Serum levels of matrix metalloproteinase-2, -9, and vitamin D in patients with multiple sclerosis with or without herpesvirus-6 seropositivity

ABSTRACT In recent years, extreme attention has been focused on the role of human herpesvirus-6 (HHV-6) in multiple sclerosis (MS) pathogenesis. However, the pathogenesis of MS associated with HHV-6 infection remains unknown. In this study, we measured the serum levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), and vitamin D levels in MS patients with HHV-6 infection and MS patients without HHV-6 infection. Five hundred sixty (including 300 females and 260 males) MS patients along with 560 healthy subjects were analyzed for HHV-6 seropositivity using enzyme-linked immunosorbent assay (ELISA). Subsequently, we measured the serum levels of MMP-2, MMP-9, and vitamin D levels in MS patients with HHV-6 infection and MS patients without HHV-6 infection by ELISA. About 90.7% of MS patients (508/560) were seropositive for HHV-6, while 82.3% (461/560) of healthy subjects were seropositive for this virus (p = 0.001). Moreover, there was a significant increase in the levels of MMP-2, MMP-9, and lower vitamin D in the serum samples of MS patients when compared with healthy subjects. Additionally, we demonstrated that the MMP-9 levels in seropositive MS patients were significantly higher than seronegative MS patients (p = 0.001). Finally, our results demonstrated that the mean of expanded disability status scale (EDSS) in seropositive MS patients was significantly higher in comparison to seronegative MS patients (p < 0.05). In conclusion, we suggest that the HHV-6 infection may play a role in MS pathogenesis.

Development of an enzyme-linked immunosorbent assay for diagnosis of human herpesvirus-7 infection

Abstract INTRODUCTION: Human herpesvirus (HHV)-7 establishes a latent infection during the lifetime of the host and can reactivate after the primary infection, leading to lytic replication in immunosuppressed patients. METHODS: This study aimed to develop an enzyme-linked immunosorbent assay (ELISA) to identify HHV-7 serum antibodies and compare its performance with that of an indirect immunofluorescence assay (IFA). RESULTS: Serum samples (n=102) were tested by IgG-IFA and by ELISA. IFA and ELISA showed IgG-positive results in 77 and 73 samples, respectively. Qualitative concordance of 96% was demonstrated between the two techniques. CONCLUSIONS: ELISA may be useful to diagnose HHV-7 infection.

Human herpesvirus 6A active infection in patients with autoimmune Hashimoto's thyroiditis

ABSTRACT Background: Hypothyroidism due to Hashimoto's thyroiditis (HT) is the commonest autoimmune endocrine illness in which antibodies against thyroid organ result in inflammation. The disease has a complex etiology that involves genetic and environmental influences. Viral infections may be involved in triggering of the disease as their molecular mimicry enhance autoimmune responses. Human herpesvirus-6 (HHV-6) is recognized for its contribution to some autoimmune diseases. Objective: In the current study, the prevalence of HHV-6 active infection in patients with HT and with non-autoimmune thyroid disorders were compared with patients with euthyroidism. In addition, a correlation between presence of HHV-6 infections and HT was investigated. Methods: A total of 151 patients with clinically and laboratory confirmed HT, 59 patients with non-autoimmune thyroid disorders, and 32 patients with normal thyroid function were included in the study. For further confirmation of HT disease, all the precipitants were tested for anti-thyroid peroxidase (TPO), and anti-thyroglobulin (TG) antibodies. For detection of both HHV-6 types A and B, nested PCR and restriction enzyme digestion were used. HHV-6 DNA positive samples were further investigated by DNA sequencing analysis. Results: HHV-6A DNA was found in serum sample of 57 out of 151 patients (38%) with HT, which was significantly more often than in patients with non-autoimmune thyroid disorders (p = 0.001). However, HHV-6 DNA was not detected in serum samples of euthyroid subjects. Conclusions: The results support a possible role for active HHV-6A infection, demonstrated by the presence of HHV-6 DNA in sera, in the development of HT.

Polirradiculoneuritis aguda asociada a infección por Virus Herpes Humano 7 en un paciente adulto inmunocompetente / Acute polyradiculoneuropathy associated with human Herpes Virus 7 in an immunocompetent patient: case report

Human herpes virus 7 (HHV-7) is a cause of encephalitis, meningitis and myeloradiculoneuropathy in adults who are immunocompetent or with immunosuppression. The involvement of the peripheral nervous system is always associated with myelitis. We report a case of acute polyradiculoneuropathy due to HHV-7, without involvement of central nervous system, in an immunocompetent patient. A 35-years-old man complained of lumbar pain radiating to both buttocks. On examination muscle strength and tendon reflexes were normal. He had asymmetric pinprick and light touch saddle hypoesthesia and also in the perineal region, dorsum and lateral aspect of the left foot. Magnetic resonance imaging showed mild thickening and contrast enhancement of cauda equina nerve roots. Polymerase chain reaction performed on cerebrospinal fluid was positive for HVV-7. Other inflammatory, infectious and neoplastic etiologies were ruled out. Lumbar pain and hypoesthesia improved progressively and neurological examination was normal after one month. He did not receive antiviral therapy.

Optimisation of a quantitative polymerase chain reaction-based strategy for the detection and quantification of human herpesvirus 6 DNA in patients undergoing allogeneic haematopoietic stem cell transplantation

Human herpesvirus 6 (HHV-6) may cause severe complications after haematopoietic stem cell transplantation (HSCT). Monitoring this virus and providing precise, rapid and early diagnosis of related clinical diseases, constitute essential measures to improve outcomes. A prospective survey on the incidence and clinical features of HHV-6 infections after HSCT has not yet been conducted in Brazilian patients and the impact of this infection on HSCT outcome remains unclear. A rapid test based on real-time quantitative polymerase chain reaction (qPCR) has been optimised to screen and quantify clinical samples for HHV-6. The detection step was based on reaction with TaqMan® hydrolysis probes. A set of previously described primers and probes have been tested to evaluate efficiency, sensitivity and reproducibility. The target efficiency range was 91.4% with linearity ranging from 10-106 copies/reaction and a limit of detection of five copies/reaction or 250 copies/mL of plasma. The qPCR assay developed in the present study was simple, rapid and sensitive, allowing the detection of a wide range of HHV-6 loads. In conclusion, this test may be useful as a practical tool to help elucidate the clinical relevance of HHV-6 infection and reactivation in different scenarios and to determine the need for surveillance.

Desenvolvimento de métodos de diagnóstico, silenciamento gênico e caracterização molecular do vírus herpes simples tipo 1 e herpesvírus humano tipo 6 em pacientes imunocomprometidos do Rio de Janeiro / Diagnosis, antiviral therapy and genotypic characterization of the Herpes Simplex type 1 virus (HSV-1) and Human herpesvirus type 6 (HHV-6) in immunocompromised patients in Rio de Janeiro

A família Herpesviridae abriga um grande número de vírus que infectam animais e o homem. Nove herpesvirus foram identificados que possuem o homem como hospedeiro primßrio e descritos através da anßlise filogenética foram classificados em trêssubfamília: Alfaherpesvirinae, Betaherpesvirinae e Gamaherpesvirinae. A família Herpesviridae agrupa vírus de genoma DNA e que estão associados a infecções latentes e a manifestações clínicas recorrentes. As infecções por herpesvírus podem se manifestar de forma assintomßtica ou sintomßtica, podendo causar diferentes síndromes clínicas. Estes vírus normalmente são benigno, sendo o aparecimento de lesões nas mucosas a forma mais comum da doença, podendo causar doenças neurológicas severas principalmente em pacientes imunocomprometidos. A incidência e severidade podem ser exacerbadas entre indivíduos imunocomprometidos. A presente tese é composta por três manuscritos, sendo dois trabalhos publicados e um submetido. Estes estudos se propuseram a padronizar métodos de silenciamento gênico, otimizar e comparar métodos para o diagnóstico e realizar a caracterização genotípica do herpes simples tipo1 (HSV-1) e herpesvírus humano tipo 6 (HHV-6). Os estudos foram realizados em paciente imunocomprometidos do estado do Rio de Janeiro...

The Herpesviridae family has a large number of virus widespread in animals andhuman. Nine herpesvirus are identified that have humans as their primary host anddescribed by phylogenetic analysis were classified into three subfamily:Alfaherpesvirinae, Betaherpesvirinae and Gammaherpesvirinae. The familyHerpesviridae is family of DNA virus and that are associated with latent infections andrecurrent clinical manifestations. Herpesvirus infections can manifest themselves inasymptomatic or symptomatic form, may cause different clinical syndromes. Theseviruses are usually benign and the appearance of mucosal lesions are more common, butsevere neurological diseases can occurred especially in immunocompromised patients. The incidence and severity may be exacerbated among immunocompromisedindividuals. This thesis consists of three manuscripts, two published papers andsubmitted. These studies set out to standardize methods of gene silencing, optimize andcompare methods for the diagnosis and the genotypic characterization of herpes simplextype 1 (HSV-1) and human herpesvirus type 6 (HHV-6)...

Human herpesvirus-6 and Epstein-Barr virus infections at different histopathological grades of oral squamous cell carcinomas

The aim of this study was to determine the prevalence and viral load of Epstein-Barr virus [EBV] and Human herpesvirus-6 [HHV-6] in different histopathologic grades of oral squamous cell carcinoma [OSCC]. Forty-five formalin-fixed paraffin-embedded tissue section of OSCC patients were analyzed by quantitative real-time polymerase chain reaction for detection of EBV and HHV-6. The mean age of the patients was 58.6 years, 69% of whom were female, and 31% were male. Overall, the positive rate for EBV and HHV-6 were 16.7% and 27.1%, respectively; and the mean viral load EBV was 27.9 x 10[3] and 38.5 x 10[3] for HHV-6. No correlation was demonstrated between the viral load of EBV DNA [P = 0.35] and HHV-6 [P = 0.38] at the different OSCC histopathologic grades. These findings neither lend support to the hypothesis that EBV and HHV-6 are directly involved in OSCC nor rule out the possibility that these viruses play an indirect role in carcinogenesis in this area

Early HHV-6 replication is associated with morbidity non-related to CMV infection after kidney transplantation

Human herpesvirus type 6-(HHV-6) has been associated with morbidity after liver transplantation. OBJECTIVE: The aim of this study was to determine the HHV-6 seroprevalence among donor-recipient pairs, analyze the incidence of early active infection, its clinical manifestation, interaction with CMV, and the related morbidity in the first year after kidney transplantation. METHODS: 46 donor-recipient pairs had IgG evaluated by ELISA before transplantation: HHV-6(Pambio - USA) and CMV-(Roche - USA). A frozen whole blood sample collected weekly (from the 1st to the 6th week) was retrospectively tested for HHV-6 viral load (VL) determination by real time quantitative PCR (qPCR, Nanogen - Italy). Patients were preemptively surveyed for CMV by pp65 antigenemia (Ag, APAAP, immunohistochemistry, Biotest - Germany) from the 4th to the 12th week after transplantation. Active infection was defined as qPCR-HHV6+ (viral-load/mL-VL) and Ag+ (+cells/100.000 granulocytes), for HHV-6 and CMV, respectively. DCMV was defined as simultaneous positive antigenemia and suggestive signs/symptoms. Concerning +qPCR-HHV6, associated factors, clinical manifestation, interaction with CMV and morbidity were searched. RESULTS: Pre-transplant HHV-6 seroprevalence was significantly higher among kidney recipients compared to their donors (82.6x54.8%; p = 0.005 [3.9 (1.4-10.4)]). Active infection by this virus occurred in 26.1% (12/46), with no association with previous IgG (p = 0.412). Median VL was 125 copies/mL (53-11.264), and the median Ag was 21 +cells (2-740). There was no association between HHV-6 and CMV activation after transplantation (p = 0.441), neither concerning DCMV (p = 0.596). Median highest Ag+ and days of ganciclovir treatment were similar between qPCR-HHV6 + or - (p = 0.206 and p = 0.124, respectively). qPCR-HHV6+ was associated with higher incidence of bacterial (p = 0.009) and fungal (p = 0.001) infections, and higher number (p = 0.001) of hospital admission and longer duration of hospitalization over the first 6 and 12 months post-transplantation (p = 0.033 and p = 0.001). CONCLUSION: Latent HHV-6 infection is more common among recipients than donors before transplantation. Early active infection by this pathogen after transplantation does not increase DCMV incidence or severity during the first 3 months of follow-up. However, early HHV-6 replication is associated with other infections and hospitalizations in the first year.

Meningomielitis asociadas a infección por herpes virus humano 7: Comunicación de dos casos / Meningomyelitis associated with infection by human herpes virus 7: Report of two cases

Human herpesvirus 7 (HHV-7) may cause encephalomyelitis in immune competent adults. We report two patients infected by the virus. A 34-year-old male presenting with paraparesis and a sensitive deficiency located in D6 dermatome. Cerebrospinal fluid had 35 white blood cells per mm³ and 75 mg protein per dl. A PCR-microarray examination was positive for HHV-7. The patient was treated with prednisolone and ganciclovir with full recovery. A 27-year-old male presenting with headache, fever and diarrhea. Cerebrospinal fluid analysis showed 160 cells per mm³ and 75 mg protein per dl. Viral RNA detection was positive for HHV-7. The patient was managed with analgesia and rest and was discharged with the diagnosis of viral meningitis. Our communication supports the notion that HHV-7 may be considered as pathogen factor in humans, even in immune competent ones.

Simultaneous monitoring of CMV and human herpesvirus 6 infections and diseases in liver transplant patients: one-year follow-up

OBJECTIVE: The aim of this study was to simultaneously monitoring cytomegalovirus and human herpesvirus 6 active infections using nested-polymerase chain reaction and, together with clinical findings, follow the clinical status of patients undergoing liver transplant. INTRODUCTION: The human β-herpesviruses, including cytomegalovirus and human herpesvirus 6, are ubiquitous among human populations. Active infections of human herpesvirus 6 and cytomegalovirus are common after liver transplantation, possibly induced and facilitated by allograft rejection and immunosuppressive therapy. Both viruses affect the success of the transplant procedure. METHODS: Thirty patients submitted to liver transplant at the Liver Transplant Unit, at the Gastro Center, State University of Campinas, SP, Brazil, were studied prospectively from six months to one year, nested-polymerase chain reaction for cytomegalovirus and human herpesvirus 6 DNA detections. Two or more consecutive positive nested-polymerase chain reaction were considered indicative of active infection. RESULTS: Active infection by cytomegalovirus was detected in 13/30 (43.3 percent) patients, median time to first cytomegalovirus detection was 29 days after transplantation (range: 0-99 days). Active infection by human herpesvirus 6 was detected in 12/30 (40 percent) patients, median time to first human herpesvirus 6 detection was 23.5 days after transplantation (range: 0-273 days). The time-related appearance of each virus was not statistically different (p = 0.49). Rejection of the transplanted liver was observed in 16.7 percent (5/30) of the patients. The present analysis showed that human herpesvirus 6 and/or cytomegalovirus active infections were frequent in liver transplant recipients at our center. CONCLUSIONS: Few patients remain free of betaherpesviruses after liver transplantation. Most patients presenting active infection with more than one virus were infected sequentially and not concurrently. Nested-polymerase chain reaction can be considered of limited value for clinically monitoring cytomegalovirus and human herpesvirus 6.

Detection of human herpesvirus 6 and 7 DNA in saliva from healthy adults from Rio de Janeiro, Brazil

In this study, we aimed to evaluate virus shedding in the saliva of healthy adults from the metropolitan region of the city of Rio de Janeiro, Brazil, in order to verify the prevalence of both human herpesviruses 6 and 7 (HHV-6, HHV-7). The studied group comprised 182 healthy individuals at Pedro Ernesto University Hospital, who were being seen for annual odontologic revisions. Saliva specimens were subjected to a multiplex polymerase chain reaction (PCR) to detect the presence of HHV-6A, HHV-6B and HHV-7. The total Roseolovirus DNA prevalence was 22.4 percent. The PCR detected a HHV-6 prevalence of 9.8 percent, with HHV-6A detected in 7.1 percent of the samples and HHV-6B in 2.7 percent. HHV-7 DNA was revealed in 12.6 percent of the studied cases. Multiple infections caused by HHV-6A and 7 were found in 2.1 percent of the samples. No statistical differences were observed regarding age, but for HHV-7 infection, an upward trend was observed in female patients. Compared to studies from other countries, low prevalence rates of herpesvirus DNA were detected in saliva from the healthy individuals in our sample. PCR methodology thus proved to be a useful tool for Roseolovirus detection and it is important to consider possible geographic and populations differences that could explain the comparatively low prevalence rates described here.

Investigation of measles IgM-seropositive cases of febrile rash illnesses in the absence of documented measles virus transmission, State of São Paulo, Brazil, 2000-2004 / Investigação dos casos de doenças febris exantemáticas com IgM reagente contra o sarampo na ausência de transmissão documentada do vírus do sarampo, Estado de São Paulo, 2000-2004

INTRODUCTION: To review measles IgM-positive cases of febrile rash illnesses in the State of São Paulo, Brazil, over the five-year period following interruption of measles virus transmission. METHODS: We reviewed 463 measles IgM-positive cases of febrile rash illness in the State of São Paulo, from 2000 to 2004. Individuals vaccinated against measles < 56 days prior to specimen collection were considered to be exposed to the vaccine. Serum from the acute and convalescent phases was tested for evidence of measles, rubella, parvovirus B19 and human herpes virus-6 infection. In the absence of seroconversion to measles immunoglobulin-G, measles IgM-positive cases were considered false positives in individuals with evidence of other viral infections. RESULTS: Among the 463 individuals with febrile rash illness who tested positive for measles IgM antibodies during the period, 297 (64 percent) were classified as exposed to the vaccine. Among the 166 cases that were not exposed to the vaccine, 109 (66 percent) were considered false positives based on the absence of seroconversion, among which 21 (13 percent) had evidence of rubella virus infection, 49 (30 percent) parvovirus B19 and 28 (17 percent) human herpes virus-6 infection. CONCLUSIONS: Following the interruption of measles virus transmission, thorough investigation of measles IgM-positive cases is required, especially among cases not exposed to the vaccine. Laboratory testing for etiologies of febrile rash illness aids interpretation of these cases.

INTRODUÇÃO: Revisar os casos de doenças febris exantemáticas com IgM reagente contra o sarampo, no Estado de São Paulo, Brasil, durante os cinco anos seguidos a interrupção da transmissão do vírus do sarampo. MÉTODOS: Nós revisamos 463 casos de doenças febris exantemáticas com IgM reagente contra o sarampo, no Estado de São Paulo, Brasil, de 2000 a 2004. Indivíduos vacinados contra o sarampo 56 dias antes da coleta de amostra foram considerados expostos à vacina. Soros da fase aguda e de convalescença foram testados para a evidência de infecção de sarampo, rubéola, parvovírus B19 e herpes vírus 6. Na ausência de soroconversão para imunoglobulina G contra o sarampo, casos com IgM reagente contra o sarampo foram considerados falsos positivos em pessoas com evidência de outras infecções virais. RESULTADOS: Entre as 463 pessoas com doenças febris exantemáticas que testaram positivo para anticorpos IgM contra o sarampo durante o período, 297 (64 por cento) pessoas foram classificadas como expostas à vacina. Entre os 166 casos não expostos à vacina, 109 (66 por cento) foram considerados falsos positivos baseado na ausência de soroconversão, dos quais 21 (13 por cento) tiveram evidência de infecção por vírus da rubéola, 49 (30 por cento) parvovírus B19 e 28 (17 por cento) infecção por herpes vírus humano 6. CONCLUSÕES: Após a interrupção da transmissão do vírus do sarampo é necessária exaustiva investigação dos casos com IgM reagente contra o sarampo, especialmente dos casos não expostos à vacina. Testes laboratoriais para etiologias das doenças febris exantemáticas ajudam na interpretação destes casos.

Epidemiology of herpes human virus 6 and epstein-bar virus infections in salivary gland neoplasms in Isfahan, Iran

Viral infections were reported to be the cause of some human malignancies. The exclusive presence of EBV [Epstein-Bar virus] and HHV6 [Herpes Human Virus 6] has been investigated in previous studies. As such comparisons had never been carried out on salivary gland neoplasms, this study aimed to determine any relationship between these two viruses in salivary gland neoplasms. Seventy eight formalin-fixed paraffin embedded tissue samples of salivary gland tumors were enrolled. The enrolled patients were those who referred to the Department of Oral Pathology of Isfahan University of Medical Sciences and to the state hospitals and private clinics in Isfahan, Iran from May 1995 to July 2005. The paraffin blocks were investigated for presence of HHV6 and EBV genomes by PCR. Out of the 78 samples, 15 were positive for both EBV and HHV6 infections while 6 were only positive for EBV, 21 were HHV6 positive but negative for EBV and 36 samples were reported negative for both viruses. A relationship was visible between EBV and HHV6 genomes. The significant relationship between HHV6 and EBV genomes and salivary gland neoplasms denotes to the question that should be answered in the light of further research whether HHV6 infection in salivary gland tumors can increase the incidence of EBV infection

Detection of human herpesvirus-7 by qualitative nested-PCR: comparison between healthy individuals and liver transplant recipients / Detecção de herpesvirus humano-7 por nested-PCR qualitativo: comparação entre indivíduos sadios e receptores de transplante hepático

Diagnosis of human herpesvirus-7 active infection in transplant patients has proved difficult, because this virus is ubiquitous and can cause persistent infections in the host. The significance of viral DNA detected in leukocytes by PCR is unclear and cross-reaction in serological tests may occur. This study aimed to evaluate nested-PCR to detect human herpesvirus-7 active infection in liver transplant recipients compared to healthy individuals. human herpesvirus-7 nested-PCR was performed on leukocytes and sera of 53 healthy volunteers and sera of 29 liver transplant recipients. In healthy volunteers, human herpesvirus-7 was detected in 28.3 percent of leukocytes and 0 percent of serum. human herpesvirus-7 was detected in sera of 48.2 percent of the liver transplant recipients. Nested-PCR on DNA extracted from leukocytes detected latent infection and the study suggests that nested-PCR performed on serum could be useful to detect human herpesvirus-7 active infection in liver transplant recipients.

Diagnóstico da infecção ativa pelo herpesvirus humano-7 é difícil devido ao fato deste vírus ser ubíquo e poder causar infecção persistente no hospedeiro. O significado da detecção do DNA viral por reação em cadeia da polimerase não é claro e, reações cruzadas podem ocorrer em testes sorológicos. O objetivo deste estudo foi avaliar a nested-PCR para detectar infecção ativa pelo herpesvirus-7 em receptores hepáticos comparando com indivíduos sadios. Nested-PCR para herpesvirus-7 foi realizado em leucócitos e soro de 53 voluntários sadios e em soro de 29 receptores hepáticos. Nos voluntários sadios, herpesvirus-7 foi detectado em 28,3 por cento de leucócitos e 0 por cento de soro. herpesvirus-7 foi detectado em soro de 48,2 por cento de receptores hepáticos. Nested-PCR em DNA extraído de leucócitos detectou infecção latente e o estudo sugere que nested-PCR realizada em soro poderia ser útil para detectar infecção ativa por herpesvirus-7 em receptores de fígado.

Incidence of human herpes virus-6 and human cytomegalovirus infections in donated bone marrow and umbilical cord blood hematopoietic stem cells.

This study examined the incidence of human herpes virus-6 (HHV-6) and human cytomegalovirus (HCMV) infections that are potentially transmitted to haematopoietic stem cells (HSC) transplant recipients via bone marrow (BM) or umbilical cord blood (UCB). Bone marrow progenitor cells were collected from 30 allogenic BM donors. UCB HSC were collected from 34 subjects. The extracted DNA was then processed using nested polymerase chain reaction (nPCR) technique. HCMV and HHV-6 serological status were determined by enzyme immunoassay (EIA). Nested PCR identified HCMV in 22 (73%) of 30 samples of BM progenitor cells but in only eight (23.5%) of 34 samples of UBC HSC ( P = 0.001). HHV-6 DNA was detected in 11 (36.6%) of 30 BM progenitor cells and in only one (2.9%) of 34 UBC cells ( P = 0.002). Both HHV-6 and HCMV infections were determined in nine (26.5%) of 34 bone marrow samples. The results indicate that, the risk of HCMV and HHV-6 via BM progenitor cells is higher than transmission by UCB cells ( P= 0.04).

Prevalence of human herpesvirus-6 in allogeneic hematopoietic stem cell transplant recipients in correlation with cytomegalovirus infection / 中国实验血液学杂志

In order to study the prevalence of human herpesvirus 6 (HHV-6) in allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients in China and to analyze the relationship between HHV-6 and cytomegalovirus (CMV) infection in post-HSCT patients, nested polymerase chain reaction (PCR) was used to monitor HHV-6 DNAemia in 72 consecutive allo-HSCT recipients. 680 EDTA anticoagulated peripheral blood specimens were gathered before HSCT or weekly until 12 weeks after allo-HSCT. The variants of HHV-6 were identified by Hind III restriction analysis. CMV-pp65 antigenemia was detected by immunofluorescence stain. The results showed that HHV-6 DNAemia was detected at least once in 62.5% (45/72) of the patients on the median day 14 (range, 7 - 63 days) after allo-HSCT, and HHV-6B was the predominant variant. CMV antigenemia was detected at least once in 65.3% (47/72) of the patients on the median day 43 (range, 14 - 105 days) after allo-HSCT. Co-infection of HHV-6 and CMV (HHV-6+/CMV+) occurred in 52.8% (38/72) recipients. The onset of HHV-6 DNAemia was earlier than that of CMV antigenemia (P < 0.0001). Patients with HHV-6 DNAemia positive were more likely to have concurrent CMV antigenemia than HHV-6 DNAemia negative patients (84.4% vs 33.3%, P = 0.0001) after allo-HSCT. Among the herpesvirus related disease, the relatively high incidence of hemorrhage cystitis (HC) occurred in 23.6% (17/72) of post-HSCT patients. 88.2% (15/17) of HC developed in HHV-6 positive patients, and 82.3% (14/17) occurred in CMV+/HHV-6+ patients. It is concluded that infection of HHV-6, co-infection of HHV-6 and CMV, commonly occurred in post-HSCT patients in China, HHV-6 infection closely related to CMV antigenemia.

Correlation between human herpesvirus 6 activation and acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the potential relationship between HHV-6 activation and acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic cell transplantation (HSCT).</p><p><b>METHODS</b>Peripheral blood samples were collected before and weekly after HSCT from 72 consecutive recipients. HHV-6 DNAemia was monitored by nested polymerase chain reaction (PCR). The genotypes of HHV-6 were identified by Hind III restriction assay.</p><p><b>RESULTS</b>Of the 72 patients, HHV-6 DNAemia were detected in 45 (62.5%) on a median of day 14 (range, 7 - 63 days) after HSCT. Grade I - IV aGHVD occurred in 40 (55.6%) on a median of day 26 (range, 9 -73 days). The median onset time of HHV-6 DNAemia was significantly earlier than that of aGHVD (P = 0.018). Compared with that in HHV-6 DNAemia negative [HHV-6(-)] patients, the cumulative incidence of grade I - IV aGHVD was higher (68.9% vs. 33.3% , P = 0.003) in HHV-6 (+) patients. Cumulative incidence of grade II - IV aGVHD in HHV-6 (+) cohort was also higher than that in HHV-6 (-) cohort (35.6% vs 14.8% , P = 0.027). Cumulative incidence of grade I - IV aGVHD was higher in patients with both HHV-6 and CMV positive (CMV+/HHV-6+) than in those with either CMV (CMV+/HHV-6-) or HHV-6 positive (CMV+/HHV-6+) and neither of them positive (CMV-/HHV-6-) [78.9% (30/38), 55. 6% (5/9) , 14. 3% (1/7) and 22. 2% (4/18), respectively, P = 0. 0001]. Cumulative incidence of grade II - IV aGVHD in CMV+/HHV-6+ group was also higher than that in CMV+/HHV-6-, CMV-/HHV-6+ and CMV-/HHV-6- groups [42.1% (16/38), 22.2% (2/9), 0% (0/7) and 11.1% (2/18), P = 0. 008].</p><p><b>CONCLUSIONS</b>Patients with HHV-6 activation or HHV-6/CMV co-infection maybe involved in the occurrence of aGVHD after HSCT.</p>

Establishment and application of fluorescent quantitative PCR method for detecting human herpes virus type 6 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish fluorescent quantitative PCR method for detecting human herpes virus type 6 (HHV6).</p><p><b>METHODS</b>According to the specific sequence of human herpes virus type 6 genes, the primers and the fluorescent probe (TaqMan) were designed and synthesized. The fragment generated from HHV 6 gene as template was cloned into the pMD18-T vector which was constructed from the pUC 18. And the positive recombinant plasmid was 1:10 diluted and used as standard quantitative template to make the standard curve for sample detection.</p><p><b>RESULTS</b>The standard curve indicated the linear relationship between Ct (cycle threshold) and template concentration. The clinical samples from 135 cases were detected by this system, 16 cases among 135 were positive.</p><p><b>CONCLUSION</b>The fluorescent quantitative PCR method for the detection of human herpes virus type 6 is simple and accurate, and this method may be helpful to clinical diagnosis.</p>

Prevalence of cytomegalovirus, human herpesvirus-6, and Epstein-Barr virus in periodontitis patients and healthy subjects in the Thai population.

Fifty periodontitis patients and 30 healthy patients with oral cavities were selected from the Faculty of Dentistry, Mahidol University, Bangkok, Thailand, from March 2001 to November 2002. Their ages varied between 15 and 70 years. Among the periodontitis patients, specimens were collected from both disease and healthy sites. All samples were evaluated for the presence of CMV, HHV-6, and EBV-1 by nested PCR. Among the periodontitis patients, CMV was found in 34%, of which 8% were at the disease sites, 10% were at the healthy sites, and 16% were from both sites. EBV was not found in this group of the patients, while HHV-6 was found in 4%, at the disease sites only. CMV was found in one (3.3%) healthy control while HHV-6 and EBV-1 were not found. The depth of sample sites, various demographic and baseline characteristics eg sex, age, occupation and root planning were not associated with the presence of these viruses.

Human herpesvirus-7 as a cause of exanthematous ilnesses in Belém, Pará, Brazil

Examinamos soros de 370 pacientes acometidos de doença exantemática, selecionados em Belém, norte do Brasil, com o propósito de se detectarem anticorpos IgM e IgG para o herpesvírus humano-7 (HHV-7). As amostras foram obtidas entre janeiro de 1996 e dezembro de 2002 e, posteriormente, processadas utilizando-se a técnica da imunofluorescência indireta (IFI). Taxas de anticorpos IgM e/ou IgG foram encontradas em 190 (51,4) desses pacientes. Observamos taxas de prevalência similares para os sexos feminino e masculino com: 52,5 e 50,3, respectivamente. O "status" sorológico foi definido pela presença de anticorpos IgG nos espécimes de 135 (36,5) pacientes. A par disso, em 55 (14,9) dos 370 pacientes foram detectados anticorpos IgM para o HHV-7. Taxas de anticorpos IgM e IgG para o HHV-7 foram similares (p > 0.05) quando comparamos indivíduos do sexo feminino e masculino: 14,4 versus 15,3 e 38,1 versus 35,0, respectivamente. Diferença estatisticamente significativa (p = 0,003) foi observada quando comparamos as taxas de anticorpos IgM para o HHV-7 nos indivíduos do grupo etário de 5-8 meses pertencentes ao sexo feminino e masculino. Taxas de prevalência variando de 4,6 (masculino, 5-8 meses de idade) a 93,3 (feminino, > 10 anos de idade) e 12,2 (masculino, 5-8 meses de idade) a 80,0 (masculino, 8-10 anos de idade) foram observadas no subgrupo positivo para IgG. Um subgrupo (n = 131) de pacientes com anticorpos IgM ou IgG foi examinado quanto a presença de DNA para o HHV-7 pela técnica da reação em cadeia da polimerase/ "nested" PCR. Infecção recente/ativa para o HHV-7 ocorreu em 11,0 (6/55) dos pacientes cujas amostras apresentaram anticorpos IgM específicos para o HHV-7. Em um subgrupo (n = 76) de pacientes com altos níveis de anticorpos IgG para o HHV-7 (título > 1: 160) não foi detectada a presença de DNA em seus soros pelo PCR/ "nested" PCR. Entre as seis infecções recentes/ativas, quatro indivíduos com menos de um ano e dois com 3 e 6 anos de idade apresentaram típico exantema súbito (E.S) definido por febre elevada (> 38,0 ºC) com duração de 24 a 72 horas, acompanhando-se de erupção cutânea maculopapular. Nossos resultados ressaltam a necessidade de procurarmos a infecção pelo HHV-7 em pacientes portadores de doença exantemática, particularmente naquelas apresentações típicas de E.S. O HHV-7 parece emergir como um novo patógeno associado a quadros exantemáticos em nossa região.