Reactivation Of Herpesvirus Research Articles

Risk Factors for Human Herpesvirus 6 Reactivation and its Relationship With Syndrome of Inappropriate Antidiuretic Hormone Secretion After Stem Cell Transplantation in Pediatric Patients

There have been several reports on the reactivation of human herpesvirus 6 (HHV6) after stem cell transplantation (SCT) in adults, which sometimes induces severe illness. Few reports exist on pediatric patients; therefore, we retrospectively examined HHV6 reactivation after SCT in children. We reviewed 80 patients with a median age of 6 years. We analyzed HHV6 DNA serum samples from the patients before SCT and at 20 and 40 days after SCT using polymerase chain reaction. We also analyzed the relationship between HHV6 reactivation and the syndrome of inappropriate antidiuretic hormone secretion (SIADH). At 20 days after SCT, 35.0% of serum samples were positive for HHV6 DNA. The median viral load was 3.1×10 copies/mL serum. Multivariate analysis showed cord blood transplantation as the only risk factor for HHV6 reactivation. HHV6 reactivation occurs in 59.4% of 32 patients who underwent cord blood transplantation and in 18.8% of 48 patients who underwent SCT from other sources. Among the 14 patients with SIADH, 78.6% experienced HHV6 reactivation. Among the 66 patients without SIADH, only 25.8% had HHV6 reactivation. This result was statistically significant (P<0.001). This analysis revealed that HHV6 reactivation occurs in many children. In addition, HHV6 reactivation plus SIADH should prompt evaluation for central nervous system disease.

Attachment anxiety is related to Epstein-Barr virus latency.

Attachment theory provides a framework for understanding individual differences in chronic interpersonal stress. Attachment anxiety, a type of relationship insecurity characterized by worry about rejection and abandonment, is a chronic interpersonal stressor. Stress impacts cellular immunity, including herpesvirus reactivation. We investigated whether attachment anxiety was related to the expression of a latent herpesvirus, Epstein-Barr virus (EBV), when individuals were being tested for breast or colon cancer and approximately 1 year later. Participants (N=183) completed a standard attachment questionnaire and provided blood to assess EBV viral capsid antigen (VCA) IgG antibody titers. Individuals with more attachment anxiety had higher EBV VCA IgG antibody titers than those with less attachment anxiety. The strength of the association between attachment anxiety and antibody titers was the same at both assessments. This study is the first to show an association between latent herpesvirus reactivation and attachment anxiety. Because elevated herpesvirus antibody titers reflect poorer cellular immune system control over the latent virus, these data suggest that high attachment anxiety is associated with cellular immune dysregulation.

Changes in nucleosome occupancy occur in a chromosome specific manner.

In the eukaryotic nucleus, DNA is packaged into chromatin. The fundamental subunit of chromatin is the nucleosome, DNA is wrapped 1.6 times around a histone octamer core. Nuclear processes in eukaryotes are impacted by whether regulatory DNA is occupied by nucleosomes. We used microarrays to measure nucleosome occupancy in human cells post-Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation at hundreds of immunity-related loci. The detailed analysis of these technologies can be found in recent publications from our lab [1,3]. We found that nucleosome redistributions displayed chromosome specific nucleosome occupancy. This resource can be used to map nucleosome distributions in a variety of biological contexts.

Reactivation of multiple viruses in patients with sepsis.

A current controversy is whether patients with sepsis progress to an immunosuppressed state. We hypothesized that reactivation of latent viruses occurred with prolonged sepsis thereby providing evidence of clinically-relevant immunosuppression and potentially providing a means to serially-monitor patients' immune status. Secondly, if viral loads are markedly elevated, they may contribute to morbidity and mortality. This study determined if reactivation of herpesviruses, polyomaviruses, and the anellovirus TTV occurred in sepsis and correlated with severity. Serial whole blood and plasma samples from 560 critically-ill septic, 161 critically-ill non-septic, and 164 healthy age-matched patients were analyzed by quantitative-polymerase-chain-reaction for cytomegalovirus (CMV), Epstein-Barr (EBV), herpes-simplex (HSV), human herpes virus-6 (HHV-6), and TTV. Polyomaviruses BK and JC were quantitated in urine. Detectable virus was analyzed with respect to secondary fungal and opportunistic bacterial infections, ICU duration, severity of illness, and survival. Patients with protracted sepsis had markedly increased frequency of detectable virus. Cumulative viral DNA detection rates in blood were: CMV (24.2%), EBV (53.2%), HSV (14.1%), HHV-6 (10.4%), and TTV (77.5%). 42.7% of septic patients had presence of two or more viruses. The 50% detection rate for herpesviruses was 5–8 days after sepsis onset. A small subgroup of septic patients had markedly elevated viral loads (>104–106 DNA copies/ml blood) for CMV, EBV, and HSV. Excluding TTV, DNAemia was uncommon in critically-ill non-septic patients and in age-matched healthy controls. Compared to septic patients without DNAemia, septic patients with viremia had increased fungal and opportunistic bacterial infections. Patients with detectable CMV in plasma had higher 90-day mortality compared to CMV-negative patients; p<0.05. Reactivation of latent viruses is common with prolonged sepsis, with frequencies similar to those occurring in transplant patients on immunosuppressive therapy and consistent with development of an immunosuppressive state. Whether reactivated latent viruses contribute to morbidity and mortality in sepsis remains unknown.

EBV &amp; HHV6 reactivation is infrequent and not associated with MS clinical course

Among the environmental factors associated with multiple sclerosis (MS) causation, some of the strongest associations are with Epstein-Barr virus (EBV), and to a lesser extent human herpesvirus 6 (HHV6). Associations with clinical course are less conclusive, however. We evaluated serum anti-EBV-EA-R IgG and anti-HHV6 IgM, and EBV and HHV6 viral load (VL) for their associations with relapse, disability, and progression in disability in a prospective cohort of 198 participants with clinically definite MS. Anti-EBV-EA-R IgG was detected in 81.8% of cases at study entry, and titers remained essentially unchanged during the study. Anti-HHV6 IgM was detected in only one participant, and EBV-VL (29%) and HHV6-VL (1.8%) were detected in a minority of samples, and where detected levels were low. Our previously demonstrated association between anti-HHV6 IgG and relapse hazard was not affected by adjustment for parameters of reactivation. We found no evidence that any of the viral markers were associated with disability or progression in disability. In relation to relapse, only EBV-VL was positively associated, although this was strongly influenced by a single individual. Using a prospective cohort design, we found no convincing evidence that reactivation parameters of EBV or HHV6 were associated with subsequent MS relapse hazard or progression in disability, confirming previous findings, and indicating that herpesvirus reactivation is not an important driver of relapse or disability in this established MS population.

PDB9 - Effect of thyroid hormone levels on alpha human herpes viruses latency and reactivation

PDB9 - Effect of thyroid hormone levels on alpha human herpes viruses latency and reactivation

The dynamics of herpesvirus reactivations during and after severe drug eruptions: their relation to the clinical phenotype and therapeutic outcome.

Drug-induced hypersensitivity syndrome/drug rash with eosinophilia and systemic symptoms (DIHS/DRESS) and Stevens-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) represent contrasting poles of severe drug eruptions, and sequential reactivations of several herpesviruses have exclusively been demonstrated in the former. No previous studies, however, were extended beyond the acute stage. We sought to investigate whether herpesvirus reactivations could also be observed in SJS/TEN and beyond the acute stage of both diseases. Patients with SJS (n = 16), SJS/TEN overlap (n = 2), TEN (n = 10), and DIHS/DRESS (n = 34) were enrolled. We performed a retrospective analysis of Epstein-Barr virus (EBV), human herpesvirus 6 (HHV-6), and cytomegalovirus (CMV) DNA loads sequentially determined by real-time polymerase chain reaction during a 2-year period after onset. Persistently increased EBV loads were detected in SJS during the acute stage and long after resolution, but not in others. In contrast, high HHV-6 loads were exclusively detected in DIHS/DRESS during the acute stage. The dynamics of herpesvirus reactivation varied in DIHS/DRESS according to the use of systemic corticosteroids: While EBV loads were higher in patients not receiving systemic corticosteroids, CMV and HHV-6 loads were higher in those receiving them. Distinct patterns of herpesvirus reactivation according to the pathological phenotype and to the use of systemic corticosteroids were observed during the acute stage and follow-up period, which may contribute, at least in part, to the difference in the clinical manifestations and long-term outcomes. Systemic corticosteroids during the acute stage may improve the outcomes in DIHS/DRESS.

Inhibition of KAP1 Enhances Hypoxia-Induced Kaposi's Sarcoma-Associated Herpesvirus Reactivation through RBP-Jκ

Hypoxia-inducible factor 1α (HIF-1α) has been frequently implicated in many cancers as well as viral pathogenesis. Kaposi's sarcoma-associated herpesvirus (KSHV) is linked to several human malignancies. It can stabilize HIF-1α during latent infection and undergoes lytic replication in response to hypoxic stress. However, the mechanism by which KSHV controls its latent and lytic life cycle through the deregulation of HIF-1α is not fully understood. Our previous studies showed that the hypoxia-sensitive chromatin remodeler KAP1 was targeted by the KSHV-encoded latency-associated nuclear antigen (LANA) to repress expression of the major lytic replication and transcriptional activator (RTA). Here we further report that an RNA interference-based knockdown of KAP1 in KSHV-infected primary effusion lymphoma (PEL) cells disrupted viral episome stability and abrogated sub-G1/G1 arrest of the cell cycle while increasing the efficiency of KSHV lytic reactivation by hypoxia or using the chemical 12-O-tetradecanoylphorbol-13-acetate (TPA) or sodium butyrate (NaB). Moreover, KSHV genome-wide screening revealed that four hypoxia-responsive clusters have a high concurrence of both RBP-Jκ and HIF-1α binding sites (RBS+HRE) within the same gene promoter and are tightly associated with KAP1. Inhibition of KAP1 greatly enhanced the association of RBP-Jκ with the HIF-1α complex for driving RTA expression not only in normoxia but also in hypoxia. These results suggest that both KAP1 and the concurrence of RBS+HRE within the RTA promoter are essential for KSHV latency and hypoxia-induced lytic reactivation. Kaposi's sarcoma-associated herpesvirus (KSHV), a DNA tumor virus, is an etiological agent linked to several human malignancies, including Kaposi's sarcoma (KS) and primary effusion lymphoma (PEL). HIF-1α, a key hypoxia-inducible factor, is frequently elevated in KSHV latently infected tumor cells and contributes to KSHV lytic replication in hypoxia. The molecular mechanisms of how KSHV controls the latent and lytic life cycle through deregulating HIF-1α remain unclear. In this study, we found that inhibition of hypoxia-sensitive chromatin remodeler KAP1 in KSHV-infected PEL cells leads to a loss of viral genome and increases its sensitivity to hypoxic stress, leading to KSHV lytic reactivation. Importantly, we also found that four hypoxia-responsive clusters within the KSHV genome contain a high concurrence of RBP-Jκ (a key cellular regulator involved in Notch signaling) and HIF-1α binding sites. These sites are also tightly associated with KAP1. This discovery implies that KAP1, RBP-Jκ, and HIF-1α play an essential role in KSHV pathogenesis through subtle cross talk which is dependent on the oxygen levels in the infected cells.

G171(P) Vesicular genital lesions in children – is it sexual abuse?

AimThree cases with vesicular lesions in the ano-genital area are presented. In children this presentation always raises concerns of genital herpes and possible sexual abuse. These cases had varied outcomes...

Increased ratio of FoxP3+ regulatory T cells/CD3+ T cells in skin lesions in drug-induced hypersensitivity syndrome/drug rash with eosinophilia and systemic symptoms

Drug-induced hypersensitivity syndrome/drug rash with eosinophilia with systemic symptoms (DIHS/DRESS) is a severe drug eruption accompanied by multiorgan disorders. Several unique aspects of DIHS/DRESS, including herpesvirus reactivation, liver dysfunction and hypogammaglobulinaemia, have similarities to graft-versus-host disease (GVHD). In this study, we focused on the dynamics of regulatory T cells (Tregs) infiltrating into the skin lesions of DIHS/DRESS and GVHD. Skin biopsies were taken from patients with DIHS/DRESS, GVHD, or maculopapular drug eruption. Tregs were detected using immunostaining with anti-FoxP3. The ratio of FoxP3+ T cells to CD3+ T cells was significantly higher in the skin lesions of patients with DIHS/DRESS than in those of patients with GVHD, and was positively correlated with the number of days from disease onset in the acute phase. The dynamics of Tregs in skin lesions are different between DIHS/DRESS and GVHD, despite there being many similarities between these conditions.

Allopurinol-Induced DRESS Syndrome With a Histologic Pattern Consistent With Interstitial Granulomatous Drug Reaction

To the Editor: We read the case reported by Suran and Christopher1 with great interest because we recently treated a similar case of drug rash with eosinophilia and systemic symptoms (DRESS syndrome) with a histologic pattern consistent with an interstitial granulomatous drug reaction (IGDR). A 45-year-old man was admitted to our department with a pruritic rash and facial edema present for 3 days. The medical history included hyperuricemia; the patient was treated with allopurinol for 2 months. The physical examination demonstrated marked generalized erythema with multiple overlying papules and plaques over the entire body and cervical lymphadenopathy (Figs. 1, 2). The patient had a fever for 2 weeks; the temperature was 39.5°C on admission. Laboratory studies showed a leukocytosis 14.9 (4.0–10.0 × 109/L) with an eosinophil count of 0.81 (0–0.5 × 109/L). The alanine aminotransferase 92 (5–40 IU/L), aspartate aminotransferase 74 (0–40 IU/L), and erythrocyte sedimentation rate 75 (0–10 mm/h) were also elevated. The kidney function testing showed significant abnormalities with a raised blood urea nitrogen 43.5 (7.8–22.0 mg/dL) and creatinine 3.5 (0.6–1.2 mg/dL). The kidney biopsy was consistent with an interstitial nephritis. The skin biopsy specimen taken from a papular lesion on the abdomen revealed a dense, diffuse interstitial, perivascular, and perifollicular infiltrate (Fig. 3A) composed of histiocytes, lymphocytes, eosinophils, and multinucleated giant cells (Figs. 3B, C). Piecemeal fragmentation of collagen and elastic fibers were revealed by Masson-trichrome and Verhoeff–van Gieson stains. There were few mucin deposits noted. Focal vacuolar interface dermatitis was also observed (Fig. 3B). The skin lesions subsided and the liver and kidney function returned to normal after withdrawal of the offending medication, hemodialysis, and steroid therapy.FIGURE 1: Facial edema and erythematous papules.FIGURE 2: Generalized, symmetrical erythema with overlying papules, and plaques on the whole body.FIGURE 3: Histologic findings. A, Dense, diffuse interstitial, perivascular, and perifollicular infiltrates (hematoxylin–eosin, ×12.5). B, Interface dermatitis with basal vacuolopathy (hematoxylin–eosin, ×100). C, Granulomatous infiltrate composed of histiocytes, lymphocytes, eosinophils, and multinucleated giant cells (hematoxylin–eosin, ×200).Allopurinol-induced DRESS syndrome is characterized by hematologic abnormalities, especially eosinophilia and atypical lymphocytosis, skin rash, fever, lymph node enlargement, and single or multiple organ involvement, which starts within 8 weeks after the start of treatment with the offending drug.2,3 Our patient was diagnosed with DRESS syndrome based on fever, lymphadenopathy, cutaneous eruption, eosinophila, and interstitial nephritis developing 8 weeks after starting allopurinol. Histologically, a dense lymphocytic infiltrate in the superficial dermis with eosinophils is frequently observed in patients with DRESS syndrome.2 Diffuse interstitial granulomatous infiltrates with DRESS syndrome, as seen in our patient, were reported by Suran and Christopher.1 IGDR is a rare drug associated entity, initially described in 1998.4 This reaction has been reported in response to calcium channel blockers, angiotensin-converting enzyme inhibitors, beta-blockers, lipid-lowering agents, antihistamines, anticonvulsants, antidepressants, diuretics, sennoside, herbal medications, strontium ranelate, ganciclovir, and anakinra.4–11 Most patients present with erythematous to violaceous plaques with a predilection for the skin fold areas. Lesions are usually caused by drug exposure over a long period of time, and resolve soon after discontinuation of the offending drug. Histopathologically, IGDR is characterized by diffuse interstitial infiltration of lymphocytes and histiocytes, with piecemeal fragmentation of collagen and elastic fibers. Interface dermatitis is usually noted, and affected hair follicles and acrosyringia have also been reported.4,10 Given the appearance of multiple erythematous papules and plaques in association with allopurinol treatment, the histology, and the resolution within 4 weeks after cessation of the drug, were consistent with the diagnosis of IGDR. Chen et al7 reported a case of IGDR that shared some of the features of DRESS, including erythroderma, eosinophilia, and abnormal lymphocytes. However, the patient did not have constitutional symptoms or internal organ involvement. Interestingly, our case and the case reported by Suran and Christopher1 were similar, both cases appeared to have DRESS syndrome and IGDR. Although the pathogenic mechanisms of allopurinol-induced DRESS syndrome have not been fully elucidated, immunologic, genetic, and other factors associated with drug and metabolite accumulation have been proposed to be correlated with the pathophysiology.12 In addition, the mechanisms underlying IGDR are unknown; however, type IV hypersensitivity has been postulated to play a role.7 Suran and Christopher1 proposed that herpes virus-stimulated T cells may cross-react to drug-derived hapten–protein conjugates. However, our patient did not show reactivation of herpes viruses. In conclusion, allopurinol is widely used for the treatment of hyperuricemia; some patients develop the allopurinol-induced DRESS syndrome. We report an extremely rare case of allopurinol-induced DRESS syndrome that was confirmed histologically to be present with IGDR.

Human Herpesvirus 6 Infection in Febrile Children: Frequency in an Iranian Referral Hospital

Polymerase chain reaction (PCR) tests for virus in blood and saliva are frequently positive in persons with past infection, re-infection with new strains or latency with or without repeated reactivation of human herpesvirus 6 (HHV6). The aim of this study is to determine the frequency of HHV6 infections in children aged two years or under with an initial diagnosis of fever during an evaluation in the paediatric emergency department of the Children’s Medical Center, an Iranian referral hospital, using PCR methodology. In all children, the clinical characteristics noted at the initial evaluation as well as demographic and laboratory findings were obtained. Among 150 patients (91 male, 59 female) admitted to the paediatric emergency department, HHV6 was found in 49 (33%; 14 female [29%] and 35 male [71%]). Rash was seen in 14/49 (29%) of HHV6-positive cases, while 35 cases without rash had a positive PCR test (71%). Seizures were found in 78/150 (52%) patients. There was no significant association between seizures and positive HHV6 results (43% in patients without seizure; 57% in cases that developed seizure). Although standard PCR on samples including blood cannot discriminate between latent and active HHV6 infection, nearly a third of patients (mainly children less than one year old) had HHV6 infection.

Fever, Rash, and Systemic Symptoms: Understanding the Role of Virus and HLA in Severe Cutaneous Drug Allergy

Drug hypersensitivity syndromes such as abacavir hypersensitivity and the severe cutaneous adverse drug reactions have been associated with significant short- and long-term morbidity and mortality. More recently, these immunologically mediated and previously unpredictable diseases have been shown to be associated with primarily class I but also class II HLA alleles. The case of the association of HLA-B*57:01 and abacavir hypersensitivity has created a translational roadmap for how this knowledge can be used in the clinic to prevent severe reactions. Although many hurdles exist to the widespread translation of such HLA screening approaches, our understanding of how drugs interact with the major histocompatibility complex has contributed to the discovery of new models that have provided considerable insights into the immunopathogenesis of severe cutaneous adverse drug reactions and other T-cell-mediated drug hypersensitivity syndromes. Future translation of this knowledge will facilitate the development of preclinical toxicity screening to significantly improve efficacy and safety of drug development and design.

Immune reconstitution inflammatory syndrome in a patient with adult-onset Still's disease: Graft-versus-host-like skin reaction with possible asymptomatic human herpes virus reactivation during steroid tapering

Immune reconstitution inflammatory syndrome (IRIS) was originally reported in a human immunodeficiency virus (HIV) patient. It is an excessive inflammatory reaction to an opportunistic infection that occurs in HIV patients with profound immunosuppression during the reconstitution of the immune system in the initial months of highly active antiretroviral therapy (HAART). Within 1-2 weeks after the initiation of HAART therapy, the serum HIV RNA level is reduced and CD4+ T cell counts are increased, [...]

NOD2/CARD15 Single Nucleotide Polymorphism 13 (3020insC) is Associated with Risk of Sepsis and Single Nucleotide Polymorphism 8 (2104C>T) with Herpes Viruses Reactivation in Patients after Allogeneic Hematopoietic Stem Cell Transplantation

Three NOD2 polymorphisms (single nucleotide polymorphism [SNP]8 [2104C>T, Arg702Trp], SNP12 [2722G>C, Gly908Arg], and SNP13 [3020insC, Leu1007 fsins C]), identified as disease-associated variants in Crohn's disease, have recently been suggested as gene markers of the outcome of hematopoietic stem cell transplantation (HSCT). In the present multicenter study of 464 donor-recipient pairs, we focused on the effect of NOD2 mutation(s) on the risk of infections and acute graft-versus-host disease (aGVHD). The presence of SNP13 in recipients, donors, or both was more frequently seen in patients having sepsis than in those lacking sepsis (9 of 48 versus 33 of 386, P = .046). The presence of SNP8 (recipient and/or donor positive) was associated with a higher rate of Herpes viruses reactivation (17 of 21 versus 86 of 173, P = .007). In the SNP8-positive group, a trend for a higher rate of bacteremia well controlled by antibiotics was found (9 of 10 versus 47 of 81, P = .106). In contrast, the presence of SNP13 in recipient and/or donor resulted in a poor response to antibiotics (5 of 11 versus 9 of 10, P = .042). A statistically significant association between the presence of NOD2 SNPs and acute grade > II GVHD was found in a subgroup of HSCT patients who received transplants from unrelated donors with a myeloablative conditioning regimen that included antithymocyte globulin (ATG). In this subgroup of patients, donor positivity for any SNPs investigated (7 of 18 versus 17 of 113, P = .036) and, independently, only the presence of SNP8 (4 of 8 versus 20 of 123, P = .055) were associated with severe grade ≥ II aGVHD. In conclusion, SNP8 positivity in donors or recipients makes patients more prone to Herpes viruses reactivation and bacteremia but not to sepsis. Septic complications were associated with SNP13 polymorphism. SNP8 in donors constitutes a risk factor of severe aGVHD, but only if patients received transplants from unrelated donors and received ATG as part of a conditioning regimen.

Herpes reactivation after injection of dermal fillers

Herpes reactivation after injection of dermal fillers

Human herpesvirus-6 and cytomegalovirus DNA in liver donor biopsies and their correlation with HLA matches and acute cellular rejection

Herpesvirus reactivation is common after liver transplantation. ObjectiveAnalyze the presence of cytomegalovirus (HCMV) and human herpesvirus-6 (HHV-6) DNA in liver donor biopsies, seeking to better understand issues involving human donor leukocyte antigens (HLA)-A, B and DR, as well as correlations with acute cellular rejection. MethodsFifty-nine liver transplantation patients were investigated for the presence of HCMV and HHV-6 DNA in liver donor biopsies, using the Nested-PCR technique. The clinical donor information and HLA matches were obtained from the São Paulo State Transplant System. The recipients’ records regarding acute cellular rejection were studied. ResultsSeven (11.8%) biopsies were positive for HCMV DNA and 29 (49%) were positive for HHV-6 DNA. In 14 donors with HLA-DR 15 nine had HHV-6 DNA positive liver biopsy with a tendency for significant association (p=0.09), 22 recipients developed acute cellular rejection and 9/22 were positive for HLA-DR 15 (p=0.03; χ2=4.51), which was statistically significant in univariate analysis and showed a tendency after multivariate analysis (p=0.08). ConclusionHHV-6 DNA was prevalent in liver donors studied as well as HLA-DR 15. These findings suggest that patients with HLA-DR 15 in liver donor biopsies develop more rejection after liver transplantation.

Higher Risk Of HHV6 Reactivation Among Patients Undergoing Umbilical Cord Blood Transplantation

Introduction Human herpesviruses may cause severe complications after allogeneic hematopoietic stem cell transplantation (HSCT) such as interstitial pneumonia, encephalitis, delayed engraftment and post-transplant lymphoproliferative disease. A prospective survey on the incidence of primary infection or reactivation and clinical features of the eight human herpesviruses after HSCT has not yet been performed in Brazilian patients. Additionally, the impact of most of these infections on the transplant outcome is still unclear. Methods Between August 2010 and December 2012, peripheral blood samples from 99 allogeneic HSCT recipients were collected weekly after transplant until day +100, totalizing 824 samples. Median age was 15 years (range: 2-72), 60% were male, and acute leukemias were the most frequent diagnosis (54%). Stem cell sources were bone marrow in 62%, umbilical cord blood in 22% and mobilized peripheral blood in 16%. Fifty–one percent of donors were related. In a semi-automated workflow, the DNA was extracted from plasma in the QIAcube robot. A test based on quantitative real-time PCR (Taqman®) was optimized to screen and quantify all known human herpesviruses (CMV, EBV, HSV1, HSV2, VZV, HHV6, HHV7 and HHV8). The PCR reactions were set up using QIAgility robot for high-precision pipetting, and have been performed in a 7900HT (Life Technologies). Infected cell cultures and plasma specimens with a known viral load/amplicon copy number have been used as controls. The limit of detection of the qPCR was 5 copies per reaction, representing 250 copies/mL of plasma for all of the viruses. Results The incidences of primary infection or reactivation of herpesviruses were as follows: CMV=41%, HHV6=11%, HHV8=5.5%, EBV=3%, HSV1=3%, VZV=3%, HHV7=2%, and HSV2=1%. CMV reactivation was significantly more frequent in adults (72% vs. 27% for children, p&lt;0.0001), and in those or receiving fludarabine (60% vs. 29%, p=0.03) and TBI (68% vs. 32%, p=0.01) in the conditioning regimen, but in a multivariate analysis, only age greater than 18 years remained significant (HR 3.4, 95%CI 1.7-6.7). HHV6 reactivation was significantly more frequent after umbilical cord blood transplant than after transplant from other sources (41% vs. 6%, respectively, p&lt;0.0001) and in those receiving TBI in the conditioning regimen (19% vs. 3%, p=0.01) and in those receiving mycophenolate mofetil as GVHD prophylaxis (22% vs. 2%, p=0.004). In a multivariate analysis, only the use of cord blood remained significantly associated with the risk of HHV6 reactivation (HR 5.7, 95%CI 1.2-26.2). CMV reactivation was associated with a higher risk of acute graft-versus-host disease (GVHD), with a cumulative incidence at 100 days of 37% vs. 17% (p=0.02), but had no impact on the other outcomes. HHV6 reactivation had no significant impact on outcomes. HHV8 reactivation was associated with an increased risk of chronic GVHD (83% vs. 49%, p=0.001). Conclusion HHV6 primary infection or reactivation is more frequent after umbilical cord blood transplantation. CMV and HHV6 primary infection or reactivation are frequent after HSCT, but had no significantly impact on the transplant outcomes, possibly due to monitoring and preemptive measures. Monitoring these viruses constitute an essential measure to improve outcomes. Disclosures: No relevant conflicts of interest to declare.

The Ribvd Regimen (Rituximab IV, Bendamustine IV, Velcade SC, Dexamethasone IV) Offers a High Complete Response Rate In Elderly Patients With Untreated Mantle Cell Lymphoma. Preliminary Results Of The Lysa Trial “Lymphome Du Manteau 2010 SA”

This prospective phase II trial (Nov 2011- Dec 2012), supported by the LYSA group, aimed to evaluate the impact on PFS of the RiBVD regimen in newly diagnosed, previously untreated, elderly MCL patients (>65 years or not eligible for ASCT) (NCT01457144). Inclusion criteria were: WHO 2008 MCL not previously treated, CD20 positive, ECOG 0-2, AA stage II-IV, no CNS involvement or active HBV/HCV/HIV infection. Patients were scheduled to receive a total of 6 cycles of the RiBVD regimen, if they responded (IWG criteria) after 4 cycles. The regimen was administered every 4 weeks with Rituximab 375 mg/m² IV on day(D)1, Bendamustine 90 mg/m² IV on D1 and 2, Dexamethasone 40 mg/m² IV on D2 and bortezomib (Velcade®) 1.3 mg/m² subcutaneously on D1, 4, 8 and 11. Primary prophylaxis with acyclovir was mandatory for Herpes virus reactivation, but there was no recommendation for bacterial prevention. Herein we present preliminary analysis of the trial after 4 RiBVD cycles. Results: A total of 76 patients were included, one was excluded because of HBV active disease and 5 had insufficient data reported in the database. To date we analyzed 70 patients. Patients characteristics: sex ratio M/F 49/21, median age 72 years (y) [64-83] (2 patients were 64 y old), AAstage II/III-IV 5/65, ECOG 0-1/2 59/11, MIPI score low/intermediate/high 3/19/48. Response: 61 responded (ORR=87%), with 19 in PR (26%) and 42 in CR/CRu (60%). Four patients died from pneumonia (n=1), cardiac arrest (n=2) and one following Progressive Multifocal Leukoencephalopathy. Three patients have progressed after 3 cycles. Sixty one patients were analysed by PETscan after 4 cycles, 39 (64%) reached a CR (30 were in CR/CRu and 9 in PR) and 22 remained PET positive (11 patients were in CR/CRu, 10 in PR and 1 stable). RiBVD cycles: 271 cycles were administered out of 280 planned (97%). Twenty four (9%) were delayed, 10 for toxicity. All but one planned Bendamustine doses (n=542) were administered with dosing modified 17 times (3%), mostly for hematological toxicity (n=14). Regarding Bortezomib, 79% (1028/1084) of planned doses were administered, it was prematurely stopped (56, 4%) for neurotoxicity (10 instances) or hematological side effects (46). Rituximab was not administered in 4 instances. Hematologic toxicities: Over the 271 cycles administered, neutropenia was reported in 104 cycles [56 grade 3/4 (g3/4) (21%)], 2.5% with fever; thrombopenia in 181 cycles [41 g3/4 (15%)]; anemia in 210 cycles, [6 g3/4 (2%)]. Non-hematologic toxicities:Reported in >10% of the cycles were : allergic reactions (10.3%, g3/4 <1%), fatigue (40%, g3/4 5%), fever without neutropenia (12%, g3/4 none), weight loss (12%, g3/4 0%), cutaneous rash (12%, g3/4 1.5%), gastrointestinal (40%, g3/4 1.5% ; diarrhea 8%, constipation 17% or emesis 15%), elevated transaminases (14%, g3/4 1%), creatinin (13%, g3/4 none), or glucose (18%, g3/4 1%) ; neuropathy (sensitive or pain) (25%, g3/4 4%). Serious adverse events (SAE) additional to the 4 deaths, were 12 infections including 3 pneumonias (no pneumocystosis), one listeriosis and 2 herpes zoster. Four febrile neutropenia and 3 cutaneous hypersensitivity episodes were also reported. ConclusionDespite 4 toxic deaths (6%), toxicity appears acceptable and manageable. In particular, subcutaneous bortezomib shows markedly decreased neurotoxicity compared to the IV form. This interim analysis shows that four cycles of RiBVD are very effective for untreated elderly MCL patients (ORR 87%) with a high response rate (CR/CRu 60%) which has been shown to be predictive of a long duration of response. An update of these results will be presented at the ASH meeting together with the molecular response rates after 4 cycles. Disclosures:Gressin:Pfizer: Consultancy; Mundipharma: Consultancy. Cartron:Roche: Membership on an entity's Board of Directors or advisory committees.

Histone deacetylase classes I and II regulate Kaposi's sarcoma-associated herpesvirus reactivation.

In primary effusion lymphoma (PEL) cells infected with latent Kaposi's sarcoma-associated herpesvirus (KSHV), the promoter of the viral lytic switch gene, Rta, is organized into bivalent chromatin, similar to cellular developmental switch genes. Histone deacetylase (HDAC) inhibitors (HDACis) reactivate latent KSHV and dramatically remodel the viral genome topology and chromatin architecture. However, reactivation is not uniform across a population of infected cells. We sought to identify an HDACi cocktail that would uniformly reactivate KSHV and reveal the regulatory HDACs. Using HDACis with various specificities, we found that class I HDACis were sufficient to reactivate the virus but differed in potency. Valproic acid (VPA) was the most effective HDACi, inducing lytic cycle gene expression in 75% of cells, while trichostatin A (TSA) induced less widespread lytic gene expression and inhibited VPA-stimulated reactivation. VPA was only slightly superior to TSA in inducing histone acetylation of Rta's promoter, but only VPA induced significant production of infectious virus, suggesting that HDAC regulation after Rta expression has a dramatic effect on reactivation progression. Ectopic HDACs 1, 3, and 6 inhibited TPA-stimulated KSHV reactivation. Surprisingly, ectopic HDACs 1 and 6 stimulated reactivation independently, suggesting that the stoichiometries of HDAC complexes are critical for the switch. Tubacin, a specific inhibitor of the ubiquitin-binding, proautophagic HDAC6, also inhibited VPA-stimulated reactivation. Immunofluorescence indicated that HDAC6 is expressed diffusely throughout latently infected cells, but its expression level and nuclear localization is increased during reactivation. Overall, our data suggest that inhibition of HDAC classes I and IIa and maintenance of HDAC6 (IIb) activity are required for optimal KSHV reactivation.