The association of Epstein-Barr Virus infection with ...

Original Article

The association of Epstein-Barr Virus infection with multiple myeloma Mohammad Hadi Sadeghian, Hossein Ayatollahi, Mohammad Reza Keramati1, Bahram Memar2, Saeed Amel Jamedar3, Maryam Mohammadnia Avval, Maryam Sheikhi, Gohar Shaghayegh Department of Hematology and Blood Bank, Faculty of Medicine, 1Neonatal Research Center, Imam Reza Hospital, MUMS, 2 Department of Pathology, 3Department of Microbiology, Mashhad University of Medical Sciences, Mashhad, Iran. Address for correspondence: Dr. Hossein Ayatollahi, Department of Hematology and Blood Bank, Endoscopic & Minimally Invasive Surgery Research Center, Ghaem Medical Center, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran. E-mail: [email protected]

ABSTRACT

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Background: Multiple myeloma is a conventional term for clonal proliferation of plasma cells. It is suggested that viruses may have a significant role in the pathogenesis of multiple myeloma. Aims: The aim of this study was to evaluate whether there is an association between multiple myeloma and Epstein-Barr Virus (EBV) by the polymerase chain reaction (PCR) method. Materials and Methods: This case-control study was performed on 60 paraffin-embedded bone marrow biopsies (30 multiple myeloma and 30 normal bone marrow specimens) in the molecular pathology section of our hospital. The patients and control groups were matched according to gender and age. Several sections were cut from each paraffin blocks, and then the deoxyribonucleic acid (DNA) was extracted by the non-heating extraction method. In the next step, PCR was carried out for detection of EBV genome and finally its products were analyzed by electrophoresis. Results: DNA of EBV was detected in 10 patients of the case group (5 males and 5 females) and 3 subjects (2 males and 1 female) of the control group. The Pearson chi-square test showed significant difference between case and control group (P=0.03) for detection of the EBV genome. In myeloma patients, the mean white blood cell count was 9.05 + 4.02 and 5.20 + 2.02 × 109/L in EBV positive and negative groups, respectively and a significant difference count was seen for the WBC count. Recommendations: Our results show an association between multiple myeloma and EBV infection. KEY WORDS: Epstein-Barr Virus, multiple myeloma, polymerase chain reaction

INTRODUCTION Multiple myeloma (MM) is a malignant disorder of plasma cells, primarily occurring in the bone marrow. This neoplastic lesion is diagnosed based on pathology, radiologic, and clinical findings, including anemia, hypercalcemia, renal insufficiency, high serum immunoglobulin, and radiologic evidence of lytic bone lesions. The term of “plasma cell myeloma” is entered in the 2008 WHO (World Health Organization) classification of malignant B-cell lymphoma instead of MM.[1] Near to 1% of all neoplasm and 10% of hematologic malignancies in the United States are Multiple myeloma.[2] MM also accounts for about 20% of the deaths caused by hematologic neoplasms.[3] There are some data suggesting the possible potential role of several infectious agents in the pathogenesis of MM. Montella et al. [4] implicated the association of hepatitis C virus infection with MM and Elira Dokekias et al. [5] reported plasma cell myeloma in human immunodeficiency virus (HIV)-infected patients. Although some authors have found a relationship between human herpes virus 8 (HHV-8), a member of the herpes virus family and MM [6-8] but this finding has not been confirmed by others.[9-11] 720

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Website: www.ijpmonline.org PMID: xxxxxxxxx (when available) DOI: 10.4103/0377-4929.91504 Quick Response Code:

Epstein-Barr Virus (EBV) is another member of the herpesvirus family and causes infectious mononucleosis. It is well documented that this virus is associated with a number of malignancies such as nasopharyngeal carcinoma, a subset of Hodgkin lymphoma, the African (endemic) form of Burkitt lymphoma, B-cell lymphomas in immunosuppressed individuals and rare forms of T-cell lymphomas. [11] The role of EBV in pathogenesis of some form of lymphoprolifrative disorders is so important that even the name of this virus has been entered in classification of lymphoma and some EBV-related neoplasms (e.g., EBV+ diffuse large B-cell lymphoma of the elderly, pediatric follicular lymphoma, and EBV+ T-cell lymphoproliferative diseases of childhood) are highlighted in the new (2008) WHO classification. [13] There are few reports about detection of EBV genome in MM patients[13-16] but this finding has not been confirmed by others.[16-18] The aim of this study was to investigate whether there are differences between two groups of patients with MM and the

MICROBIOLOGY - 54(4), OCTOBER-DECEMBER 2011

Sadeghian, et al.: Epstein - Barr Virus and multiple myeloma

control group for the presence of EBV deoxyribonucleic acid (DNA) in bone marrow tissue by the polymerase chain reaction (PCR) method. MATERIAL AND METHODS This case-control study financially was supported and ethically approved by the research vice chancellor of Mashhad University of Medical Sciences. Pathology Diagnosis It was performed on 60 formalin-fixed paraffin embedded (FFPE) bone marrow biopsies in molecular pathology laboratory, Ghaem teaching Hospital, Mashhad, Iran. Subjects Our materials included 30 FFPE of MM patients who had positive clinical and radiological finding of MM and also had more than 30% plasma cells in the bone marrow (patient group), and 30 FFPE of normal subjects had not increase plasma cells (control group). Control specimens were obtained from patients with lymphoma that had normal bone marrow morphology and they were matched with patients group for age and sex. The bone marrow biopsy in patients with lymphoma had been performed for determination of staging. Archived slides of two groups were reviewed by two pathologists for confirmation of diagnosis and tissue adequacy for extracting of DNA. We also reviewed medical history and previous laboratory reports of patients. We did not find any history of immunodeficiency diseases, transplantation, or immunosuppressive therapy in our patients. DNA Extraction Five to seven 20-μm-thick sections were cut from each FFPE specimens under sterile conditions, and then the DNA was extracted by using proteinase K and the non-heating DNA extraction method [19] DNA concentration was determined by using the Thermo Scientific NanoDrop 2000 Spectrophotometer and specimens with low DNA content (90%) in patients with Hodgkin lymphoma by the immunohistochemistry method in Northeast Iran. Our study was done on 30 patients with MM. This sample size may be too small for final decision; therefore, future study with a larger number of case groups is helpful to decrease the probable sampling error. Furthermore, complementary studies by using in situ hybridization or immunohistochemistry methods are needed to detect direct evidence of EBV in bone marrow tissue.

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CONCLUSION We find a statistically significant relationship between multiple myeloma and detection of EBV DNA in bone marrow tissues by the PCR method but because of the high prevalence of EBV infection in the control group, a similar study with greater number of patients can be helpful for the final decision. ACKNOWLEDGEMENT

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This study was the result of a MD thesis and financially supported by the research vice chancellor of Mashhad University of Medical Sciences. The authors would like to thank Dr. Mohammad Khajeh Daluei for his invaluable recommendations and data analysis. INDIAN JOURNAL

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How to cite this article: Sadeghian MH, Ayatollahi H, Keramati MR, Memar B, Jamedar SA, Avval MM, Sheikhi M, Shaghayegh G. The association of Epstein-Barr Virus infection with multiple myeloma. Indian J Pathol Microbiol 2011;54:720-4. Source of Support: The research vice chancellor of Mashhad University of Medical Sciences, Conflict of Interest: None declared.

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