Epstein-Barr Virus Polymerase Chain Reaction Research Articles

Subclinical EBV and CMV Reactivations Commonly Occur Following Antibody-Based Immunosuppressive Regimens for Patients with Severe Aplastic Anemia.

Although reactivation of Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) during periods of immune suppression following solid organ and allogeneic hematopoietic stem cell transplantation (HSCT) can cause fatal complications, the natural history of viral reactivation and the potential for serious complications following antibody-based immunosuppressive (I/S) treatments for bone marrow failure syndromes is not defined. We monitored blood for EBV and CMV reactivation by polymerase chain reaction (PCR) weekly in 39 consecutive patients (total of 44 I/S courses) with aplastic anemia (AA). During the observation period of 3–6 months per patient, none developed clinical features of EBV or CMV reactivation, nor received antiviral therapy. Four regimens were studied: (1) HC: horse ATG + cyclosporine (CsA) (18 pts); (2) HCS: horse ATG + CsA + sirolimus (15 pts); (3) RC: rabbit ATG (Thymoglobulin®) + CsA (5 pts); and (4) CP: alemtuzumab (Campath®) (6 pts). Overall EBV reactivation occurred in 32/39 (82%) seropositive patients at a median of 14 days in the HC arm, 11 days in the HCS arm, 15 days in the RC arm and 6 days in the CP arm from starting I/S. The median peak EBV copies for HC recipients was 4,340 (0–54,250) copies/106 mononuclear cells (MNCs); for HCS 1,420 (0–466,000) copies/106 MNCs; for RC 370,000 (97,000–1,025,000) copies/106 MNCs, and for CP 965 copies/106 MNCs (0–655,000) (P= 0.026). The median duration of PCR positivity for EBV was higher in the RC group (18 weeks) compared to HC (6 weeks), HCS (4 weeks) and CP (1 week) (P=0.002). Higher peak values were associated with a longer duration of viremia following initial I/S (P<0.001)(Figure). CMV reactivation occurred only in 12/29 (41%) seropositive patients, 10 CMV seronegative patients showed no reactivation by PCR. The EBV reactivations in the alemtuzumab arm were less frequent and short lived, presumably because the antibody eliminates the B cell population which is the source for reactivating virus. A more prolonged lymphocyte depletion was seen with RC and CP treated patients when compared to HC and HCS. Our results indicate that subclinical reactivation of both EBV and CMV is common in patients with AA receiving I/S and that different regimens are associated with different intensity of I/S as measured by viral load and lymphocyte count. Our data suggests that higher and longer EBV reactivations are seen following rabbit ATG compared to horse ATG or alemtuzumab, and for the whole cohort, the duration of positivity for EBV PCR correlated to the initial degree of EBV reactivation. While all reactivations were subclinical, our findings suggest that viral reactivation should be monitored when developing new and more potent I/S regimens for treatment of hematologic and non-hematologic disorders as the threshold to the development of CMV or EBV disease is not well understood. [Display omitted]

The impact of monitoring Epstein–Barr virus PCR in paediatric bone marrow transplant patients: Can it successfully predict outcome and guide intervention?

Epstein-Barr virus (EBV) associated lymphoproliferative disease is a complication of haemopoietic stem cell transplantation (HSCT). In certain groups (unrelated and mismatched donor transplants, T-cell depleted) the risk may be as high as 25% with significant morbidity and mortality. Strategies to predict the impending development of this disorder and allow early intervention have therefore assumed importance. We routinely screen the peripheral blood of all recipients of allogeneic HSCT to detect EBV DNA by quantitative polymerase chain reaction (PCR) technology and report here how this correlates with clinical disease and management. Data on 28 successive patients who underwent HSCT at our institution were reviewed. The relationship between EBV reactivation demonstrated by quantitative PCR and development of post transplant lymphoproliferative disease (PTLD) was determined. EBV reactivation occurred in 68% of patients, however only 7% developed clinical PTLD. Patients with high level reactivation (n = 9) had more frequent episodes of reactivation and all patients who progressed to overt PTLD were found in this group. In contrast none of those patients with low level reactivation (n = 10) or persistently negative results (n = 9) showed any signs of clinical disease. Anti-CD20 monoclonal antibody (Rituximab) therapy was instigated in both cases of proven PTLD and three cases of high level reactivation with successful outcomes. Response to treatment was associated with a prompt decline in viral copy number. Our results indicate that EBV reactivation is a common occurrence in the paediatric allogeneic transplant setting and that only a proportion of patients will progress to PTLD. Frequent monitoring may help to predict those at highest risk and guide intervention.

Real‐time polymerase chain reaction quantification of Epstein–Barr virus in chronic periodontitis patients

Although herpes viruses have been implicated in the pathogenesis of chronic and aggressive periodontitis, few data in the literature refer to quantification of these viruses in periodontal sites, especially in relation to serological findings. The aim of the present study was to compare Epstein--Barr virus (EBV) DNA load in subgingival specimens from chronic periodontitis patients and in periodontally healthy subjects, in relation to serologic testing of IgM and IgG antibodies to EBV. A total of 22 chronic periodontitis patients and 13 controls participated in the present study. Seventy-nine subgingival specimens (one pooled, one from a deep and one from a shallow site), sampled with paper points, were analysed with real-time polymerase chain reaction for EBV. Subjects were also examined for anti-EBV IgG and IgM levels in serum, using an enzyme-linked immunosorbent assay. One subject was seronegative for EBV. Three subjects (one patient and two controls) displayed anti-EBV IgM. Their data were excluded from further analysis. All three displayed EBV in their subgingival samples. Nine out of the remaining 20 chronic periodontitis patients and 10 out of 11 controls did not display EBV subgingivally. A statistically significant difference in viral load was observed between pooled and shallow-pocket samples from periodontitis patients but not between samples from deep and shallow pockets (Kruskall--Wallis anova, Dunn's multiple comparisons test). Data from the present study do not strongly support the pathogenetic significance of EBV in chronic periodontitis lesions. The data do, however, suggest that parallel serological assessments provide a useful insight into the association of viruses with periodontal disease.

An Unusual Case of Peripheral T-Cell Lymphoma With CD56 Positivity and Angiocentric, Angiodestructive Morphology Arising in the Ileum

Natural killer cell and cytotoxic T-cell lymphomas are frequently difficult to distinguish because they share many common features, and yet it is important to make an accurate diagnosis because their prognoses differ. We report an unusual case of a white man with a CD56-positive T-cell lymphoma in the ileum. The histologic pattern was characterized by angioinvasion and angiodestruction. Immunohistochemical staining showed positive reactions to CD3, CD8, CD43, CD45RO, CD56, and T-cell intracellular antigen-1, but negative reactions to CD4, CD5, CD20, CD23, and CD57. Epstein-Barr virus (EBV) was not detected by EBV-latent membrane protein staining and EBV polymerase chain reaction technique. The T-cell receptor gamma chain gene was rearranged. According to the World Health Organization classification, the absence of EBV excludes the diagnosis of extranodal natural killer/T-cell lymphoma, nasal type. However, the association of EBV with this lymphoma in white patients is not clear. Therefore, absence of EBV alone does not necessarily exclude nasal-type natural killer/T-cell lymphoma, particularly because the histologic pattern in this case is highly characteristic of this tumor.

Predictive value of polymerase chain reaction of cerebrospinal fluid for detection of Epstein-Barr virus to establish the diagnosis of HIV-related primary central nervous system lymphoma.

Polymerase chain reaction (PCR) of cerebrospinal fluid (CSF) for detection of Epstein-Barr virus (EBV) DNA has been proposed as a minimally invasive method for establishing the diagnosis of human immunodeficiency virus-related primary central nervous system lymphoma (PCNSL). In a review of the operational characteristics of this test in our clinical practice, the positive predictive value of CSF PCR for EBV for establishing the diagnosis of PCNSL was only 29%. Of 7 patients with CSF PCR positive for EBV, 2 had PCNSL, and 5 received alternative diagnoses (specificity, 79.1%).

The value of prospective monitoring of Epstein-Barr virus DNA in blood samples of pediatric liver transplant recipients.

Post-transplant lymphoproliferative disease (PTLD) is one of the major causes of morbidity and mortality in transplantation patients. A primary Epstein-Barr virus (EBV) infection is a major risk factor for developing PTLD. The aim of this study was to determine circulating EBV DNA after liver transplantation in pediatric patients in relation to primary EBV infection and development of PTLD. EBV serology was performed before transplantation. Every 4 weeks after transplantation a competitive quantitative polymerase chain reaction (PCR) assay for EBV nuclear antigen-1 was performed in 13 patients. Patients were followed for development of a PTLD. Before transplantation four patients were EBV seropositive and nine patients were EBV seronegative. In one of the four patients who were EBV seropositive before transplantation, EBV DNA became detectable after transplantation, with a peak load of 3600 copies/mL. None of these four patients developed a PTLD. Eight of the nine patients who were EBV seronegative before transplantation developed positive EBV DNA samples. EBV DNA was first detected at a mean of 64 days after transplantation (range 38-89). The mean peak EBV DNA load was 79,700 copies/mL (3600-446,000). Two of these patients developed PTLD, but they could not be identified based on prior or concomitant EBV PCR results. In pediatric liver transplantation EBV DNA load is higher in patients with a primary infection than in patients who were EBV seropositive before transplantation. The EBV PCR cannot be used to identify individual patients who develop PTLD. However, elevated EBV DNA load can be used to detect a group of patients at increased risk for PTLD.

Coinfection by Multiple Strains of Epstein-Barr Virus in Infectious Mononucleosis in Immunocompetent Patients

Objective--A molecular study of Epstein-Barr virus (EBV)-induced infectious mononucleosis (IM) was performed. The presence of a 30-bp deletion on the latent membrane protein-1 (LMP-1) oncogene from EBV in Caucasian IM patients was evaluated. Material and Methods--Peripheral blood mononuclear cells were obtained from 27 IM patients and 18 adenoids, 28 tonsils and 16 EBV-related reactive lymphadenitis specimens were used as controls. DNA isolation, EBV polymerase chain reaction (PCR) and LMP-1 oncogene PCR analysis were performed. Results--The 30-bp deletion on LMP-1 was identified in 29.6% of IM patients, but was always seen in conjunction with full-length LMP-1. Although the LMP-1-deleted strain seemed to be more prevalent in IM (29.6%) and EBV-related reactive lymphadenitis (37.5%) than in adenoid (0%) or tonsil specimens (21.4%), these differences were not significant (p>0.05;χ2). Thus, a 30-bp deletion on LMP-1 was present in almost a third of Caucasian IM patients. Conclusion--The finding of coinfections in IM patients confirms that primary infection by more than one EBV strain is possible.

Serial detection of Epstein–Barr virus DNA in sera and peripheral blood leukocyte samples of pediatric renal allograft recipients with persistent mononucleosis‐like symptoms defines patients at risk to develop post‐transplant lymphoproliferative disease

We tested blood samples of 25 pediatric renal transplant recipients for Epstein-Barr virus (EBV) DNA load by quantitative polymerase chain reaction (PCR). Eleven of these transplant recipients showed clinical persistent mononucleosis-like symptoms years after transplantation (Tx). A quantitation of EBV DNA by PCR in peripheral blood lymphocyte (PBL) and serum samples revealed variable EBV DNA titers. The majority of EBV PCR results in samples of the 14 asymptomatic transplant recipients was repeatedly below detection limit. In contrast, patients with mononucleosis-like symptoms showed persistent EBV genome titers over a period of 6 months, ranging from 75 to 18 750 copies/10 000 PBL and from 680 to 335 000 copies/mL serum, respectively. One child suffering from this mononucleosis-like condition developed an EBV-associated Burkitt-like lymphoma 29 months after Tx. Whereas clinical and histological investigations did not indicate a post-transplant lymphoproliferative disorder (PTLD) until tumor detection, EBV titers in PBL and serum had been high for at least 8 months. We propose that pediatric transplant recipients who show both, recurrent mononucleosis-like symptoms and a sustained high EBV genome load, are at increased risk for severe EBV-related post-transplant complications.

Quantitative tissue polymerase chain reaction for Epstein-Barr virus in pediatric solid organ recipients

Background: Infections caused by herpes virus, in particular, Epstein-Barr virus (EBV), remain a major challenge in solid organ transplantation. Little is known about the significance of tissue EBV load. Methods: Twenty-three tissue biopsy specimens (19 kidney, 3 gastrointestinal, and 1 tonsil specimen) and 2 bronchoalveolar lavage specimens from 14 pediatric transplant recipients (10 kidney, 3 liver, 1 combined transplant) were subject to tissue EBV polymerase chain reaction (PCR) semiquantitative analysis and enzyme-linked immunosorbent assay (ELISA) methods. Results of biopsies were correlated with clinical data. Results: Five of 14 patients had clinically diagnosed EBV disease: 2 patients presented with a septic picture with multiorgan failure and pneumonitis; 1 patient had mononucleosis; 1 patient had an increase in serum creatinine level, lymphadenopathy, and chronic fatigue; and 1 patient had EBV nephritis. These 5 patients underwent 12 biopsies at the time of clinically active infection; 8 biopsies had positive results (up to 111 copies/10 μL of extracted DNA). Conversely, 1 of the remaining 13 tissue biopsy specimens from asymptomatic patients had positive results on ELISA, but undetectable viral load, whereas 8 patients had a positive EBV immunoglobulin G titer with historic evidence of EBV replication in the blood. No patient without evidence of EBV had positive EBV tissue PCR results. Conclusion: Increased EBV load was found in more than 50% of patients, pointing to a previously underrecognized importance of EBV detection in tissues from transplant recipients. The presence of EBV in tissue correlated with the presence of viremia, whereas tissue PCR had 100% specificity. EBV load should be included in biopsy evaluation. Am J Kidney Dis 41:212-219. © 2003 by the National Kidney Foundation, Inc.

Use of EBV PCR for the Diagnosis and Monitoring of Post-Transplant Lymphoproliferative Disorder in Adult Solid Organ Transplant Patients

Epstein-Barr virus (EBV) is known to be involved in the majority of patients who develop post-transplant lymphoproliferative disorder after solid organ transplant. We conducted a retrospective study to determine the utility of qualitative and quantitative Epstein-Barr virus polymerase chain reaction (PCR) for the diagnosis and monitoring of post-transplant lymphoproliferative disorder in adult solid organ transplant patients. Peripheral blood leukocytes obtained from 35 adult solid organ transplant patients consecutively referred for evaluation of possible post-transplant lymphoproliferative disorder, were tested by EBV PCR at the time of initial evaluation and at time points thereafter. Eighteen of 35 (51%) patients were ultimately diagnosed with post-transplant lymphoproliferative disorder by tissue biopsy. Fifteen of 18 (83%) patients were found to have EBER-1 positive tumors by in situ hybridization. EBV PCR was positive in 7 of 15 patients, suggesting a sensitivity of 39%. Seventeen patients without post-transplant lymphoproliferative disorder and three with EBER-1 negative post-transplant lymphoproliferative disorder all had negative EBV PCR tests, suggesting a specificity of 100%. We observed that declines in EBV DNA load were associated with response to therapeutic interventions, such as reduction in immunosuppression, rituximab therapy and chemotherapy. We conclude that peripheral blood EBV PCR may have a role in the diagnosis and monitoring of post-transplant lymphoproliferative disorder in adult solid organ transplant patients.

Research agenda for pediatric gastroenterology, hepatology and nutrition: transplantation. Report of the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition for the Children's Digestive Health and Nutrition Foundation.

Research agenda for pediatric gastroenterology, hepatology and nutrition: transplantation. Report of the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition for the Children's Digestive Health and Nutrition Foundation.

Utility of semiquantitative polymerase chain reaction for Epstein-Barr virus to measure virus load in pediatric organ transplant recipients with and without posttransplant lymphoproliferative disease.

We examined the utility of Epstein-Barr virus (EBV) load as a test for the presence of posttransplant lymphoproliferative disease (PTLD). A semiquantitative (SQ) EBV polymerase chain reaction (PCR) on peripheral blood mononuclear cells (PBMC) was used to determine virus load. We compared the values from pediatric patients, both with and without PTLD, with those from healthy pediatric and adult subjects. The virus loads for asymptomatic healthy subjects had a range of 0-1 log10 cells/10(6) PBMCs. Among transplant recipients (n=135), the mean virus load (+/- standard deviation) at the time of diagnosis of PTLD was 3.1+/-1.2 log(10) cells/10(6) PBMCs versus a baseline value of 1.3+/-1.4 log(10) cells/10(6) PBMCs in children without PTLD (P<.0001). A cutoff of > or =3 log10 cells/10(6) peripheral blood leukocytes resulted in the following values for use of virus load as a test for PTLD: sensitivity, 69%; specificity, 76%; positive predictive value, 28%; and negative predictive value, 95%. We conclude that determination of EBV load by use of SQ PCR is more useful in ruling out than in indicating the presence of PTLD.

Epstein-Barr virus detection in ductal carcinoma of the breast.

Epstein-Barr virus detection in ductal carcinoma of the breast.

Fulminant EBV+ T-cell lymphoproliferative disorder following acute/chronic EBV infection: a distinct clinicopathologic syndrome

AbstractThis study describes the clinicopathologic features of 5 patients who developed a fulminant Epstein-Barr virus (EBV)-positive clonal T-cell lymphoproliferative disorder (LPD) after acute EBV infection. One additional patient developed a similar disorder in the setting of long-standing chronic active EBV infection. Detailed immunophenotyping, in situ hybridization for EBV early RNA-1 (EBER1) and polymerase chain reaction (PCR) analyses for immunoglobulin (Ig) heavy chain and T-cell receptor (TCR)-gamma gene rearrangements were performed on paraffin-embedded tissue from all patients. In addition, EBV strain typing and detection of the characteristic 30-bp deletion of the latent membrane protein-1 (LMP-1) gene were performed by PCR. Controls included 8 cases of uncomplicated infectious mononucleosis (IM). Patients included 4 males and 2 females with a median age of 18 years (2-37 years). Three patients were Mexican, 2 were white, and 1 was of Asian descent. All presented with fever, hepatosplenomegaly, and pancytopenia; 5 were previously healthy, but had a clinical history of a recent viral-like upper respiratory illness (1 week to 2 months), and 1 patient had documented chronic active EBV infection for 7 years. Serologic data for EBV were incomplete but titers were either negative or only modestly elevated in 3 cases. In 1 case serology was consistent with severe chronic active EBV infection. In the remaining 2 cases serologic studies were not performed. All patients died within 7 days to 8 months of presentation with T-cell LPD. On histologic examination, the liver and spleen showed prominent sinusoidal and portal lymphoid infiltrates of CD3+, βF1+, EBER1+ T cells lacking significant cytologic atypia. Two cases were CD4+, 2 cases were CD8+, and 2 cases had admixed CD4+ and CD8+ cells without clear subset predominance. All were TIA-1+, CD56−. Only rare B cells were noted. Marked erythrophagocytosis was present. Molecular analysis revealed identical T-cell clones in 2 or more sites (liver, spleen, lymph node) in 5 cases. All patients carried type A EBV; 4 cases had wild-type EBV-LMP, and 2 showed the 30-bp deletion. This fulminant T-cell LPD after acute/chronic EBV infection is characterized by hepatosplenomegaly, often without significant lymphadenopathy, fever, liver failure, pancytopenia, and erythrophagocytosis indicative of a hemophagocytic syndrome. EBV serology may be misleading, with lack of elevated titers. The presence of an EBER1+ T-cell infiltrate with scant B cells should alert one to this diagnosis. Although cytologic atypia is minimal, studies for T-cell clonality confirm the diagnosis.

Fulminant EBV+ T-cell lymphoproliferative disorder following acute/chronic EBV infection: a distinct clinicopathologic syndrome

This study describes the clinicopathologic features of 5 patients who developed a fulminant Epstein-Barr virus (EBV)-positive clonal T-cell lymphoproliferative disorder (LPD) after acute EBV infection. One additional patient developed a similar disorder in the setting of long-standing chronic active EBV infection. Detailed immunophenotyping, in situ hybridization for EBV early RNA-1 (EBER1) and polymerase chain reaction (PCR) analyses for immunoglobulin (Ig) heavy chain and T-cell receptor (TCR)-gamma gene rearrangements were performed on paraffin-embedded tissue from all patients. In addition, EBV strain typing and detection of the characteristic 30-bp deletion of the latent membrane protein-1 (LMP-1) gene were performed by PCR. Controls included 8 cases of uncomplicated infectious mononucleosis (IM). Patients included 4 males and 2 females with a median age of 18 years (2-37 years). Three patients were Mexican, 2 were white, and 1 was of Asian descent. All presented with fever, hepatosplenomegaly, and pancytopenia; 5 were previously healthy, but had a clinical history of a recent viral-like upper respiratory illness (1 week to 2 months), and 1 patient had documented chronic active EBV infection for 7 years. Serologic data for EBV were incomplete but titers were either negative or only modestly elevated in 3 cases. In 1 case serology was consistent with severe chronic active EBV infection. In the remaining 2 cases serologic studies were not performed. All patients died within 7 days to 8 months of presentation with T-cell LPD. On histologic examination, the liver and spleen showed prominent sinusoidal and portal lymphoid infiltrates of CD3+, βF1+, EBER1+ T cells lacking significant cytologic atypia. Two cases were CD4+, 2 cases were CD8+, and 2 cases had admixed CD4+ and CD8+ cells without clear subset predominance. All were TIA-1+, CD56−. Only rare B cells were noted. Marked erythrophagocytosis was present. Molecular analysis revealed identical T-cell clones in 2 or more sites (liver, spleen, lymph node) in 5 cases. All patients carried type A EBV; 4 cases had wild-type EBV-LMP, and 2 showed the 30-bp deletion. This fulminant T-cell LPD after acute/chronic EBV infection is characterized by hepatosplenomegaly, often without significant lymphadenopathy, fever, liver failure, pancytopenia, and erythrophagocytosis indicative of a hemophagocytic syndrome. EBV serology may be misleading, with lack of elevated titers. The presence of an EBER1+ T-cell infiltrate with scant B cells should alert one to this diagnosis. Although cytologic atypia is minimal, studies for T-cell clonality confirm the diagnosis.

Semiquantitative Epstein-Barr virus polymerase chain reaction analysis of peripheral blood from organ transplant patients and risk for the development of lymphoproliferative disease.

To the Editor: Epstein-Barr virus (EBV) is a human herpesvirus associated with B-cell lymphoproliferations occurring after solid organ and stem cell transplantation.[1-4][1] In organ transplant recipients, the use of chronic immunosuppressive medications impairs T-cell surveillance of EBV-induced B

Polymerase chain reaction amplification of archival material for epstein-barr virus, cytomegalovirus, human herpesvirus 6, and parvovirus B19 in children with bone marrow hemophagocytosis

Bone marrow hemophagocytosis may occur as an incidental finding, or it may be a manifestation of a systemic and potentially lethal disorder. When systemic, the proliferation is termed hemophagocytic lymphohistiocytosis (HLH), a clinicopathologic entity characterized by a widespread proliferation of benign hemophagocytic histiocytes, fever, pancytopenia, deranged liver function, and frequently coagulopathy and hepatosplenomegaly. A variety of infectious agents, including Epstein-Barr virus (EBV), cytomegalovirus (CMV), human herpesvirus 6 (HHV6), and parvovirus B19 (PVB19), have been associated with HLH, but the relative frequency of each using one technique has not been evaluated. In addition, infectious causes of incidental bone marrow hemophagocytosis, not occurring in the setting of HLH, have not been evaluated. Review of bone marrow reports from bone marrow examinations done between December 1986 and June 1997 showed that 20 children aged 2 months to 15 years had bone marrow examinations that indicated hemophagocytosis. Archival materials from 19 patients were successfully retrieved, and DNA was extracted from archived unstained coverslips with subsequent polymerase chain reaction for EBV, CMV, HHV6, and PVB19 genomic DNA. DNA extracted from 16 bone marrow specimens of age-matched children was used as negative controls. Eleven of the 19 patients fulfilled the clinical and pathological criteria for HLH; the remaining eight patients had isolated hemophagocytosis without a systemic presentation. Viral DNA was detected in 8 of 11 patients with HLH but in none of eight patients with isolated hemophagocytosis. EBV was present in five of the bone marrows, followed in frequency by HHV6, CMV, and PVB19. Infection with more than one agent was present in three patients. Only one control patient was positive for HHV6 DNA; the remaining control patients were negative for all viruses. Viral infection, detected by PCR analysis of bone marrow, is a common finding in patients with HLH but not in patients with isolated bone marrow hemophagocytosis. This technique may provide another marker to aid in the diagnosis of HLH and suggests a different cause of hemophagocytosis occurring in patients with and without HLH.

Multiple myeloma in an HIV-positive man presenting with primary cutaneous plasmacytomas and spinal cord compression

heavy chains and free kappa light chains. The CD4 cell count was 20/mm3. A biopsy specimen of the abdominal cutaneous nodules showed a tumoral proliferation in the dermis, consisting of round large cells with anisocaryosis of the nucleus containing numerous nucleoli. Immunohistochemical staining of the atypical plasma cells was positive for anti-IgA and κ sera (Fig 2). Epstein-Barr virus (EBV) DNA was detected by polymerase chain reaction (PCR) in the cutaneous biopsy specimen (the negative control for EBV PCR was performed on an EBV-negative B-lymphocyte cell line DG75). Immunofluorescence was negative for latency (EBNA2, LMP) and replication proteins (VCA, LMA), with a positive control performed on an EBV-positive B-lymphocyte cell line (B95). In situ hybridization was negative for EBER small RNA detection. A diagnosis of multiple myeloma was considered. Histologic examination and immunohistochemical staining of the skin biopsy specimen led to a diagnosis of secondary EMP secreting IgA κ. The skin involvement progressed rapidly, and a rapidly growing third nodule appeared in the left temporal region. Chemotherapy with melphalan and cortisone was initiated. After 3 cycles, the initial plasmacytomas had grown, and numerous other lesions had appeared on his arms, face, and chest. Osteolytic lesions appeared without hypercalcemia or renal failure. The patient died 3 weeks later. There is a broad spectrum of hematologic and immunologic abnormalities in HIV-1 infection. Although the incidence of B-cell–associated lymphomas is increased in HIV infection, fewer than 30 cases of multiple myeloma (MM) have been reported in persons infected with HIV.1-11 Unlike classic plasma cell tumors, the disease often has extramedullary involvement with unusual clinical presentation and a short latency period. Skin involvement is present in only 4% to 8% of HIVnegative patients11 with extramedullary plasmacytomas (EMPs). We describe a case of MM presenting with primary cutaneous plasmacytomas and spinal cord compression in a 53-year-old man positive for HIV.

Semiquantitative Epstein-Barr Virus (EBV) Polymerase Chain Reaction for the Determination of Patients at Risk for EBV-Induced Lymphoproliferative Disease After Stem Cell Transplantation

AbstractEpstein-Barr virus–induced lymphoproliferative disease (EBV-LPD) is a serious and potentially fatal complication after allogeneic stem cell transplantation (SCT). To evaluate levels of EBV DNA in SCT patients, a semiquantitative polymerase chain reaction (PCR) assay was developed. DNA was extracted from peripheral blood leukocytes and diluted, and PCR was performed by using a primer set specific for a well-conserved sequence of the internal repeat 1 region of the EBV genome. Forty-one SCT patients were screened with this method. Thirty-seven patients received allogeneic transplants, of which 18 were T-cell–depleted marrow. Four additional patients received autologous SCT, one of which was T-cell depleted. The mean time of follow-up by EBV PCR was 147 days (range, 47 to 328 days) posttransplant. The range of EBV copies/μg DNA from normal EBV sero-positive donors was 40 to 4,000. Seven patients had ≥40,000 copies of EBV DNA/μg DNA, all of whom were recipients of T-cell–depleted SCT. Five of the seven patients with elevated levels of EBV DNA developed EBV-LPD. Four of these five patients with EBV-LPD had elevated levels of EBV DNA from 1 to 8 weeks before diagnosis. Two patients with EBV-LPD had normal levels of EBV DNA, and two patients with ≥40,000 copies EBV/μg DNA did not develop EBV-LPD. In one patient, clinical resolution of disease correlated with a decrease in EBV DNA and an increase in the level of EBV-specific cytotoxic T-cell precursors. These data indicate that the measurement of EBV viral load with semiquantitative PCR is useful in detecting EBV-LPD in high-risk patients before the onset of clinical symptoms. Because not all patients with elevated levels of EBV DNA develop EBV-LPD, semiquantitative PCR results cannot substitute for clinical, radiographic, and pathological confirmation of this diagnosis.

Semiquantitative Epstein-Barr Virus (EBV) Polymerase Chain Reaction for the Determination of Patients at Risk for EBV-Induced Lymphoproliferative Disease After Stem Cell Transplantation

Epstein-Barr virus–induced lymphoproliferative disease (EBV-LPD) is a serious and potentially fatal complication after allogeneic stem cell transplantation (SCT). To evaluate levels of EBV DNA in SCT patients, a semiquantitative polymerase chain reaction (PCR) assay was developed. DNA was extracted from peripheral blood leukocytes and diluted, and PCR was performed by using a primer set specific for a well-conserved sequence of the internal repeat 1 region of the EBV genome. Forty-one SCT patients were screened with this method. Thirty-seven patients received allogeneic transplants, of which 18 were T-cell–depleted marrow. Four additional patients received autologous SCT, one of which was T-cell depleted. The mean time of follow-up by EBV PCR was 147 days (range, 47 to 328 days) posttransplant. The range of EBV copies/μg DNA from normal EBV sero-positive donors was 40 to 4,000. Seven patients had ≥40,000 copies of EBV DNA/μg DNA, all of whom were recipients of T-cell–depleted SCT. Five of the seven patients with elevated levels of EBV DNA developed EBV-LPD. Four of these five patients with EBV-LPD had elevated levels of EBV DNA from 1 to 8 weeks before diagnosis. Two patients with EBV-LPD had normal levels of EBV DNA, and two patients with ≥40,000 copies EBV/μg DNA did not develop EBV-LPD. In one patient, clinical resolution of disease correlated with a decrease in EBV DNA and an increase in the level of EBV-specific cytotoxic T-cell precursors. These data indicate that the measurement of EBV viral load with semiquantitative PCR is useful in detecting EBV-LPD in high-risk patients before the onset of clinical symptoms. Because not all patients with elevated levels of EBV DNA develop EBV-LPD, semiquantitative PCR results cannot substitute for clinical, radiographic, and pathological confirmation of this diagnosis.