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1.
J Clin Pathol ; 73(1): 30-34, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31315894

RESUMO

AIMS: The purpose of the present study was to elucidate the presence of human herpesvirus 6A (HHV-6A), HHV-6B and HHV-7 in samples of the uterine cervix through detection of viral DNA. We analysed normal tissues, samples with low-grade squamous intraepithelial lesions (LSILs) and high-grade squamous intraepithelial lesions (HSILs). We correlated the presence of HHV-6 and HHV-7 with the finding of human papillomavirus (HPV) in mucosal samples. METHODS: Cervical samples were examined and grouped as follows: group 1 (n=29), normal cytology; group 2 (n=61), samples with LSIL; group 3 (n=35), samples with HSIL. Molecular biology examinations were performed in all samples to detect HHV-6, HHV-7 and HPV DNA and to typify HHV-6 species. RESULTS: Group 1: normal cytology and HPV (-): HHV-6: 6.8% (2/29), HHV-7: 79.3% (23/29); group 2: LSIL and HPV (-): HHV-6: 93.1% (27/29), HHV-7: 96.5% (28/29); LSIL and HPV (+): HHV-6: 0% (0/32), HHV-7: 90.6% (29/32); group 3: HSIL and HPV (-): HHV-6: 20% (2/10), HHV-7: 70% (7/10); HSIL HPV (+): HHV-6: 12% (3/25), HHV-7: 68% (17/25). HHV-6A DNA was not detected in any samples. CONCLUSIONS: (1) Both HHV-6 and HHV-7 infect the mucosal cells of the cervix with higher prevalence of HHV-7. (2) The higher prevalence of HHV-6 in LSIL HPV (-) samples compared with those with normal cytology indicates that it constitutes a possible risk factor for atypia production. (3) The presence of HHV-7 in all samples questions its role in the production of atypia. (4) The finding of HHV-6 and HHV-7 suggests that the cervical mucosa is a possible transmission pathway for these viruses.


Assuntos
Neoplasia Intraepitelial Cervical/diagnóstico , DNA Viral/genética , Herpesvirus Humano 6/genética , Herpesvirus Humano 7/genética , Técnicas de Diagnóstico Molecular , Infecções por Roseolovirus/diagnóstico , Lesões Intraepiteliais Escamosas Cervicais/diagnóstico , Neoplasias do Colo do Útero/diagnóstico , Adolescente , Adulto , Argentina , Neoplasia Intraepitelial Cervical/genética , Neoplasia Intraepitelial Cervical/virologia , Feminino , Testes de DNA para Papilomavírus Humano , Humanos , Pessoa de Meia-Idade , Papillomaviridae/genética , Infecções por Papillomavirus/diagnóstico , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/virologia , Valor Preditivo dos Testes , Prognóstico , Estudos Retrospectivos , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/transmissão , Infecções por Roseolovirus/virologia , Lesões Intraepiteliais Escamosas Cervicais/genética , Lesões Intraepiteliais Escamosas Cervicais/virologia , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/virologia , Adulto Jovem
2.
J Virol ; 93(3)2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30429336

RESUMO

Human herpesvirus 6B (HHV-6B) DNA is frequently detected in human samples. Diagnostic assays distinguishing HHV-6B reactivation from latency are limited. This has impaired strategies to diagnose and treat HHV-6B-associated diseases. We used RNA sequencing to characterize and compare the HHV-6B transcriptome in multiple sample types, including (i) whole blood from hematopoietic cell transplant (HCT) recipients with and without HHV-6B plasma viremia, (ii) tumor tissue samples from subjects with large B cell lymphoma infected with HHV-6B, (iii) lymphoblastoid cell lines (LCLs) from subjects with inherited chromosomally integrated HHV-6B or latent infection with HHV-6B, and (iv) HHV-6B Z29 infected SupT1 CD4+ T cells. We demonstrated substantial overlap in the HHV-6B transcriptome observed in in vivo and in vitro samples, although there was variability in the breadth and quantity of gene expression across samples. The HHV-6B viral polymerase gene U38 was the only HHV-6B transcript detected in all next-generation RNA sequencing (RNA-seq) data sets and was one of the most highly expressed genes. We developed a novel reverse transcription-PCR assay targeting HHV-6B U38, which identified U38 mRNA in all tested whole-blood samples from patients with concurrent HHV-6B viremia. No HHV-6B U38 transcripts were detected by RNA-seq or reverse transcription-real-time quantitative PCR (RT-qPCR) in whole-blood samples from subjects without HHV-6B plasma detection or from latently infected LCLs. A RT-qPCR assay for HHV-6B U38 may be useful to identify lytic HHV-6B infection in nonplasma samples and samples from individuals with inherited chromosomally integrated HHV-6B. This study also demonstrates the feasibility of transcriptomic analyses for HCT recipients.IMPORTANCE Human herpesvirus 6B (HHV-6B) is a DNA virus that infects most children within the first few years of life. After primary infection, HHV-6B persists as a chronic, latent infection in many cell types. Additionally, HHV-6B can integrate into germ line chromosomes, resulting in individuals with viral DNA in every nucleated cell. Given that PCR to detect viral DNA is the mainstay for diagnosing HHV-6B infection, the characteristics of HHV-6B infection complicate efforts to distinguish between latent and active viral infection, particularly in immunocompromised patients who have frequent HHV-6B reactivation. In this study, we used RNA sequencing to characterize the HHV-6B gene expression profile in multiple sample types, and our findings identified evidence-based targets for diagnostic tests that distinguish between latent and active viral infection.


Assuntos
Herpesvirus Humano 6/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Infecções por Roseolovirus/diagnóstico , Transcriptoma , Proteínas Virais/genética , Viremia/diagnóstico , Ativação Viral , Latência Viral , Adulto , Idoso , Biomarcadores/análise , Estudos de Casos e Controles , Citocinas/sangue , DNA Viral , Feminino , Humanos , Linfoma Difuso de Grandes Células B/genética , Linfoma Difuso de Grandes Células B/patologia , Linfoma Difuso de Grandes Células B/virologia , Masculino , Pessoa de Meia-Idade , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/virologia , Viremia/genética , Viremia/virologia
3.
Front Immunol ; 9: 2803, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30574140

RESUMO

The aberrant expression of human endogenous retrovirus (HERV) elements of the HERV-W family has been associated with different diseases, including multiple sclerosis (MS). In particular, the expression of the envelope protein (Env) from the multiple sclerosis-associated retrovirus (MSRV), a member of HERV-W family and known for its potent proinflammatory activity, is repeatedly detected in the brain lesions and blood of MS patients. Furthermore, human herpesvirus 6 (HHV-6) infection has long been suspected to play a role in the pathogenesis of MS and neuroinflammation. We show here that both HHV-6A and stimulation of its receptor, transmembrane glycoprotein CD46, induce the expression of MSRV-Env. The engagement of extracellular domains SCR3 and SCR4 of CD46-Cyt1 isoform was required for MSRV-env transactivation, limiting thus the MSRV-Env induction to the CD46 ligands binding these domains, including C3b component of complement, specific monoclonal antibodies, and both infectious and UV-inactivated HHV-6A, but neither HHV-6B nor measles virus vaccine strain. Induction of MSRV-Env required CD46 Cyt-1 singling and was abolished by the inhibitors of protein kinase C. Finally, both membrane-expressed and secreted MSRV-Env trigger TLR4 signaling, displaying thus a proinflammatory potential, characteristic for this viral protein. These data expand the specter of HHV-6A effects in the modulation of the immune response and support the hypothesis that cross-talks between exogenous and endogenous viruses may contribute to inflammatory diseases and participate in neuroinflammation. Furthermore, they reveal a new function of CD46, known as an inhibitor of complement activation and receptor for several pathogens, in transactivation of HERV env genes, which may play an important role in the pathogenesis of inflammatory diseases.


Assuntos
Retrovirus Endógenos , Herpesvirus Humano 6 , Proteína Cofatora de Membrana , Esclerose Múltipla , Proteínas da Gravidez , Infecções por Roseolovirus , Linhagem Celular Tumoral , Retrovirus Endógenos/genética , Retrovirus Endógenos/imunologia , Retrovirus Endógenos/metabolismo , Herpesvirus Humano 6/imunologia , Herpesvirus Humano 6/metabolismo , Humanos , Inflamação/genética , Inflamação/imunologia , Inflamação/patologia , Inflamação/virologia , Proteína Cofatora de Membrana/imunologia , Proteína Cofatora de Membrana/metabolismo , Esclerose Múltipla/genética , Esclerose Múltipla/imunologia , Esclerose Múltipla/metabolismo , Esclerose Múltipla/virologia , Proteínas da Gravidez/biossíntese , Proteínas da Gravidez/genética , Proteínas da Gravidez/imunologia , Domínios Proteicos , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/imunologia , Infecções por Roseolovirus/metabolismo
4.
Int J Hematol ; 108(5): 535-542, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30014227

RESUMO

In this prospective observational study, we compared the human herpesvirus-6 (HHV-6) DNA load in serially collected paired plasma and whole blood (WB) samples from allogeneic hematopoietic stem cell transplantation (HSCT) recipients. A total of 721 paired samples were collected from 68 recipients. The positive rate for HHV-6 DNA was 9.7 and 35.0% in plasma and WB samples, respectively (P < 0.001). The correlation of HHV-6 DNA load between plasma and WB was poor (R2 = 0.250). After reaching peak levels, HHV-6 DNA showed a delayed decrease in WB in comparison with plasma (median, 28 versus 7 days, P < 0.001). We additionally tested HHV-6 mRNA status in 95 samples from eight patients. To identify positive HHV-6 mRNA, plasma HHV-6 DNA showed 55.0% sensitivity and 100% specificity, whereas WB HHV-6 DNA showed 90.0% sensitivity and 68.0% specificity. The false-positive rate for identifying positive HHV-6 mRNA was 0% for plasma HHV-6 DNA and 32.0% for WB HHV-6 DNA. Although WB was more sensitive than plasma for detecting HHV-6 reactivation, the rates of false positivity for active HHV-6 infection were higher for WB than for plasma.


Assuntos
DNA Viral/sangue , Neoplasias Hematológicas/sangue , Neoplasias Hematológicas/terapia , Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 6 , Infecções por Roseolovirus/sangue , Adolescente , Adulto , Aloenxertos , DNA Viral/genética , Reações Falso-Positivas , Feminino , Neoplasias Hematológicas/genética , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Estudos Prospectivos , Infecções por Roseolovirus/etiologia , Infecções por Roseolovirus/genética
5.
Neuron ; 99(1): 64-82.e7, 2018 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-29937276

RESUMO

Investigators have long suspected that pathogenic microbes might contribute to the onset and progression of Alzheimer's disease (AD) although definitive evidence has not been presented. Whether such findings represent a causal contribution, or reflect opportunistic passengers of neurodegeneration, is also difficult to resolve. We constructed multiscale networks of the late-onset AD-associated virome, integrating genomic, transcriptomic, proteomic, and histopathological data across four brain regions from human post-mortem tissue. We observed increased human herpesvirus 6A (HHV-6A) and human herpesvirus 7 (HHV-7) from subjects with AD compared with controls. These results were replicated in two additional, independent and geographically dispersed cohorts. We observed regulatory relationships linking viral abundance and modulators of APP metabolism, including induction of APBB2, APPBP2, BIN1, BACE1, CLU, PICALM, and PSEN1 by HHV-6A. This study elucidates networks linking molecular, clinical, and neuropathological features with viral activity and is consistent with viral activity constituting a general feature of AD.


Assuntos
Doença de Alzheimer/virologia , Precursor de Proteína beta-Amiloide/metabolismo , Encéfalo/virologia , Encefalite Viral/virologia , Herpesvirus Humano 6 , Herpesvirus Humano 7 , Infecções por Roseolovirus/virologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Doença de Alzheimer/patologia , Secretases da Proteína Precursora do Amiloide/genética , Animais , Ácido Aspártico Endopeptidases/genética , Encéfalo/metabolismo , Encéfalo/patologia , Estudos de Casos e Controles , Clusterina/genética , Estudos de Coortes , Encefalite Viral/genética , Encefalite Viral/metabolismo , Encefalite Viral/patologia , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Genômica , Humanos , Camundongos , Camundongos Knockout , Camundongos Transgênicos , MicroRNAs/genética , Microbiota , Proteínas Monoméricas de Montagem de Clatrina/genética , Proteínas Nucleares/genética , Presenilina-1/genética , Proteômica , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/metabolismo , Infecções por Roseolovirus/patologia , Proteínas Supressoras de Tumor/genética , Carga Viral
6.
J Virol ; 92(10)2018 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-29491155

RESUMO

Quantitative PCR is a diagnostic pillar for clinical virology testing, and reference materials are necessary for accurate, comparable quantitation between clinical laboratories. Accurate quantitation of human herpesvirus 6A/B (HHV-6A/B) is important for detection of viral reactivation and inherited chromosomally integrated HHV-6A/B in immunocompromised patients. Reference materials in clinical virology commonly consist of laboratory-adapted viral strains that may be affected by the culture process. We performed next-generation sequencing to make relative copy number measurements at single nucleotide resolution of eight candidate HHV-6A and seven HHV-6B reference strains and DNA materials from the HHV-6 Foundation and Advanced Biotechnologies Inc. Eleven of 17 (65%) HHV-6A/B candidate reference materials showed multiple copies of the origin of replication upstream of the U41 gene by next-generation sequencing. These large tandem repeats arose independently in culture-adapted HHV-6A and HHV-6B strains, measuring 1,254 bp and 983 bp, respectively. The average copy number measured was between 5 and 10 times the number of copies of the rest of the genome. We also report the first interspecies recombinant HHV-6A/B strain with a HHV-6A backbone and a >5.5-kb region from HHV-6B, from U41 to U43, that covered the origin tandem repeat. Specific HHV-6A reference strains demonstrated duplication of regions at U1/U2, U87, and U89, as well as deletion in the U12-to-U24 region and the U94/U95 genes. HHV-6A/B strains derived from cord blood mononuclear cells from different laboratories on different continents with fewer passages revealed no copy number differences throughout the viral genome. These data indicate that large origin tandem duplications are an adaptation of both HHV-6A and HHV-6B in culture and show interspecies recombination is possible within the Betaherpesvirinae.IMPORTANCE Anything in science that needs to be quantitated requires a standard unit of measurement. This includes viruses, for which quantitation increasingly determines definitions of pathology and guidelines for treatment. However, the act of making standard or reference material in virology can alter its very accuracy through genomic duplications, insertions, and rearrangements. We used deep sequencing to examine candidate reference strains for HHV-6, a ubiquitous human virus that can reactivate in the immunocompromised population and is integrated into the human genome in every cell of the body for 1% of people worldwide. We found large tandem repeats in the origin of replication for both HHV-6A and HHV-6B that are selected for in culture. We also found the first interspecies recombinant between HHV-6A and HHV-6B, a phenomenon that is well known in alphaherpesviruses but to date has not been seen in betaherpesviruses. These data critically inform HHV-6A/B biology and the standard selection process.


Assuntos
Variações do Número de Cópias de DNA/genética , Herpesvirus Humano 6/genética , Origem de Replicação/genética , Sequências de Repetição em Tandem/genética , Sequência de Bases , Linhagem Celular , DNA Viral/genética , Genoma Viral/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/virologia , Análise de Sequência de DNA
7.
Med Hypotheses ; 112: 47-50, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29447938

RESUMO

Human Herpes Virus type 6 (HHV-6) is a ubiquitous virus consisting of two viral species, HHV-6A and HHV-6B that have been associated with numerous and diverse pathologies. As many other viruses HHV-6 modulates the apoptotic machinery of its host to subvert immune response to infection, yet the exact mechanisms behind this process remain under investigation. The genes encoding the CTSS, PTX3, CHI3L1, Mx1, CXCL16, BIRC3 and BST2 proteins have been linked to HHV-6Α related neurologic diseases whilst also associated with apoptosis. This study aimed at the identification and functional analysis of the gene interaction network (interactome) of CTSS-PTX3-CHI3L1-Mx1-CXCL16-BIRC3-BST2 so as to evaluate the hypothesis of a probable link between the latter and host's immune response to HHV-6A infection.


Assuntos
Imunidade Adaptativa/genética , Proteínas Reguladoras de Apoptose/genética , Redes Reguladoras de Genes , Herpesvirus Humano 6/fisiologia , Interações Hospedeiro-Patógeno/imunologia , Modelos Genéticos , Modelos Imunológicos , Infecções por Roseolovirus/imunologia , Antígenos CD/biossíntese , Antígenos CD/genética , Proteínas Reguladoras de Apoptose/biossíntese , Proteína 3 com Repetições IAP de Baculovírus/biossíntese , Proteína 3 com Repetições IAP de Baculovírus/genética , Proteína C-Reativa/biossíntese , Proteína C-Reativa/genética , Catepsinas/biossíntese , Catepsinas/genética , Quimiocina CXCL16/biossíntese , Quimiocina CXCL16/genética , Proteína 1 Semelhante à Quitinase-3/biossíntese , Proteína 1 Semelhante à Quitinase-3/genética , Proteínas Ligadas por GPI/biossíntese , Proteínas Ligadas por GPI/genética , Regulação Viral da Expressão Gênica/imunologia , Ontologia Genética , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/imunologia , Interações Hospedeiro-Patógeno/genética , Humanos , Evasão da Resposta Imune/genética , Proteínas de Resistência a Myxovirus/biossíntese , Proteínas de Resistência a Myxovirus/genética , Infecções por Roseolovirus/genética , Componente Amiloide P Sérico/biossíntese , Componente Amiloide P Sérico/genética
8.
Sci Rep ; 8(1): 3472, 2018 02 22.
Artigo em Inglês | MEDLINE | ID: mdl-29472617

RESUMO

Human herpesviruses 6-A and -B (HHV-6A, HHV-6B) are ubiquitous in human populations worldwide. These viruses have been associated with several diseases such as multiple sclerosis, Hodgkin's lymphoma or encephalitis. Despite of the need to understand the genetic diversity and geographic stratification of these viruses, the availability of complete viral sequences from different populations is still limited. Here, we present nine new inherited chromosomally integrated HHV-6 sequences from diverse geographical origin which were generated through target DNA enrichment on lymphoblastoid cell lines derived from healthy individuals. Integration with available HHV-6 sequences allowed the assessment of HHV-6A and -6B phylogeny, patterns of recombination and signatures of natural selection. Analysis of the intra-species variability showed differences between A and B diversity levels and revealed that the HHV-6B reference (Z29) is an uncommon sequence, suggesting the need for an alternative reference sequence. Signs of geographical variation are present and more defined in HHV-6A, while they appear partly masked by recombination in HHV-6B. Finally, we conducted a scan for signatures of selection in protein coding genes that yielded at least 6 genes (4 and 2 respectively for the A and B species) showing significant evidence for accelerated evolution, and 1 gene showing evidence of positive selection in HHV-6A.


Assuntos
Variação Genética , Herpesvirus Humano 6/genética , Infecções por Roseolovirus/epidemiologia , Infecções por Roseolovirus/virologia , Linhagem Celular , DNA Viral/genética , Geografia , Herpesvirus Humano 6/patogenicidade , Humanos , Filogeografia , Infecções por Roseolovirus/genética , Integração Viral/genética
9.
Klin Lab Diagn ; 63(6): 357-361, 2018.
Artigo em Russo | MEDLINE | ID: mdl-30702227

RESUMO

The purpose of the article is to reveal the immunological features of the infection caused by the herpesvirus type 6 virus, based on the level of the polymorphism IFNG geneγ (+874 A / T) and interferon production. Group I (n = 58) consisted of children with herpetic infection of type 6, severe form. Group II (n = 31) included children with herpetic infection of type 6, medium-heavy form. The control group consisted of 53 healthy newborns born. The allelic variants of the genes were determined by the PCR method followed by restriction analysis of the GosNII Genetics test systems (Moscow) Among the allelic variants of the interferon γ gene, patients with a severe form of herpetic infection of type 6 are significantly more likely than in children with a moderate-to-severe course of the disease, and the AA genotype of the polymorphism +874 of the IFNG gene is detected in the control group and significantly less than the control group, the allele T and the TT genotype. The level of interferon-γ had a significant decrease in children with severe infection in comparison with the control group. Exposure of the infection is associated with the A allele and the AA genotype of the polymorphic region +874 A / T of the IFNG gene.


Assuntos
Predisposição Genética para Doença , Interferon gama/genética , Polimorfismo de Nucleotídeo Único , Infecções por Roseolovirus/genética , Alelos , Estudos de Casos e Controles , Genótipo , Herpesvirus Humano 6 , Humanos , Recém-Nascido
10.
J Immunol ; 199(9): 3212-3221, 2017 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-28972091

RESUMO

A recently described mouse homolog of the human roseoloviruses, murine roseolovirus (MRV), causes loss of peripheral and thymic CD4+ cells during neonatal infection of BALB/c mice. Despite significant disruptions to the normal adaptive immune response, infected BALB/c mice reproducibly recover from infection, consistent with prior studies on a related virus, mouse thymic virus. In this article, we show that, in contrast to published studies on mouse thymic virus, MRV appears to robustly infect neonatal C57BL/6 (B6) mice, causing severe depletion of thymocytes and peripheral T cells. Moreover, B6 mice recovered from infection. We investigated the mechanism of thymocyte and T cell loss, determining that the major thymocyte subsets were infected with MRV; however, CD4+ and CD4+CD8- T cells showed increased apoptosis during infection. We found that CD8+ T cells populated MRV-infected thymi. These CD8+ T cells expressed markers of activation, had restricted TCR repertoire, and accumulated intracellular effector proteins, consistent with a cytotoxic lymphocyte phenotype and suggesting their involvement in viral clearance. Indeed, absence of CD8+ T cells prevented recovery from MRV infection and led to lethality in infected animals, whereas B cell-deficient mice showed CD4+ T cell loss but recovered from infection without lethality. Thus, these results demonstrate that CD8+ T cells are required for protective immunity against a naturally occurring murine pathogen that infects the thymus and establish a novel infection model for MRV in B6 mice, providing the foundation for detailed future studies on MRV with the availability of innumerable mutant mice on the B6 background.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Infecções por Roseolovirus/imunologia , Roseolovirus/imunologia , Timo/imunologia , Animais , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/patologia , Camundongos , Camundongos Knockout , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/patologia , Timo/patologia
11.
J Gen Virol ; 98(7): 1823-1830, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28699856

RESUMO

Integration of the complete human herpesvirus 6 (HHV-6) genome into the telomere of a chromosome has been reported in some individuals (inherited chromosomally integrated HHV-6; iciHHV-6). Since the proportion of iciHHV-6-positive individuals with integration in chromosome 22 is high in Japan, we hypothesized a founder effect. In this study, we sought to elucidate the reason for the high proportion of viral integrations into chromosome 22. We analyzed six cases of iciHHV-6A and two cases of iciHHV-6B, including one iciHHV-6A case with a matched sample from a father and one iciHHV-6B case with a matched sample from a mother. In iciHHV-6A, the same copy numbers of viral telomeric repeat sequences (TRS) and the same five microsatellite markers were detected in both the index case and paternal sample. Moreover, the same five microsatellite markers were demonstrated in four cases and the same copy numbers of viral TRS were demonstrated in two pairs of two cases. The present microsatellite analysis suggested that the viral genomes detected in some iciHHV-6A patients were derived from a common ancestral integration.


Assuntos
Cromossomos Humanos Par 22/virologia , Herpesvirus Humano 6/isolamento & purificação , Infecções por Roseolovirus/virologia , Adulto , Cromossomos Humanos Par 22/genética , Feminino , Genoma Viral , Herpesvirus Humano 6/classificação , Herpesvirus Humano 6/genética , Herpesvirus Humano 6/fisiologia , Humanos , Japão , Masculino , Sequências Repetitivas de Ácido Nucleico , Infecções por Roseolovirus/congênito , Infecções por Roseolovirus/genética , Integração Viral
12.
Viruses ; 10(1)2017 12 31.
Artigo em Inglês | MEDLINE | ID: mdl-29301233

RESUMO

Tissue-culture adaptation of viruses can modulate infection. Laboratory passage and bacterial artificial chromosome (BAC)mid cloning of human cytomegalovirus, HCMV, resulted in genomic deletions and rearrangements altering genes encoding the virus entry complex, which affected cellular tropism, virulence, and vaccine development. Here, we analyse these effects on the reference genome for related betaherpesviruses, Roseolovirus, human herpesvirus 6A (HHV-6A) strain U1102. This virus is also naturally "cloned" by germline subtelomeric chromosomal-integration in approximately 1% of human populations, and accurate references are key to understanding pathological relationships between exogenous and endogenous virus. Using whole genome next-generation deep-sequencing Illumina-based methods, we compared the original isolate to tissue-culture passaged and the BACmid-cloned virus. This re-defined the reference genome showing 32 corrections and 5 polymorphisms. Furthermore, minor variant analyses of passaged and BACmid virus identified emerging populations of a further 32 single nucleotide polymorphisms (SNPs) in 10 loci, half non-synonymous indicating cell-culture selection. Analyses of the BAC-virus genome showed deletion of the BAC cassette via loxP recombination removing green fluorescent protein (GFP)-based selection. As shown for HCMV culture effects, select HHV-6A SNPs mapped to genes encoding mediators of virus cellular entry, including virus envelope glycoprotein genes gB and the gH/gL complex. Comparative models suggest stabilisation of the post-fusion conformation. These SNPs are essential to consider in vaccine-design, antimicrobial-resistance, and pathogenesis.


Assuntos
Adaptação Fisiológica/genética , Herpesvirus Humano 6/genética , Sequenciamento de Nucleotídeos em Larga Escala , Infecções por Roseolovirus/virologia , Proteínas do Envelope Viral/genética , Internalização do Vírus , Sequenciamento Completo do Genoma , Linhagem Celular , Genoma Viral/genética , Herpesvirus Humano 6/fisiologia , Humanos , Modelos Moleculares , Polimorfismo de Nucleotídeo Único , Infecções por Roseolovirus/genética , Seleção Genética , Deleção de Sequência , Proteínas do Envelope Viral/química
13.
Transpl Infect Dis ; 19(1)2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27859994

RESUMO

Chromosomally integrated human herpesvirus 6 (ciHHV-6) can be transmitted via allogeneic hematopoietic cell transplantation. To date, only a few cases have been reported. Here, we report a case identified as transmission of ciHHV-6 via cord blood transplantation. Distinguishing transmission of ciHHV-6 from HHV-6 reactivation in cases with high titer of HHV-6 DNA load after transplantation is important to prevent unnecessary exposure to antiviral drugs that could be toxic.


Assuntos
Cromossomos Humanos Par 22/virologia , Transplante de Células-Tronco de Sangue do Cordão Umbilical/efeitos adversos , Sangue Fetal/virologia , Herpesvirus Humano 6/genética , Agonistas Mieloablativos/efeitos adversos , Infecções por Roseolovirus/transmissão , Condicionamento Pré-Transplante/efeitos adversos , Integração Viral , Aciclovir/administração & dosagem , Aciclovir/análogos & derivados , Aciclovir/uso terapêutico , Antibioticoprofilaxia , Antivirais/administração & dosagem , Antivirais/uso terapêutico , Bussulfano/efeitos adversos , Bussulfano/uso terapêutico , Pré-Escolar , DNA Viral/isolamento & purificação , Exantema/sangue , Exantema/virologia , Febre/sangue , Febre/virologia , Herpesvirus Humano 6/isolamento & purificação , Humanos , Hospedeiro Imunocomprometido , Masculino , Melfalan/efeitos adversos , Melfalan/uso terapêutico , Agonistas Mieloablativos/uso terapêutico , Infecções por Roseolovirus/sangue , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/virologia , Doadores não Relacionados , Valaciclovir , Valina/administração & dosagem , Valina/análogos & derivados , Valina/uso terapêutico , Carga Viral
14.
Oncotarget ; 7(30): 48070-48080, 2016 Jul 26.
Artigo em Inglês | MEDLINE | ID: mdl-27344170

RESUMO

Human Herpesvirus 6 (HHV-6) has been involved in the development of several central nervous system (CNS) diseases, such as Alzheimer's disease, multiple sclerosis and glioma. In order to identify the pathogenic mechanism of HHV-6A infection, we carried out mRNA-seq study of human astrocyte HA1800 cell with HHV-6A GS infection. Using mRNA-seq analysis of HA1800-control cells with HA1800-HHV-6A GS cells, we identified 249 differentially expressed genes. After investigating these candidate genes, we found seven genes associated with two or more CNS diseases: CTSS, PTX3, CHI3L1, Mx1, CXCL16, BIRC3, and BST2. This is the first transcriptome sequencing study which showed the significant association of these genes between HHV-6A infection and neurologic diseases. We believe that our findings can provide a new perspective to understand the pathogenic mechanism of HHV-6A infection and neurologic diseases.


Assuntos
Astrócitos/fisiologia , Astrócitos/virologia , Herpesvirus Humano 6/fisiologia , Doenças do Sistema Nervoso/genética , Doenças do Sistema Nervoso/virologia , Infecções por Roseolovirus/genética , Técnicas de Cultura de Células , Humanos , Doenças do Sistema Nervoso/patologia , Transcriptoma
15.
PLoS Pathog ; 12(5): e1005666, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27244446

RESUMO

Human herpesvirus 6A (HHV-6A) and 6B (HHV-6B) are ubiquitous betaherpesviruses that infects humans within the first years of life and establishes latency in various cell types. Both viruses can integrate their genomes into telomeres of host chromosomes in latently infected cells. The molecular mechanism of viral integration remains elusive. Intriguingly, HHV-6A, HHV-6B and several other herpesviruses harbor arrays of telomeric repeats (TMR) identical to human telomere sequences at the ends of their genomes. The HHV-6A and HHV-6B genomes harbor two TMR arrays, the perfect TMR (pTMR) and the imperfect TMR (impTMR). To determine if the TMR are involved in virus integration, we deleted both pTMR and impTMR in the HHV-6A genome. Upon reconstitution, the TMR mutant virus replicated comparable to wild type (wt) virus, indicating that the TMR are not essential for HHV-6A replication. To assess the integration properties of the recombinant viruses, we established an in vitro integration system that allows assessment of integration efficiency and genome maintenance in latently infected cells. Integration of HHV-6A was severely impaired in the absence of the TMR and the virus genome was lost rapidly, suggesting that integration is crucial for the maintenance of the virus genome. Individual deletion of the pTMR and impTMR revealed that the pTMR play the major role in HHV-6A integration, whereas the impTMR only make a minor contribution, allowing us to establish a model for HHV-6A integration. Taken together, our data shows that the HHV-6A TMR are dispensable for virus replication, but are crucial for integration and maintenance of the virus genome in latently infected cells.


Assuntos
Herpesvirus Humano 6/genética , Infecções por Roseolovirus/genética , Telômero/genética , Integração Viral/genética , DNA Viral/genética , Humanos , Reação em Cadeia da Polimerase , Replicação Viral/genética
16.
Ann Biol Clin (Paris) ; 74(2): 156-67, 2016.
Artigo em Francês | MEDLINE | ID: mdl-27029721

RESUMO

The diagnosis of the infection by the human herpesviruses 6A and 6B (HHV-6A, HHV-6B) is based on a direct and an indirect approaches. Serological methods are mainly used to ask primary infection diagnosis and carry out epidemiological studies. However, limitations are numerous with, in particular, the existence of cross-reactivity with other herpesviruses, and the inability to differentiate the two kinds of HHV-6. Initially based on virus isolation in cell culture, direct diagnosis evolved with the development of gene amplification methods that provide sensitivity and specificity, and allow viral quantitation in many biological systems and the identification of present species. Its main current indications are the identification of active infection, the identification of the integrated form of HHV-6 (iciHHV-6, inherited chromosomally integrated HHV-6) and the monitoring of the effectiveness of antiviral treatment.


Assuntos
Herpesvirus Humano 6/isolamento & purificação , Infecções por Roseolovirus/diagnóstico , Infecções por Roseolovirus/terapia , Antivirais/uso terapêutico , Monitoramento de Medicamentos/métodos , Herpesvirus Humano 6/genética , Humanos , Reação em Cadeia da Polimerase/métodos , Infecções por Roseolovirus/genética , Resultado do Tratamento
17.
Viruses ; 8(1)2016 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-26784220

RESUMO

Human herpesvirus-6A and B (HHV-6A, HHV-6B) have recently defined endogenous genomes, resulting from integration into the germline: chromosomally-integrated "CiHHV-6A/B". These affect approximately 1.0% of human populations, giving potential for virus gene expression in every cell. We previously showed that CiHHV-6A was more divergent than CiHHV-6B by examining four genes in 44 European CiHHV-6A/B cardiac/haematology patients. There was evidence for gene expression/reactivation, implying functional non-defective genomes. To further define the relationship between HHV-6A and CiHHV-6A we used next-generation sequencing to characterize genomes from three CiHHV-6A cardiac patients. Comparisons to known exogenous HHV-6A showed CiHHV-6A genomes formed a separate clade; including all 85 non-interrupted genes and necessary cis-acting signals for reactivation as infectious virus. Greater single nucleotide polymorphism (SNP) density was defined in 16 genes and the direct repeats (DR) terminal regions. Using these SNPs, deep sequencing analyses demonstrated superinfection with exogenous HHV-6A in two of the CiHHV-6A patients with recurrent cardiac disease. Characterisation of the integration sites in twelve patients identified the human chromosome 17p subtelomere as a prevalent site, which had specific repeat structures and phylogenetically related CiHHV-6A coding sequences indicating common ancestral origins. Overall CiHHV-6A genomes were similar, but distinct from known exogenous HHV-6A virus, and have the capacity to reactivate as emerging virus infections.


Assuntos
Cromossomos Humanos/virologia , Insuficiência Cardíaca/virologia , Herpesvirus Humano 6/fisiologia , Infecções por Roseolovirus/virologia , Integração Viral , Sequência de Bases , Cromossomos Humanos/genética , Insuficiência Cardíaca/genética , Herpesvirus Humano 6/genética , Humanos , Dados de Sequência Molecular , Filogenia , Estudos Prospectivos , Infecções por Roseolovirus/genética
18.
J Virol Methods ; 227: 47-9, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26542463

RESUMO

When using relative gene expression for quantification of RNA it is crucial that the reference genes used for normalization do not change with the experimental condition. We aimed at investigating the expressional stability of commonly used reference genes during Human herpesvirus 6B (HHV-6B) infection. Expression of eight commonly used reference genes were investigated with quantitative PCR in a T-cell line infected with HHV-6B. The stability of genes was investigated using the 2(-ΔΔCT) method and the algorithms BestKeeper, GeNorm and NormFinder. Our results indicate that peptidylprolyl isomerase A (PPIA) is the most stably expressed gene while TATA box binding protein (TBP) is the least stably expressed gene during HHV-6B infection. In a confirmatory experiment, TBP was demonstrated to be dose and time dependently upregulated by HHV-6B. The stability of PPIA is in line with other studies investigating different herpesvirus infections whereas the finding that HHV-6B significantly upregulates TBP is novel and most likely specific to HHV-6B.


Assuntos
Herpesvirus Humano 6/genética , Peptidilprolil Isomerase/genética , Infecções por Roseolovirus/genética , Proteína de Ligação a TATA-Box/genética , Biomarcadores , Expressão Gênica , Humanos , Peptidilprolil Isomerase/metabolismo , Infecções por Roseolovirus/enzimologia , Infecções por Roseolovirus/metabolismo , Proteína de Ligação a TATA-Box/metabolismo , Regulação para Cima
19.
Clin Microbiol Infect ; 22(2): 209.e5-209.e8, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26482270

RESUMO

To evaluate the human herpes virus 6 (HHV-6) -specific immune response in individuals with chromosomally integrated HHV-6 (ciHHV-6), we measured HHV-6-antigen-specific cytokine responses (interferon-γ, interleukin-2, tumour necrosis factor-α) in T cells by flow cytometry in 12 and 16 individuals with and without ciHHV-6, respectively. All individuals with ciHHV-6 showed HHV-6-specific T cells with higher frequencies of HHV-6-specific CD8(+) cells (0.03-14.93, median 2.15% of CD8(+) cells) compared with non-ciHHV-6 (0.0-10.67, median 0.36%, p 0.026). The observed increased HHV-6-specific functionally active responses in individuals with ciHHV-6 clearly disprove speculations on immune tolerance in ciHHV-6 and indicate clinical and immunological implications of ciHHV-6.


Assuntos
Citocinas/metabolismo , Herpesvirus Humano 6/genética , Infecções por Roseolovirus/virologia , Linfócitos T/citologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Cromossomos Humanos , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/imunologia , Linfócitos T/metabolismo , Integração Viral , Adulto Jovem
20.
Biol Blood Marrow Transplant ; 21(2): 371-3, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25255164

RESUMO

We identified 37 hematopoietic cell transplantation recipients with human herpesvirus 6 (HHV-6) central nervous system dysfunction and tested donor-recipient pairs for chromosomally integrated HHV-6 (ciHHV-6). One patient had ciHHV-6A with possible HHV-6A reactivation and encephalitis. There was no ciHHV-6 enrichment in this group, but larger studies are needed to determine if patients with ciHHV-6 are at increased risk for HHV-6-associated diseases or other complications.


Assuntos
Cromossomos Humanos/virologia , DNA Viral/líquido cefalorraquidiano , Encefalite Viral/virologia , Herpesvirus Humano 6/genética , Infecções por Roseolovirus/virologia , Integração Viral , Sistema Nervoso Central/metabolismo , Sistema Nervoso Central/patologia , Sistema Nervoso Central/virologia , Cromossomos Humanos/química , Encefalite Viral/líquido cefalorraquidiano , Encefalite Viral/genética , Encefalite Viral/patologia , Transplante de Células-Tronco Hematopoéticas , Herpesvirus Humano 6/classificação , Herpesvirus Humano 6/isolamento & purificação , Humanos , Tipagem Molecular , Filogenia , Infecções por Roseolovirus/líquido cefalorraquidiano , Infecções por Roseolovirus/genética , Infecções por Roseolovirus/patologia , Transplante Homólogo , Ativação Viral
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