Epstein-Barr Virus Strains With Latent Membrane Protein-1 Deletions: Prevalence in the Italian Population and High Association With Human Immunodeficiency Virus–Related Hodgkin's Disease

Abstract

Six Epstein-Barr virus (EBV)-related lymphoproliferative disorders were investigated to verify whether the EBV strain harbored by neoplastic cells had the same EBNA-2 and latent membrane protein-1 (LMP-1) DNA sequences of the virus carried by normal lymphocytes of the same patients. Within each case, the analysis of neoplastic lymph nodes, reactive lymphadenopathies, and/or EBV+ spontaneous lymphoblastoid cell lines gave concordant results with respect to type-specific EBNA-2 region and LMP-1 gene. In particular, five cases showed the same deletion in the 3′ end of the LMP-1 gene in both normal and neoplastic cells. We also determined the prevalence of LMP-1 deletions in a large series of normal peripheral blood mononucleated cells (PBMCs) from Italian individuals. The analysis showed that 50% (9 of 18) of PBMCs from human immunodeficiency virus (HIV)-seronegative donors carried a 30-bp deletion in the C-terminal portion of the LMP-1 gene, whereas a nondeleted fragment was amplified in about 44% (8 of 18) of the cases. Only one sample (5.6%) showed the amplification of a full-length LMP-1 band together with a deleted fragment. Similarly, PBMCs from HIV-infected patients showed an almost equivalent prevalence of full-length (17 of 37, 46%) and deleted (16 of 37, 43.2%) LMP-1 fragments, whereas about 11% of samples (4 of 37) showed evidence of double infections. Of note, deletions in the LMP-1 gene were detected with similar prevalence values in EBV+ Hodgkin's disease (HD) (13 of 30, 43.3%) and non-Hodgkin's lymphoma (NHL) (2 of 5, 40%) cases from HIV-seronegative patients and in HIV-related, EBV+ NHLs (4 of 7, 57.1%). Conversely, a 30-bp LMP-1 deletion was found in 10 of 12 HIV-associated HD cases (83%), a prevalence significantly higher than that detected in HIV-unrelated HD (P = .01). These findings indicate that: (1) the same EBV strain carrying LMP-1 deletions is harbored by normal and neoplastic cells of patients with EBV+ disorders, ruling out that these mutations might result from immunoselection phenomena; (2) in the Italian population, the prevalence of LMP-1 deletion mutants is comparable to that of EBV strains with full-length LMP-1; (3) HIV-induced immunosuppression is not associated with an increased prevalence of LMP-1 deletions in PBMCs; and (4) HIV-related HD cases, but not those of HIV-seronegative Italian patients, are closely correlated with the presence of LMP-1 deletions, suggesting that infection with these strains may increase the risk of developing HD in the HIV setting.

Epstein–Barr virus strains and variations: Geographic or disease‐specific variants?

“…Similar results were found by other authors [Chang et al, 2009;Bobek et al, 2010;Banko et al, 2012;Gurtsevitch et al, 2013;Senyuta et al, 2014]. Other studies have also associated the del-LMP-1 variant to the development of IM [Berger et al, 1997;Ai et al, 2012;Lorenzetti et al, 2012] and several malignancies, including Hodgkin's Lymphoma (HL) [Zhou et al, 2001;Guiretti et al, 2007;Ai et al, 2012], human immunodeficiency virus-related HL [Dolcetti et al, 1997;Garc ıa-Cos ıo et al, 2008], peripheral T/natural killer (NK)-cell lymphoma [Sandvej et al, 1994;Chiang et al, 1999], and other non-Hodgkin's diseases [Kingma et al, 1996;Mansoor et al, 1997]. The literature has also shown that EBV-associated gastric cancer (EBVaGC) has been strongly associated with the del-LMP1 variant [Chen et al, 2010BenAyed-Guerfali et al, 2011;Cheng et al, 2015].…”

Epstein‐barr virus (EBV) in healthy carriers: Distribution of genotypes and 30 bp deletion in latent membrane protein‐1 (LMP‐1) oncogene

“…Studies of normal or reactive lymphoid tissues from Brazil and Mexico found the del-LMP-1 variant in 59% and 73%, respectively [Chen et al, 1996;Dirnhofer et al, 1999]. Results in PBMCs from healthy Italian donors showed similar proportions of the two variants (55% of non-deleted and 44% of deleted LMP-1) [Dolcetti et al, 1997]. A predominance of co-infections (deleted and non-deleted variants) in multiple epithelial and lymphoid tissues (93%) was reported in healthy carriers in Hong Kong [Srivastava et al, 2000].…”

Human herpesvirus 7, Epstein–Barr virus and human cytomegalovirus in periodontal tissues of periodontally diseased and healthy subjects

“…The exons 1-4 of this region encode the IE1 transactivator which appears in infected cells 1 h post infection and has an important function in the regulation of HCMV genes (Pignatelli et al 2004). EBV was searched using primers specific for type-1 and type-2 EBV LMP-1 region (Dolcetti et al 1997) (Table 1), which encode an integral membrane protein of 386 amino acids able to exert pleiotropic effects essential for EBV-induced B-cell transformation (Eliopoulos & Rickinson 1998). This region is highly heterogeneous across diverse isolates, showing clustered point mutations, insertions and in frame 30 or 69 bp deletions (Hudson et al 1985, Weiss et al 1987, Carbone et al 1993.…”

“…This region is highly heterogeneous across diverse isolates, showing clustered point mutations, insertions and in frame 30 or 69 bp deletions (Hudson et al 1985, Weiss et al 1987, Carbone et al 1993. Primers were chosen within regions of the LMP-1 carboxy terminus (Dolcetti et al 1997).…”