HHV-6 DNA Research Articles

Reduction of Adverse Effects by a Mushroom Product, Active Hexose Correlated Compound (AHCC) in Patients With Advanced Cancer During Chemotherapy—The Significance of the Levels of HHV-6 DNA in Saliva as a Surrogate Biomarker During Chemotherapy

Chemotherapy improves the outcome of cancer treatment, but patients are sometimes forced to discontinue chemotherapy or drop out of a clinical trial due to adverse effects, such as gastrointestinal disturbances and suppression of bone marrow function. The objective of this study was to evaluate the safety and effectiveness of a mushroom product, active hexose correlated compound (AHCC), on chemotherapy-induced adverse effects and quality of life (QOL) in patients with cancer. Twenty-four patients with cancer received their first cycle of chemotherapy without AHCC and then received their second cycle with AHCC. During chemotherapy, we weekly evaluated adverse effects and QOL via a blood test, EORTC QLQ-C30 questionnaire, and DNA levels of herpes virus type 6 (HHV-6) in saliva. The DNA levels of HHV-6 were significantly increased after chemotherapy. Interestingly, administration of AHCC significantly decreased the levels of HHV-6 in saliva during chemotherapy and improved not only QOL scores in the EORTC QLQ-C30 questionnaire but also hematotoxicity and hepatotoxicity. These findings suggest that salivary HHV-6 levels may be a good biomarker of QOL in patients during chemotherapy, and that AHCC may have a beneficial effect on chemotherapy-associated adverse effects and QOL in patients with cancer undergoing chemotherapy.

Understanding the association between chromosomally integrated human herpesvirus 6 and HIV disease: a cross-sectional study

We conducted a cross-sectional investigation to identify evidence of a potential modifying effect of chromosomally integrated human herpes virus 6 (ciHHV-6) on human immunodeficiency virus (HIV) disease progression and/or severity. ciHHV-6 was identified by detecting HHV-6 DNA in hair follicle specimens of 439 subjects. There was no statistically significant relationship between the presence of ciHHV-6 and HIV disease progression to acquired immunodeficiency syndrome. However, after adjusting for use of antiretroviral therapy, all subjects with ciHHV-6 had low severity HIV disease; these findings were not statistically significant. A multi-center study with a larger sample size will be needed to more precisely determine if there is an association between ciHHV-6 and low HIV disease severity.

Prevalence of past and reactivated viral infections and efficacy of cyclosporine A as monotherapy or in combination in patients with psoriatic arthritis--synergy study: a longitudinal observational study.

We have prospectively evaluated psoriatic arthritis (PsA) patients for (1) seropositivity for former viral infections and seroconversion and (2) efficacy of cyclosporine A (CsA) alone or in combination with other immunosuppressants in a time period of 12 months. Screening included HBV antibodies and antigens, HCV antibodies and RNA, HSV 1-2, HZV, EBV, and CMV IgG, and IgM, HHV-6 DNA, and HIV 1-2 antibodies. PsA was evaluated by the Psoriasis Area Severity Index (PASI), the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), and the Visual Analogue Scale (VAS). At baseline, 126 (56%) out of 225 evaluable patients had 2 or more seropositivities indicative of former infections, and 31 patients (13.8%) presented seropositivity for HCV, HBV, HSV-1 and -2, HHV-6, EBV, or parvovirus infection; one of them, positive for HBAg, was treated with lamivudine, while the remaining 30 received no specific treatment. None of the 31 patients developed virus reactivation. A reduction (P < 0.001) of PASI, BASDAI, and VAS scores was observed at 6 and 12 months. The treatment of PsA with CsA as monotherapy or in combination was safe and effective. In vitro experiments and clinical findings, including those from our study, suggest that CsA as monotherapy or in combination with biologics might be the treatment of choice in PsA HCV-positive patients.

Correlation of laryngeal squamous cell carcinoma and infections with either HHV-8 or HPV-16/18

Each year more than 159,000 new cases of laryngeal cancer are diagnosed globally, and more than 9000 patients die due to this malignancy. Viral infections are a known risk factor for this malignancy. Thus, this study aimed to evaluate the role of HPV-16/18 and HHV-8 infection in patients with laryngeal cancer.In this case–control study, 60 formalin-fixed, paraffin-embedded samples of laryngeal cancer and 22 normal larynx tissue samples from the Pathology Department of Qaem Hospital, Mashhad, Iran were studied. After validating the diagnosis, the samples were evaluated for the detection of HPV-16/18 and HHV-8 DNA using PCR technique. The data were registered and analyzed using SPSS 18.0.The average age for patients and controls was 61.29±11.89 and 55.77±10.10, respectively. Fifty-four patients (90%) and 16 (72.7%) controls were male. PCR results detected no HPV-16/18 DNA in both groups. Although there were 2 positive HHV-8 samples in both laryngeal cancer and normal larynx samples, no significant relation was present (p=0.291).We found no significant relationship between infection with HHV-8 or HPV-16/18 and the existence of laryngeal cancer. However, more complementary studies are required to re-evaluate our results using more samples and better viral detection techniques.

Understanding the association between chromosomally integrated human herpesvirus 6 and HIV disease: a cross-sectional study

We conducted a cross-sectional investigation to identify evidence of a potential modifying effect of chromosomally integrated human herpes virus 6 (ciHHV-6) on human immunodeficiency virus (HIV) disease progression and/or severity. ciHHV-6 was identified by detecting HHV-6 DNA in hair follicle specimens of 439 subjects. There was no statistically significant relationship between the presence of ciHHV-6 and HIV disease progression to acquired immunodeficiency syndrome. However, after adjusting for use of antiretroviral therapy, all subjects with ciHHV-6 had low severity HIV disease; these findings were not statistically significant. A multi-center study with a larger sample size will be needed to more precisely determine if there is an association between ciHHV-6 and low HIV disease severity.

Quantification of two viral transcripts by real time PCR to investigate human herpesvirus type 6 active infection

BackgroundHuman herpesvirus 6 (HHV-6) causes exanthema subitum and is associated with symptomatic reactivations in immunocompromised patients, particularly after hematopoietic stem cell transplantation. The detection of viral mRNA can help to make the difference between latent, chromosomally integrated and true replicating virus. It can also be a useful tool to investigate viral multiplication in different cell types. ObjectivesTo develop molecular tools for the detection and quantification HHV-6 transcripts that can be used in a clinical setting. Study-designTwo one-step reverse-transcriptase quantitative real-time PCR (RT-qPCR) were developed for the quantification of the immediate early U90 and the late U100 mRNAs. Viral mRNA loads were compared to viral DNA loads during infection in vitro and in blood samples collected from stem cell transplanted patients. ResultsAnalytical performances of the two quantitative real-time PCR were good. In vitro, kinetics of both transcripts was well correlated with DNA levels. Sixty blood samples from patients with active HHV-6 infection were analyzed. Overall agreement of qualitative results for HHV-6 DNA, U90 RNA and U100 RNA was good. HHV-6 DNA loads were significantly higher than mRNA loads. In clinical samples, the amounts of U100 and U90 mRNAs were low and their detection was mainly associated to viral DNA loads upper than 1000copies/ml of blood. ConclusionThe new assays are sensitive and reliable methods for the monitoring of viral transcription in vitro and in vivo. As their detection is associated to high DNA loads in vivo, they can be helpful tools for the diagnosis of active infection.

Human Herpes Virus 6 Reactivation and Disease After Allogeneic Hematopoietic Stem Cell Transplantation

Background Human herpesvirus type 6 (HHV-6) is a member of the beta herpesvirus subfamily (genus Roseolovirus) and two distinct variants have been described: types A and B. HHV-6 infection is recognized as the cause of a febrile disease and exanthem subitum in early childhood. The infection rarely causes serious events in healthy individuals, but viral reactivation in immunocompromised patients is frequently associated with severe clinical manifestations. Above all HHV-6 is increasingly recognized as an opportunistic and potentially life-threatening pathogen in recipients of allogeneic hematopoietic stem cell transplantation (AlloSCT). Approximately 60% of solid organ transplant and 40% of patients undergoing alloSCT experience HHV-6 reactivation, mainly of variant B. Reported clinical manifestations of HHV-6 infection in transplanted patients are skin rash, interstitial pneumonia, bone marrow suppression and encephalitis. Moreover, some clinical reports suggest that HHV-6 can facilitate the occurrence of severe clinical complications of alloSCT, increasing transplant-related mortality. Methods From January 2009 to February 2013, we retrospectively evaluated 54 consecutive adult patients (median age 50 years) who developed positivity to HHV-6 after alloSCT for high-risk hematological malignancies. Stem cell donors were family haploidentical (37), HLA identical sibling (8), unrelated volunteer (6), cord blood (3). The viral load was determined by quantitative PCR (Nanogen Advanced Diagnostic S.r.L) in cell-free body fluids such as plasma, bronchoalveolar lavage (BAL), cerebrospinal fluid (CSF), bone marrow (BM) aspirates or in gastrointestinal biopsies. Results Median time from alloSCT to HHV-6 reactivation was 34 days (range: 0-705). Thirty-one patients presented HHV-6 positive in plasma, 9/54 in BM, 33/54 in gut biopsies or BAL, 7/54 in CSF. At the time of viral positivity all pts were receiving acyclovir as viral prophylaxis except five. Twenty-nine patients had acute graft versus host disease (GvHD). Twenty-two out of these twenty-nine patients experienced a grade III-IV acute GvHD, requiring high dose steroids in twenty-six cases. A concomitant CMV positivity was detected in 15/54 patients. The median absolute count of CD3+ lymphocytes was 262 cells/mcl. In 52/54 cases we reported HHV-6 clinical manifestations: fever (43), skin rash (22), hepatitis (19), diarrhoea (24), encephalitis (10), BM suppression (18), delayed engraftment (11). HHV-6 positivity led to antiviral pharmacological treatment in 37/54 cases, using as first choice therapy foscarnet. Amongst the total fifty-four patients with documented HHV-6 positivity thirty-one solved the clinical event. However the mortality rate was relatively high in this population (only 30% of patients were alive), mainly related to severe infections or GvHD. A better overall survival is significantly associated with CD3+ cells higher than 200/mcl (p-value 0.011) and time after alloSCT more than 2 months (p-value 0.035). In this analysis the overall survival was not significantly influenced by steroids administration, presence of acute GvHD, plasma viral load and organ involvement. Conclusions This retrospective study further demonstrates the correlation between HHV-6 reactivation and high morbidity and mortality rates in patients after alloSCT. Despite HHV-6 detection typically occurred in the first month after AlloSCT, a better immune reconstitution has the potential to improve the outcome. The regular monitoring of HHV-6 DNA, using a real-time PCR assay, may be useful for identifying active HHV-6 infection and for the introduction of a pre-emptive treatment, possibly reducing the incidence of the most severe clinical complications. Disclosures: Bonini: MolMed SpA: Consultancy.

Analysis of the shedding of three β‐herpesviruses in urine and saliva of children with renal disease

Cytomegalovirus (CMV), human herpesvirus 6 (HHV-6) and 7 (HHV-7) are important pathogens in immunocompromised patients. To elucidate the kinetics of the three β-herpesviruses in saliva and urine samples were collected serially from children with renal diseases. Twenty children with renal diseases were enrolled in this study. A total of 240 saliva and urine samples were collected monthly from the patients over a 1-year period. Viral DNAs loads were measured by real-time PCR. In 10 CMV seropositive patients CMV DNA was detected rarely in saliva and CMV DNA load was lower than the other two β-herpesviruses DNA loads. All patients were seropositive for HHV-6B and the virus was detected frequently in saliva. Two of 20 patients were HHV-7 seronegative. High copies of viral DNA were detected continuously in saliva of the HHV-7 seropositive patients. Although neither CMV nor HHV-6B DNA load was different among the three renal diseases, HHV-7 DNA load was different among the diseases (P = 0.039). HHV-6B DNA loads were significantly higher in patients with immunosuppressive treatment compared to those without treatment (P = 0.013). Although CMV DNA was detected in urine samples collected from 5 of 10 CMV seropositive patients, HHV-6B and HHV-7 DNA were detected at relatively low frequencies in urine. No remarkable temporal associations between viral DNA excretion and proteinuria or immunosuppressive treatment were demonstrated. The pattern of viral DNA excretion in saliva and urine were different among the three viruses. No temporal correlation was observed between viral infection and renal diseases.

HHV-6 antigen and HHV-6 DNA expression in sporadic adenomatous polyps of the colon

Background.Human herpesvirus-6B (HHV-6B) antigens are commonly found in the intestinal mucosa of patients with immunosuppression. In a series of immunocompetent patients with adenomatous, polyp HHV-6B antigen expression from mucosal biopsies was more intense than in biopsies taken from patients receiving immunosuppressive drugs because of kidney transplantation or inflammatory bowel disease. Methods.HHV-6B and cytomegalovirus (CMV) antigen expression was determined from mucosal biopsy samples by immunohistochemistry. HHV-6-DNA content was studied in adenomatous polyps (seven tubular adenomas and one tubulovillous adenoma) taken from eight immunocompetent patients and in three mucosal biopsy samples taken from immunocompetent patients without adenomas using in situ hybridization (ISH) method. Results. HHV-6B antigen expression on mucosal biopsies was strongly positive in five of eight patients with adenomas and negative in all patients without adenoma. CMV antigen expression on mucosal biopsies was faintly positive in three of adenoma patients. HHV-6 ISH was positive in seven of eight adenomatous polyps, most intense in the tubulovillous adenoma and negative in all three mucosal biopsies of patients without adenomas. Conclusion.Intensive HHV-6-DNA expression was found in adenomatous polyps of the colon. Further studies on involvement of HHV-6 in the development of gastrointestinal polyps are warranted.

Human Herpes Virus-6 and Human Herpes Virus -7 in Pityriasis Rosea

**Mersin University School of Medicine, Department of Biostatistics, Mersin, Turkey Objective: The etiology of pityriasis rosea (PR) remains unknown despite numerous investigations. In recent years, human herpes virus-6 (HHV-6) and HHV-7 were accused as causative agents in PR. The aim of this study was to evaluate the possible role of HHV6 and HHV-7 in the pathogenesis of PR. Methods: Twenty three PR patients and 23 healthy blood donors as a control group were included in the study. Polymerase chain reaction (PCR) with specific primers for HHV-6 and HHV-7 DNA sequences was performed on the serum samples of 23 active PR patients and controls, and also the lesional skin biopsies from 11 PR patients. Additionally, serum levels of IgM antibodies againts HHV-7 were detected by using indirect immunofluorescence test on the serum samples of all of the study population. Results: No statistically significant differences were detected between PR patients and controls regarding all serum results. Conclusion: These findings do not support a primary etiological role for HHV-6 and HHV-7 in PR as in some previous studies.

Herpesvirus detection and cytokine levels (IL‐10, IL‐6, and IFN‐γ) in ocular fluid from Tunisian immunocompetent patients with uveitis

Human herpesviruses (HHVs) are involved in the pathogenesis of different types of uveitis. Cytokine response plays an important role in virus-induced immunopathology. This study aimed to investigate the incidence of HHVs in aqueous humor samples of immunocompetent patients with suspected viral uveitis and cytokine (IL-10, IL-6, and IFN-γ) expression profiling. Forty-seven aqueous humor samples were collected from immunocompetent patients with viral uveitis. Samples were assayed for HHV-1 to HHV-8 by in-house real-time polymerase chain reactions. IL-6, IL-10, and IFN-γ were quantified with a cytometric bead array. Relations between viral detection, cytokine profiles, and clinical data were studied. At least one viral genome was detected in 21 aqueous humor samples analyzed. Varicella-zoster virus (VZV) was detected in 14 of the positive samples, cytomegalovirus (CMV) in 8, HSV-1 in 1, Epstein-Barr virus (EBV) in 4, and HHV-6 in 2. More than one viral genome was detected in seven aqueous humor samples. Aqueous humor samples positive for HHV-DNA contained significant levels of IL-6, IL-10, and IFN-γ, compared to HHV-DNA negative samples. High levels of IL-6 were detected in patients with CMV-DNA in their aqueous humor samples. Significantly higher levels of IL-10 and IFN-γ were found in positive samples for VZV, EBV, and HHV-6 DNA than in negative aqueous humor ones. VZV was the principal etiologic agent of uveitis in this Tunisian series, with CMV the second most common agent. Knowledge of immunoregulatory interactions and dynamic changes in viral uveitis may be a key to understand the pathogenesis leading to more-effective treatments.

Human herpesvirus-6 DNAemia is a sign of impending primary CMV infection in CMV sero-discordant renal transplantations

BackgroundHuman herpesvirus-6 (HHV-6) frequently reactivates in immunocompromized individuals. Most commonly HHV-6 DNA is detected without organ localized disease. This HHV-6 DNAemia usually occurs in patients who also have CMV reactivations. The question if reactivation of one virus causes reactivation of the other, or whether both viruses reactivate independently, cannot be answered in populations with high seroprevalence for both viruses. Our study is the first in which 35 patients have been included who were CMV seronegative prior to transplantation. ObjectiveWe investigated the occurrence of HHV-6 reactivations in relation to the CMV-serostatus of renal transplantation donor and recipient. Study design9 consecutive patients receiving a renal transplantation were included. All available stored whole blood samples were tested for HHV-6 DNA by quantitative PCR. Details including CMV serostatus of donor and recipient were recorded. ResultsCMV-seropositive recipients have a 68% chance of developing HHV-6 DNAemia if the kidney came from a CMV-seropositive donor, compared to 26% if the kidney came from a CMV-seronegative donor. CMV-seronegative recipients, who are bound to undergo primary CMV infections following transplantation with a renal graft from a CMV-seropositive donor, have 88% chance of developing HHV-6 DNAemia. If they receive a graft from a CMV-seronegative donor the chance of developing HHV-6 DNAemia is 22%. ConclusionReceiving a renal transplant from a CMV-seropositive donor increases the chance of developing HHV-6 DNAemia. HHV-6 DNAemia is a sign of impending primary CMV infections following sero-discordant renal transplantations.

High seroprevalence of human herpesvirus type 8 in patients with end-stage renal disease in Taiwan

BackgroundHuman herpesvirus type 8 (HHV-8) is the aetiologic agent of Kaposi's sarcoma (KS). The incidence of KS in renal transplant patients is much higher than in healthy controls. The risk is even higher among recipients seropositive for HHV-8 before transplantation. Patients with end-stage renal disease (ESRD) are immunocompromised and are candidates for renal transplantation, but HHV-8 seroprevalence in ESRD patients has not been well documented. ObjectivesThis study aimed to evaluate HHV-8 seroprevalence in ESRD patients in a cohort in Taiwan. Study designBlood samples collected from 149 ESRD patients and 149 age- and sex-matched healthy controls were analysed for HHV-8 antibody with immunofluorescence assay (IFA) and enzyme-linked immunosorbent assay (ELISA) and for HHV-8 DNA with polymerase chain reaction. ResultsSeropositivity and titres for HHV-8 antibodies with IFA as well as seropositivity with ELISA were significantly greater in ESRD patients than in healthy controls (P=0.006, 0.001 and 0.003, respectively). Patients with a history of taking herbal medicine had significantly greater ELISA positivity than those without such a history (P=0.004). ELISA positives, particularly patients, had much higher IFA antibody titres than ELISA negatives (P<0.0001). Seropositivity in ESRD patients was not related to lymphopaenia, monocytosis, dialysis duration or a history of transfusion. Two diabetic ESRD patients were positive for HHV-8 DNA. ConclusionsESRD patients had significantly greater HHV-8 seropositivity than healthy controls in Taiwan. This association seems to be related to the geographic location of the cohort and invites further studies for the early association of HHV-8 infection in ESRD patients and risk for KS.

Frequent detection of cytomegalovirus (CMV) DNA in the lower respiratory tract in CMV‐seropositive pediatric patients with underlying chronic bronchopulmonary diseases lacking canonical immunosuppression

Cytomegalovirus (CMV) may be a relevant cause of morbidity in patients displaying various inflammatory diseases. In this study, it was investigated whether CMV DNA is detected in the lower respiratory tract and the systemic compartment in pediatric patients with chronic or recurrent bronchopulmonary diseases. A total of 42 lower respiratory tract specimens and 11 paired plasma samples from 42 patients were analyzed for the presence of CMV DNA by real‐time PCR. The respiratory specimens were also screened for the presence of respiratory viruses and human herpesvirus 6 (HHV‐6) and 7 (HHV‐7) by PCR methods. Quantitative bacterial and fungal cultures were performed. IL‐6 levels in the respiratory specimens were quantified using ELISA. CMV DNA was detected either in the lower respiratory airways, in plasma, or both in 54.5% of CMV‐seropositive patients. The levels of IL‐6 were significantly higher in these patients than in those with no detectable levels of CMV DNA. HHV‐6 and HHV‐7 DNA were detected in three and one patients, respectively. Respiratory viruses were detected in 13 of the 42 patients. Significant growth of one or more bacterial species was observed in 17 patients. No significant association was found between the presence of CMV DNA and the detection of other microorganisms. The data indicated that the presence of CMV DNA in the lower respiratory tract is a frequent finding in children with chronic or recurrent bronchopulmonary diseases. Further, prospective observational studies are needed to assess the impact of this phenomenon, if any, on the clinical course of these patients. J. Med. Virol. 85:888–892, 2013. © 2013 Wiley Periodicals, Inc.

Animal models for human herpesvirus 6 infection

Human herpesvirus (HHV)-6A and HHV-6B are two enveloped DNA viruses of β-herpesvirus family, infecting over 90% of the population and associated with several diseases, including exanthema subitum (for HHV-6B), multiple sclerosis and encephalitis, particularly in immunosuppressed patients. Animal models are highly important to better understand the pathogenesis of viral infections. Naturally developed neutralizing antibodies to HHV-6 or a related virus were found in different species of monkeys, suggesting their susceptibility to HHV-6 infection. Both HHV-6 DNA and infectious virus were detected in experimentally infected Cynomolgus and African green monkeys, although most animals remained clinically asymptomatic. Furthermore, HHV-6A infection was shown to accelerate the progression of AIDS (acquired immunodeficiency syndrome) in macaques and to lead to the development of neurological symptoms in the marmoset model. Humanized SCID (severe combined immunodeficiency) mice efficiently replicated HHV-6 and were also susceptible to coinfection with HHV-6 and HIV-1 (human immunodeficiency virus 1). As CD46 was identified as a receptor for HHV-6, transgenic mice expressing human CD46 may present a potentially interesting model for study certain aspects of HHV-6 infection and neuroinflammation.

Maladie de Kikuchi-Fujimoto : une présentation atypique

IntroductionHistiocytic necrotizing lymphadenitis (Kikuchi-Fujimoto disease) is a rare clinical entity characterized by the association of enlarged lymph nodes in the posterior cervical region and fever. The disease is more frequent in young women. Case reportWe report a 41-year-old African patient who presented with atypical features of Kikuchi's disease including cutaneous lupus, haemophagocytosis, and lymphocytic meningitis. The ethnic origin and the clinical presentation were initially suggestive of tuberculous meningitis. However, microbiological analyses remained negative, histological findings were suggestive of Kikuchi's disease and HHV6 DNA integration was documented in our patient. ConclusionKikuchi's disease should be suspected in an African patient when lymphocytic meningitis is associated with enlarged cervical lymph nodes, hemophagocytosis and HHV6 DNA integration.

Severe hyponatremia caused by syndrome of inappropriate secretion of antidiuretic hormone developed as initial manifestation of human herpesvirus‐6–associated acute limbic encephalitis after unrelated bone marrow transplantation

Severe hyponatremia is a critical electrolyte abnormality in allogeneic stem cell transplantation (allo-SCT) recipients and >50% of cases of severe hyponatremia are caused by the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). Here, we present a patient with rapidly progressive severe hyponatremia as an initial sign and symptom of human herpesvirus-6-associated post-transplantation acute limbic encephalitis (HHV-6 PALE) after allo-SCT. A 45-year-old woman with acute lymphoblastic leukemia received unrelated bone marrow transplantation from a one locus-mismatched donor at the DR locus. On day 21, she developed a generalized seizure and loss of consciousness with severe hyponatremia, elevated serum antidiuretic hormone (ADH), and decreased serum osmolality. A high titer of HHV-6 DNA was detected in cerebrospinal fluid. Treatment with foscarnet sodium and hypertonic saline was started with improvement of neurological condition within several days. Although an elevated serum ADH, low serum osmolality, and high urinary osmolality persisted for 2months, she had no other recurrent symptoms of encephalitis. Our experience suggests that hyponatremia accompanied by SIADH should be recognized as a prodromal or concomitant manifestation of HHV-6 PALE, and close monitoring of serum sodium levels in high-risk patients for HHV-6 PALE is necessary for immediate diagnosis and treatment initiation.

HHV-8 Related Castleman Disease in the Absence of HIV Infection

AbstractAbstract 1578 Background:Castleman Disease (CD) is a rare atypical lymphoproliferative disorder of unknown aetiology that covers an extremely wide clinical spectrum. It ranges from a localized disease with benign outcome after surgical removal to a life threatening multicentric disease (MCD). In the context of HIV infection, CD is almost always multicentric and linked to HHV-8. Clinical, pathological and laboratory features of HIV+ MCD are well described. In contrast, there is no published series of HHV-8 related CD in HIV negative patients. Methods:From January 1995 through June 2012, we identified in a single centre 55 HIV seronegative patients with pathologically proven CD (25 localized and 30 multicentric). Among them 18 were related to HHV-8. We report on their clinical, pathological and laboratory features. Results:All cases were multicentric. Patients were aged 42–83 years old (median 65,9) and were referred with a relapsing remitting syndrome of fever (94%), general status alteration (100%), peripheral lymphadenopathy (100%), splenomegaly (72%), hepatomegaly (50%) and oedema (28%). Kaposi Sarcoma was observed in 9 cases. Anaemia was a constant finding. White blood cells abnormalities were noted on blood smear during the flares in 13/16 patients. Serum markers of inflammation were present in all cases: a high level of C reactive protein (median 111mg/L; range 36–423), hypergammaglobulinemia (median 31g/L; range 7,3–55), hypoalbuminemia (median 32g/L; range 16–52). PCR for HHV-8 DNA was positive on blood samples in all cases (median 5,5 Log10 copies of KSHV DNA/106PBMCs; range 2,3–6,6), whereas only 12/16 patients tested had a positive HHV-8 serology at diagnosis. All cases showed the classic histological features of the mixed cell type or of the plasma cell type, and LANA-1 immunostaining identified HHV-8 infected plasmablasts in 16/16 tested cases. Reactive hemophagocytic syndrome (44%), Autoimmune Haemolytic Anaemia (33%) and lymphoma (22%) (2 Primary Effusion Lymphoma, 1 lymphoplasmacytic lymphoma and 1 unproved) were the commonest associated complications. Ten patients (55%) have received corticosteroids (CS). CS had little or no effect on lymphoid organs enlargement, inflammatory markers or hemophagocytic syndrome. Occurrence or worsening of KS appeared in one patient each soon after corticosteroids introduction. Remission was obtained with low dose and usually single agent chemotherapy with etoposide in 13/16 cases. Nevertheless, etoposide had only suspensive effect in most cases. Thirteen patients were treated with rituximab, all of them achieved prolonged remission off therapy. After a median follow up of 18 months (range 1–99) after rituximab treatment, only 2 patients experienced relapses. These relapses were successfully retreated with the association of rituximab and etoposide. Fatal outcome occurred in 3 patients not treated with rituximab. Altogether, these features were similar to those described in HIV+ HHV-8 related MCD. On the contrary, comparison between these 18 cases and 12 HIV- HHV-8 unrelated MCD cases showed marked discrepancies. Arthritis, cutaneous manifestations, renal disease, lupus symptoms or POEMS symptoms were more frequently observed in patients with HHV-8 negative MCD. Conclusion:HHV-8 associated MCD should be considered as a single clinicopathological entity whatever the HIV status is. This is a step forward towards a better delineation of CD clinical spectrum, and subsequently of HHV-8 related lymphoproliferative disorders. It also offers the opportunity to improve clinical management of such patients, as it appears that the treatments used in HIV+ HHV-8 related MCD are efficient in HIV- HHV-8 related MCD. Disclosures:No relevant conflicts of interest to declare.

Human Herpesvirus-6 and Cytomegalovirus Dna Infection in Liver Donor Biopsies

Background: Herpesvirus reactivation is common after liver transplantation and co-infections with these viruses may manifest as febrile syndromes. A seropositive donor coupled with a seronegative recipient is a risk factor for primary CMV infection. Objective: Our study analyzed the presence of CMV and HHV-6 - DNA in liver donor biopsies. Methods: Seventy-one patients submitted to orthotopic liver transplant, were investigated for the presence of CMV and HHV-6 DNA and coinfection (both viruses' DNA in the same biopsy) in their liver donor biopsies, using the nested- PCR technique. The biopsies were collected from the organ before the transplant. The clinical donors' information was obtained by the State Health Transplant System, Sao Paulo, Brazil. Results: Forty-four (62%) donors were men and 27 (38 %) were women, almost all the donors were blood group O, with mean age from 18-50. Thirty-seven (52.1%) donors died due to traumatic brain injury. Nine (12.8%) biopsies were positive for CMV DNA and 32 (45%) were positive for HHV-6 DNA. The co-infection (CMV and HHV-6) was positive in 7/71 liver donor biopsies. Conclusions: Infection with CMV and HHV-6 as well as co-infection CMV/HHV-6 has been shown to be highly prevalent in liver donors studied representing a possible risk factor for opportunistic infections.

Concurrent and Fatal HHV-8 Positive Multicentric Castlemanʼs Disease and Kaposiʼs Sarcoma in a HIV Negative Kidney Transplant Recipient

Castleman's disease is a rare lymphoid disorder. It comprises two pathological entities. These are the hyaline-vascular type, which is usually localized (uni-centeric), and the plasma cell type which is usually multicenteric and rather aggressive. Castleman's disease has been described thrice after kidney transplantation. A 62-year-old Caucasian male with a history of kidney transplant presented high fever, night sweats, dyspnoea, coughing, weakness, hepatosplenomegaly, peripheral lymphadenopathy and symptoms of renal failure within 6 years of the transplantation. Laboratory findings revealed anaemia (haemoglobin = 4.6 mmol/l), thrombocytopenia (101 × 109/L), normal WBC total and differential count, renal function impairment (creatinine = 218 μmol/l), hypoalbuminemia (24 g/l), and elevated CRP (133 mg/l). Diagnostic assays for HIV-1 were performed with repeatedly negative results. No acute CMV or EBV infection was detected (by serology and PCR analysis). The clinical diagnosis at this stage was post-transplant lymphoproliferative disorder, therefore dosage of cyclosporin immunosuppression was decreased. CT-scans revealed hepatosplenomegaly and enlarged abdominal lymph nodes. A bone marrow biopsy showed an active haematopoiesis in all cell lineages without any abnormalities. Histological examination of an axillary lymph node biopsy showed a multifocal proliferation of spindle cells. The diagnosis of Kaposi's sarcoma was confirmed by positive staining of these spindle cells with antibodies against factor VIII-related antigen (not shown) and HHV-8. The remainder of the lymph node showed partly involuted germinal centres, surrounded by small lymphocytes. A number of these lymphocytes also stained positive for HHV-8, as seen in multicentric Castleman's disease. The clinical syndrome and the histological findings were compatible with KS and MCD caused by HHV-8 infection in a long-term immunocompromised patient. HHV-8 DNA load in serum or plasma was determined by real-time quantitative PCR (15 × 105 copies/ml). His condition deteriorated rapidly and he died despite therapy. The unicentric Castelman's disease, histopathologically described as the hyaline-vascular subtype, is more common, it is not associated with HHV8-infection, and can mostly be treated effectively with radiotherapy or surgery. The multicentric form is not well understood: it is an atypical lymphoproliferative disorder of a plasma cell type, and is related to immune dysfunction. Patients with MCD often develop malignancies like Kaposi's sarcoma and non-Hodgkin's lymphoma. Clinically, HHV8-positive MCD is more aggressive, and has a poorer prognosis with a median survival of less than 30 months. MCD, whether or not HHV8-associated, is generally treated with combination chemotherapy and steroids.