography at ×400 magnifications showing the high presence of lymphoplasmacytic infiltrate underlying conjunctive tissue

Click here to view

Our differential clinical diagnosis included mycosis, traumatic ulcerative granuloma with stromal eosinophils or oral squamous cell carcinoma. An excisional biopsy was performed; hemostasis with electrocautery was fulfilled in both lesions. Specimens were fixed in 10% formalin and send to our histopathological diagnostic service in our institution.

The specimens were labeled as follows: (A) single oval-shaped specimen of soft tissue, measuring 2.2 cm × 1.5 cm × 1 cm, irregular surface with a poorly delimited ulcer, irregular edges, firm consistency, dark and light brown color toward the center, edges were inked and representative sections were included. (B) A single specimen measuring 1 cm × 0.7 cm × 0.5 cm, with an oval shape, ulcerated, firm consistency, dark brown color and the edges were inked [Figure 1]c.

The histological sections showed loss of continuity of the epithelium replaced by fibrin, the connective tissue presented abundant mixed inflammatory infiltrate predominantly lymphoplasmacytic, some cells exhibited pleomorphism. In some areas, biological material that resembled a foreign body reaction with the presence of epithelioid cells, histiocytes and multinucleated giant cells was also observed, as well as cytopathic effect in the periphery of the ulcerated epithelium [Figure 1]d, [Figure 1]e, [Figure 1]f.

The patient's rheumatoid arthritis condition impaired to assist to her postoperative checkup and removal of suture appointment. She attended to her private general dentist, which was contacted and reported that the postsurgical zone was observed in unfavorable conditions for both lesions.

According to microscopic findings, a diagnosis of a nonspecific chronic ulcer with granulomatous foreign body reaction was emitted; however, the special stains to detect the presence of fungi (Grocott methenamine silver), for Mycobacterium tuberculosis (Ziehl–Neelsen) and for Treponema pallidum (Warthin–Starry), were negative [Figure 2].
Figure 2: Photomicrographs at ×100 magnifications of special stains, (a) Grocott methenamine silver, (b) Staining with Ziehl–Neelsen and, (c) Staining with Warthin–Starry. Negative results were obtained

Click here to view

In relation to the cytopathic changes, treatment with 200 mg acyclovir 5 times daily for 3 weeks and clinical evaluation at 2 weeks was scheduled. The patient again failed to her evaluation appointment, but she provided photographs of her clinical progress treatment, showing resolution of the ulcer located on the lateral left border of the tongue and improvement in ulcer located on the dorsum of the tongue, with a decrease in size and depth.

To validate the response to herpes virus treatment, endpoint polymerase chain reaction (PCR) assay was performed, for which DNA extraction was done from 50 μm thick slice, using the ReliaPrep™ FFPE gDNA Miniprep system (Promega, MA, USA); the quality and amounts of DNA were determinate through NanoDrop 2000 spectrophotometer (Thermo Fisher, Wilmington, DE, USA).

A total of 100 ng of DNA was amplified with the PCR master mix kit (Promega, Madison, WI). Primers used to identify LMP1 EBV, CMVUL44 and HVS UL56; listed in [Table 1], obtained from Eurofin mwg/operon (Louisville, KY). Glyceraldehyde-3-phosphate dehydrogenase was employed as internal control and normalization reference. Amplifications were carried out in a MaxyGene II thermal cycler (Axygen, NY) carrying out 40 cycles. Amplification products were resolved on 2% agarose gels in TAE buffer (Amresco, Solon, OH) at 100 mV. The gels were stained with 0.05 μg/ml ethidium bromide (Promega, Madison, WI) and visualized in a gel documentation system (Axygen, NY), to determine their expression by optical density in GelQuantNet (BiochemLab Solutions, UCSF, CA, USA). Amplification assay was performed in triplicate obtaining predominant positivity for EBV, with the presence of CMV and herpes simplex virus (HSV) [Figure 3].
Table 1: Primers' sequence for polymerase chain reaction

Click here to view
Figure 3: Viral detection by polymerase chain reaction analysis. (a) Media expression of VHS, CMV and Epstein–Barr virus; Glyceraldehyde-3-phosphate dehydrogenase-normalized; and (b) polymerase chain reaction amplification products resolved on agarose 2% gel. Predominant expression in Epstein–Barr virus-A sample was observed

Click here to view

Two weeks later, the patient attended to our clinic, commenting that she did not finish the treatment as indicated since she felt improvement during the 1st day. The lesion in the dorsum tongue was still present as well as painful symptoms. It was decided to resume the therapy with acyclovir as it was previously prescribed.

The patient failed again to her appointment because of a hip fracture, but she provided clinical photographs showing the resolution of the other lesion, without symptomatology [Figure 4].
Figure 4: Follow-up 7 days after starting treatment. (a) Partial healing of the dorsum tongue ulcer and (b) total healing of the left lateral ulcer of the tongue

Click here to view

   Discussion Top

EBV+U is a lymphoproliferative disease described for the first time in 2010 by Dojcinov et al.,[3] in which a series of 26 cases were reported showing mucocutaneous ulcers that slowly developed induration, involving the oropharyngeal mucosa, gastrointestinal tract or skin, with characteristic histological and immunophenotypic characteristics of lymphoproliferative disease associated with EBV, related to different forms of immunosuppression.[7],[8],[9],[10],[11]

Ulcers in the oral cavity may occur in oral mucosa, gingiva or tongue, as in our case; it can be related to chronic inflammation by local trauma, resulting in the formation of an ulcer and localized proliferation of B cells infected with EBV.[12]

The actual incidence of EBV+U is unknown. In a recent literature review, Roberts et al., in 2016 include 51 cases, in which the most frequent causes were immunosuppression and the most common drugs used were as follows: methotrexate, azathioprine and cyclosporine in 56% of the cases; combined with advanced age in 40%. Clinically, EBV+U usually manifests as a single lesion in 84% of cases as mucosal or cutaneous ulcer, well-circumscribed, without lymph node involvement. The oropharynx was the most common site affected, followed by skin.[6]

The main histological features include a polymorphic inflammatory infiltrate with lymphocytes, histiocytes, eosinophils and plasma cells mixed with scattered atypical B cells, some with Reed–Sternberg-type morphology. These atypical lymphocytes Reed–Sternberg type are positive for CD30, CD15, MUM1, PAX5 and OCT-2 and mostly negative for CD20, CD79a, CD45, CD3 and Bcl-6. In relation to auxiliary assays for diagnosis, the in situ hybridization for EBV-encoded small RNA (EBER) oligonucleotides is positive by definition. In some cases, large B cells that infiltrated the arteries are associated with thrombosis and necrosis.[5] In addition, heavy chains of monoclonal immunoglobulin and T cell receptor (TCR) rearrangement have been observed in up to 40% of cases, suggesting a clonal proliferation of B cells driven by EBV.[3]

This is the first time that PCR assay was used for EBV + U diagnosis and this assay proved results as an extremely useful strategy to determine the specific genotype of the virus.

Their applications to lymphoproliferative disorders associated with EBV today have been restricted to determine the clonality of the TCR and immunoglobulins, nevertheless, with this finding, their scope could be extended.[13]

The EBV + U differential diagnosis includes other lymphoproliferative disorders and neoplasms, especially Hodgkin's lymphoma and diffuse B-cell lymphoma.[5] In our case, the clinical diagnosis was unclear, since we considered a squamous cell carcinoma, syphilis, tuberculosis, fungal infection or lymphoma; however, the anatomical location of the lesion, the histopathological findings and the PCR assay favored the final diagnosis of EBV+U.

Several personal problems of the patient, such as her general systemic health complications by arthritis and hypertension and her lack of attachment to guidelines indicated hampered the diagnosis, treatment and follow-up of this case. The therapy used was acyclovir, an antiviral HSV drug, which promoted clinical improvement, validated by our PCR results. Acyclovir can inhibit lytic infection by the herpes virus when it is phosphorylated by viral kinases and incorporated into viral DNA.[14],[15] To date, there are no reports of this therapy or management of these alterations, so considering the above, this could be an additional measure for EBV+U cases that occur during pharmacological immunosuppression.[16],[17],[18] In our case, an important limiting was the patient lack to therapeutic guidelines suggested and despite that improvement was observed; however, more clinical studies are needed to corroborate our findings.

Declaration of patient consent

The authors certify that they have obtained all appropriate patient consent forms. In the form the patient(s) has/have given his/her/their consent for his/her/their images and other clinical information to be reported in the journal. The patients understand that their names and initials will not be published and due efforts will be made to conceal their identity, but anonymity cannot be guaranteed.

Financial support and sponsorship

UNAM PAPIME 209016 and PAPIIT 223414.

Conflicts of interest

There are no conflicts of interest.

   References Top

Onafowokan O, Rodgers W, Gruber E. Epstein-Barr virus-positive mucocutaneous ulcer of the oral cavity in a patient who had had a renal transplant. Br J Oral Maxillofac Surg 2018;56:626-8.  Back to cited text no. 1
Juan A, Lobatón T, Tapia G, Mañosa M, Cabré E, Domènech E. Epstein-Barr virus-positive mucocutaneous ulcer in Crohn's disease. A condition to consider in immunosuppressed IBD patients. Dig Liver Dis 2017;49:934-7.  Back to cited text no. 2
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;34:405-17.  Back to cited text no. 3
Hart M, Thakral B, Yohe S, Balfour HH Jr, Singh C, Spears M, et al. EBV-positive mucocutaneous ulcer in organ transplant recipients: A localized indolent posttransplant lymphoproliferative disorder. Am J Surg Pathol 2014;38:1522-9.  Back to cited text no. 4
McGinness JL, Spicknall KE, Mutasim DF. Azathioprine-induced EBV-positive mucocutaneous ulcer. J Cutan Pathol 2012;39:377-81.  Back to cited text no. 5
Roberts TK, Chen X, Liao JJ. Diagnostic and therapeutic challenges of EBV-positive mucocutaneous ulcer: A case report and systematic review of the literature. Exp Hematol Oncol 2015;5:13.  Back to cited text no. 6
Au JK, Said JW, Sepahdari AR, St John MA. Head and neck Epstein-Barr virus mucocutaneous ulcer: Case report and literature review. Laryngoscope 2016;126:2500-4.  Back to cited text no. 7
Furukawa S, Oobu K, Moriyama M, Kawano S, Sako S, Hayashida JN, et al. Oral Methotrexate-related lymphoproliferative disease presenting with severe osteonecrosis of the jaw: A case report and literature review. Intern Med 2018;57:575-81.  Back to cited text no. 8
Hujoel IA, Rubio-Tapia A, Dao LN, Porrata LF, Kane SV. Epstein-Barr virus-positive mucocutaneous ulcer in an immunosuppressed patient. ACG Case Rep J 2018;5:e32.  Back to cited text no. 9
Yamakawa N, Fujimoto M, Kawabata D, Terao C, Nishikori M, Nakashima R, et al. A clinical, pathological, and genetic characterization of methotrexate-associated lymphoproliferative disorders. J Rheumatol 2014;41:293-9.  Back to cited text no. 10
Kalantzis A, Marshman Z, Falconer DT, Morgan PR, Odell EW. Oral effects of low-dose methotrexate treatment. Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2005;100:52-62.  Back to cited text no. 11
Kikuchi K, Miyazaki Y, Tanaka A, Shigematu H, Kojima M, Sakashita H, et al. Methotrexate-related Epstein-Barr virus (EBV)-associated lymphoproliferative disorder--so-called “Hodgkin-like lesion”--of the oral cavity in a patient with rheumatoid arthritis. Head Neck Pathol 2010;4:305-11.  Back to cited text no. 12
Dojcinov SD, Venkataraman G, Pittaluga S, Wlodarska I, Schrager JA, Raffeld M, et al. Age-related EBV-associated lymphoproliferative disorders in the Western population: A spectrum of reactive lymphoid hyperplasia and lymphoma. Blood 2011;117:4726-35.  Back to cited text no. 13
Kutok JL, Wang F. Spectrum of Epstein-Barr virus-associated diseases. Annu Rev Pathol 2006;1:375-404.  Back to cited text no. 14
Khan N, Hislop A, Gudgeon N, Cobbold M, Khanna R, Nayak L, et al. Herpesvirus-specific CD8 T cell immunity in old age: Cytomegalovirus impairs the response to a coresident EBV infection. J Immunol 2004;173:7481-9.  Back to cited text no. 15
Attard AA, Praveen P, Dunn PJ, James GJ. Epstein-Barr virus-positive mucocutaneous ulcer of the oral cavity: the importance of having a detailed clinical history to reach a correct diagnosis. Oral Surg Oral Med Oral Pathol Oral Radiol. 2012;114:e37-9.  Back to cited text no. 16
Kojima M, Kashimura M, Itoh H, Noro A, Akikusa B, Iijima M, et. al. Epstein-Barr virus-related reactive lymphoproliferative disorders in middle-aged or elderly patients presenting with atypical features. A clinicopathological study of six cases. Pathol Res Pract. 2007;203:587-91.  Back to cited text no. 17
Asano N, Yamamoto K, Tamaru J, Oyama T, Ishida F, Ohshima K, et al. Age-related Epstein-Barr virus (EBV)-associated B-cell lymphoproliferative disorders: comparison with EBV-positive classic Hodgkin lymphoma in elderly patients. Blood. 2009;113:2629-36.  Back to cited text no. 18


  [Figure 1], [Figure 2], [Figure 3], [Figure 4]

  [Table 1]


Print this article  Email this article


    Similar in PUBMED
    Search Pubmed for
    Search in Google Scholar for
  Related articles
    Article in PDF (1,826 KB)
    Citation Manager
    Access Statistics
    Reader Comments
    Email Alert *
    Add to My List *
* Registration required (free)  

   Case Report
    Article Figures
    Article Tables

 Article Access Statistics
    PDF Downloaded172    
    Comments [Add]    

Recommend this journal

Journal of Oral and Maxillofacial Pathology | Published by Wolters Kluwer - Medknow
Online since 15th Aug, 2007