Detection of herpesvirus DNA in the large intestine of patients with ulcerative colitis and Crohn's disease using the nested polymerase chain reaction.

The prevalence of herpesvirus DNA was examined in inflammatory bowel disease tissue. DNA was extracted from resection and biopsy specimens of the large intestine from patients with ulcerative colitis (n = 21), patients with Crohn's disease (n = 29), and patients with noninflammatory bowel disease (controls) (n = 21). The nested polymerase chain reaction was used to detect viral DNA using primer pairs specific for either cytomegalovirus (CMV), herpes simplex virus 1 (HSV1), human herpesvirus 6 (HHV6), varicella zoster virus (VZV), or Epstein Barr virus (EBV). HSV1 and VZV DNA were not detected in any of tissue samples. There was a high prevalence of CMV (81%), HHV6 (76%), and EBV (76%) DNA in ulcerative colitis tissue compared to Crohn's disease tissues (CMV 66%, HHV6 45%, EBV 55%). Control tissue had a relatively low frequency of CMV (29%) and EBV (19%) DNA but a prevalence of HHV6 DNA similar to that of ulcerative colitis (86%). However, the simultaneous presence of HHV6 and CMV and/or EBV DNA in ulcerative colitis tissue (76%) was much greater than in either Crohn's disease tissues (38%) or control tissue (29%) (P < 0.05). There was a low prevalence of CMV, HHV6, and EBV DNA in peripheral blood mononuclear cells from all patient groups. CMV and EBV are capable of reactivating HHV6: the high prevalence of coexistent HHV6 infection with either or both of these two viruses in ulcerative colitis tissue suggests that they may play a synergistic role in the pathogenesis of this disease.     

Avidity of IgG antibodies to human herpesvirus-6 distinguishes primary from recurrent infection in organ transplant recipients and excludes cross-reactivity with other herpesviruses

Saliva and peripheral blood mononuclear cells from three patients, two with lymphoproliferative disorders and one suffering from multiple sclerosis, were examined for the presence of human herpesvirus-6 (HHV-6) genome by using the polymerase chain reaction and Southern blot analysis. The search for anti-HHV-6 antibodies, carried out in the sera of the same cases by an immunofluorescence assay, was negative in two cases at the lowest dilution used (1:40). These three patients had a high number of HHV-6 specific sequences in uncultured peripheral blood mononuclear cells, which are thought to be a normal site of viral latency although, in healthy individuals, the infected cells are extremely rare. In order to gain some insight into the state of the viral genome in this latent HHV-6 infection, we used pulsed field gel electrophoresis to separate HHV-6 DNA directly from HHV-6 (strain GS) infected HSB-2 cells and from the peripheral blood mononuclear cells of these three patients. Our study showed the presence of intact viral genome, of the expected length of 170 kb, persisting as free extrachromosomal element in the HSB-2 cells but not in patients' peripheral blood mononuclear cells. On the other hand, in strong contrast with the results obtained in infected HSB-2 DNA, the restriction analysis of the three patients' peripheral blood mononuclear cell DNA showed fragments of molecular weight constantly higher than the 170 kb segment, indicating that the viral sequences are linked to high molecular weight cellular DNA. Our findings are consistent only with a latent infection in which HHV-6 is integrated in vivo and suggest that pulsed field gel electrophoresis analysis is well worth using to evaluate the presence of integrated, intact, or fragmented viral genomes in HHV-6 associated lymphoproliferative diseases and immune disorders. © 1993 Wiley-Liss, Inc.     

A serological investigation of human herpesvirus 6 infections in liver transplant recipients and the detection of cross-reacting antibodies to cytomegalovirus.

Sera from 50 orthotopic liver transplant recipients were examined for antibodies to human herpesvirus 6 (HHV-6) and cytomegalovirus (CMV), and the findings correlated with the clinical condition of the patients. Both primary and secondary HHV-6 infections were detected serologically following liver transplantation. Interpretation of serological assays is complicated by CMV and HHV-6 antibody cross reactions which were common. Sera from 5 patients became HHV-6 antibody negative following absorption with CMV infected cells. Thirty patients were initially seronegative for HHV-6 antibodies, 12 remained so following transplantation, 5 developed cross reacting antibodies, and 13 seroconverted. The seroconversions occurred at 4 to 8 weeks post-transplant in the same time period as CMV antibody rises. HHV-6 IgM was detected in only 4 of the 13. Of the 7 patients who had serological evidence of active HHV-6 infections but no evidence of CMV infection, 4 (56%) had fever, 1 (14%) hepatitis, 1 (14%) lung dysfunction, and 3 (42%) neurological disorders. In the 12 patients who remained HHV-6 antibody negative, there were fewer fevers and neurological disorders.     

Confirmation of the low clinical effect of human herpesvirus-6 and -7 infections after renal transplantation

Human herpesvirus‐6 and ‐7 (HHV‐6 and HHV‐7) may lead to pathological manifestations in renal transplant recipients. The aim of this study was to investigate beta‐herpesvirus infections in 50 adult kidney transplant recipients after transplantation to examine the effect, interactions, and pathogenic consequences of infection and the effect of immunosuppressive regimens and Human cytomegalovirus (HCMV) prophylaxis with VACV. Beta‐herpesviruses loads in the blood of 50 adult kidney transplant recipients over a 6‐month period after transplantation and 198 blood donors were determined using polymerase chain reaction. The rate of HHV‐6 detection in peripheral mononuclear cells (PBMCs) was higher in patients with end‐stage renal disease and during the post‐transplantation follow‐up than in healthy subjects (33% and 68% vs. 12%, respectively). The detection rate of HHV‐7 in PBMCs was similar between patients, both before grafting and during the follow‐up for transplant recipients (69% and 88%, respectively), and healthy subjects (78%), and correlated with the number of lymphocytes. HCMV in plasma was detected only in patients during the post‐transplant period (24%). VACV prophylaxis had no negative effect on the replication of HHV‐6 or HHV‐7, and univariate analyses demonstrated associations between HHV‐6 infection and acute graft rejection [Odds ratio (OR) = 2.94, 95% confidence interval (CI), 1.05–8.2, P = 0.04], and between HHV‐7 infection and cholestasis [OR = 2.61 (95% CI, 1.08–6.3), P = 0.03]. Immunosuppressive regimens had no effect on beta‐herpesviruses infections. This study revealed the differing behavior of HCMV, HHV‐6, and HHV‐7 in kidney transplant recipients, and confirmed the association of HHV‐6 with graft rejection. J. Med. Virol. 84:450–456, 2012. © 2011 Wiley Periodicals, Inc.     

Epstein-Barr virus in angioimmunoblastic T-cell lymphomas

Epstein-Barr virus (EBV) has been proposed as a possible infective agent involved in the pathogenesis of angioimmunoblastic lymphadenopathy (AIL), a progressive and often fatal lymphoproliferative disorder. We have studied 19 cases of AIL-like lymphomas for the presence of EBV using a sensitive in situ hybridization technique based on the detection of Epstein-Barr encoded RNAs with digoxigenin-labelled oligonucleotide probes. EBV was found in 11 cases; in seven of these EBV was detected in occasional cells. Immunocytochemical studies to investigate viral gene expression, revealed the presence of EBV-encoded latent membrane protein only in those cases which had appreciable numbers of positive cells by in situ hybridization. The intensity of staining varied from case to case and the overall proportion of cells staining for latent membrane protein in a given case was considerably less than that by in situ hybridization. In situ hybridization for cytomegalovirus and human herpes virus type-6 was negative in all cases. We discuss these findings in the light of the proposed role of EBV in the pathogenesis of AIL and conclude that the presence of EBV is a consequence of the disease rather than the cause.     

Hepatitis C virus seroprevalence in Italian haemophiliacs injected with virus-inactivated concentrates: five year follow-up and correlation with antibodies to other viruses.

The overall prevalence of anti-HCV antibody in a group of 125 haemophiliacs was 62%. Four patients who had never received replacement therapy were anti-HCV negative. Of the 121 patients injected regularly with commercial concentrates, 76 were already anti-HCV seropositive in 1985 and remained so throughout the follow-up. Two patients seroconverted in 1987 without obvious signs or symptoms of hepatitis. Our patients were treated with dry heat-treated concentrates since 1985 and with wet heat- or solvent/detergent-treated concentrates since 1988. The absence of further seroconversions and of symptoms of acute post-transfusion non-A, non-B hepatitis since 1988 suggest that present virucidal treatments of concentrates are effective in preventing HCV transmission. Anti-HCV positivity appeared to be unrelated to the type and degree of haemophilia as well as to the presence of antibodies to hepatitis B virus, human immunodeficiency virus type 1, and human herpesvirus type 6.     

Risk factors and clinical consequences of human herpesvirus 7 infection in paediatric haematopoietic stem cell transplant recipients

Human herpesvirus 7 (HHV-7) is the least studied beta-herpesvirus in transplant settings. This prospective study examined the activity of HHV-7 during the first 12 weeks post-stem cell transplant in 59 paediatric patients. The presence of HHV-7, human cytomegalovirus (HCMV) and human herpesvirus 6 (HHV-6) in blood was monitored weekly by a multiplex nested polymerase chain reaction. Overall, 33 (55.9%) patients had one or more surveillance blood sample(s) positive for HHV-7. In contrast to HCMV and HHV-6, no obvious peak time of reactivation was observed for HHV-7. The occurrence of HHV-7 DNAaemia showed a significant negative association with HHV-6 (P=0.022), but with no association with HCMV. A significant higher positive rate for HHV-7 was found in autologous versus allogeneic (P=0.002), and in peripheral blood versus umbilical cord/marrow (P<0.001) transplant. Acyclovir had no effect, whereas ganciclovir was associated with a lower rate of HHV-7 reactivation (P=0.009). One patient died of HHV-7 associated brain stem encephalitis. The administration of colony stimulating factor, occurrence of acute graft versus host disease, time to neutrophil and platelet engraftment showed no significant association with the occurrence of HHV-7 DNAaemia.     

Sensitivity of human herpesvirus 6 and other human herpesviruses to the broad-spectrum antiinfective drug artesunate

BackgroundAntiviral therapy for HHV-6 infection with conventional anti-herpesviral drugs is problematic so novel drugs are required. Artesunate is a well-tolerated drug approved for malaria therapy which possesses antiviral activity.ObjectiveThe artesunate sensitivity of HHV-6 was analyzed and compared to that of several other human herpesviruses.Study designCultured human cells were productively infected with strains of HHV-6 or other human herpesviruses to measure artesunate inhibition of viral protein synthesis (Western blot analysis) or viral genome replication (qPCR), and to determine IC₅₀ values by immunofluorescence or plaque reduction assays.ResultsSensitivity of HHV-6 to artesunate was demonstrated with an IC₅₀ of 3.80 ± 1.06 μM. This is in a range similar to IC₅₀ values for HCMV and EBV. Artesunate treatment of HHV-6-infected cells significantly reduced viral early and late protein synthesis that occurred in the absence of drug-induced apoptosis or necrotic cytotoxicity. HHV-6A genome replication was markedly reduced by artesunate.ConclusionsArtesunate possesses anti-HHV-6 activity in vitro and may be useful for treatment of HHV-6 infections.     

Case report: human herpesvirus 6 reactivation associated with colitis in a lung transplant recipient

Whereas human herpesvirus 6 (HHV-6) reactivation is frequent in solid organ transplant recipients, symptomatic disease is rare. A case of colitis associated with HHV-6B reactivation was observed in a lung transplant recipient. This case report suggests that symptomatic HHV-6 infection may occur in the absence of detectable viremia.     

Co-existence of Herpes simplex virus type 2 and two other oncoviruses is associated with cervical lesions in women living with HIV in South-Western Nigeria

BackgroundThe prevalence of Herpes simplex virus type 2 (HSV-2) in cervical lesions is under-reported, especially in Human immunodeficiency virus (HIV), Epstein-Barr virus (EBV) and Human Papillomavirus (HPV) infected persons.ObjectivesThis study determined the prevalence of viral mono-infections, co-infections and squamous cell intraepithelial lesions (SIL) in HIV seropositive (HIV+) and HIV seronegative (HIV-) women.MethodsThis study included HIV+ and HIV- women (105 each). Cervical smears and viral antibodies were evaluated by Papanicolaou''s technique and ELISA method, respectively.ResultsThe prevalence of HSV-2, HPV and EBV infections, and SIL were higher in HIV+ women (75.2, 41.9, 41 and 32.4%) than in HIV- women (45.7, 26.7, 26.7 and 13.3%) at p< 0.0001, p= 0.029, 0.041 and 0.002, respectively. Higher prevalence of viral mono-infection and tri-infection was observed in HIV+ women (43.8 and 24.8%) than in HIV- women (27.6 and 8.6%) at p= 0.021, and 0.003, respectively. The prevalence of SIL was also higher in HIV+ women with viral mono-infection, bi-infection and tri-infection (15.2, 42.9, and 53.8%) than in HIV- women (6.9, 12.5, and 44.4%) at p= 0.468, 0.041, and 0.711, respectively.ConclusionThis study suggests that the high prevalence of SIL in HIV+ women could be associated with viral co-infections.     

Viral reactivation in the lungs of patients with severe pneumonia is associated with increased mortality,a multicenter,retrospective study

Viral reactivation is widespread in patients with severe pneumonia, yet the landscape of viral reactivation in the lungs is not well-known. This study aims to assess the landscape and clinical features of viral reactivation in the early onset of severe pneumonia in ICU patients. The clinical data from 97 patients were collected retrospectively from the intensive care units of five teaching hospitals between June 2018 and July 2021. Metagenomic next-generation sequencing (mNGS) of the bronchoalveolar lavage fluid (BALF) was performed at the onset of severe pneumonia. Cytomegalovirus (CMV), herpes simplex virus-1 (HSV-1), and Epstein-Barr virus (EBV) were the most common reactivated viruses in the lower respiratory tract of patients with severe pneumonia. After adjusting for the risk of confounding and competition of age, sex, sequential organ failure assessment, acute physiology chronic health assessment II and immunosuppression status, viral reactivation resulted in an overall 2.052-fold increase in 28-day all-cause mortality (95% CI: 1.004–4.194). This study showed that CMV, HSV-1, and EBV were the most common reactivated viruses in the lungs of patients with severe pneumonia. The existence of viral reactivations was associated with an increased risk of mortality. The simultaneous reactivation of multiple viruses needs to be considered in the design of clinical trials.     

Productive herpes simplex virus in brain of elderly normal subjects and Alzheimer's disease patients

It was previously shown that herpes simplex virus type 1 (HSV1) DNA resides latently in a high proportion of aged brains and that in carriers of the type 4 allele of the apolipoprotein E gene (APOE-epsilon4), it confers a strong risk of Alzheimer's disease. It was suggested that initial entry of brain by HSV1 and any subsequent reactivation(s) would cause a type of limited encephalitis, the resulting damage being more harmful in APOE-epsilon4 carriers. Reactivation(s) would induce synthesis of intrathecal antibodies; these are long-lived after herpes simplex encephalitis so they were sought in cerebrospinal fluid (CSF) of Alzheimer's disease patients and age-matched normal subjects. Intrathecal antibodies to human herpesvirus 6 (HHV6) were also sought as DNA of this virus has been detected previously in a high proportion of Alzheimer's disease brains. Antibody indices for HSV and HHV6 were measured using indirect ELISA for IgG antibody, and single radial immunodiffusion was used for albumin, in serum and CSF. A raised antibody index (>1.5) indicative of virus-specific intrathecal HSV1 IgG synthesis was found in 14/27 (52%) Alzheimer's disease patients and 9/13 (69%) age-matched normals (difference non-significant). A raised antibody index to HHV6 was detected in 22% of the Alzheimer's disease patients and in no normals, so presumably this virus either did not reactivate in brain or it elicited only short-lived intrathecal antibodies. The HSV1 results confirm the original PCR findings that show the presence of HSV1 DNA sequences in many elderly brains, and indicate also that the whole functional HSV1 genome is present, and that the virus has replicated.     

Malignant lymphoma induction of rabbits with oral spray of Epstein-Barr virus-related herpesvirus from Si-IIA cells (HTLV-II-transformed Cynomolgus cell line): A possible animal model for Epstein-Barr virus infection and subsequent virus-related tumors in humans

Malignant lymphoma (ML) was Induced in eight of nine rabbb inoculated by oral spray of the cell-free pellets from SI-IIA culture (MLV-ll-transformed leukocyte cell line of the Cynomolgus-producing Epsteln-Barr virus (EBV)-related herpesvirus) after 64–141 days. None of the rabbits inoculated with EBV from B-95–8 cells or HTLV-II from MOT cells developed ML. Malignant lymphomas were usually of diffuse, large-cell or mixed type. HTLV-II infection was excluded by the polymerase chain reaction (PCR) and the particle agglutination test. EBV-encoded RNA-1 and EBV-related DNA were detected in the tumor tissues by In situ hybridization and PCR, respectively. Anti-viral capsid antigen of EBV antibody (anti-VCA) was observed 3 weeks after oral inoculation of Si-IIA cell-free pellets. Polymerase chain reaction revealed continuous detection of EBV-related virus DNA in the peripheral blood leukocytes from 3 days after oral inoculation. These results show that ML induced orally wtth SI-IIA cell-free pellets was caused by EBV-related herpesvirus harbored by Si-IIA cells. Oral spray of EBV from B-95–8 also induced EBV Infection in rabbits, which was confirmed both by the presence of anti-VCA and by PCR. These oral infection and mallgnant lymphoma induction systems of rabbit using EBV-related virus from Si-IIA or human EBV are useful animal models for the study of EBV infection and EBV-related lymphomas in humans.     

Search for human herpesvirus 6 and human cytomegalovirus in bronchoalveolar lavage from patients with human immunodeficiency virus-1 and respiratory disorders

Virus isolation and viral DNA detection by the polymerase chain reaction were used to investigate the presence of human herpesvirus 6 (HHV-6) and human cytomegalovirus (HCMV) in bronchoalveolar lavage from 34 human immunodeficiency virus-1 (HIV-1)-infected patients with respiratory disorders. The aim was to assess the presence of reactivated HHV-6 in lung tissues for a subsequent evaluation of the frequency of virus involvement in respiratory clinical manifestations in the course of HIV-1 infection. Bronchoalveolar lavage samples were tested for the presence of HCMV, as a routine investigation within a protocol monitoring opportunistic infections in symptomatic HIV-1 patients. Whereas HCMV DNA was detected by the polymerase chain reaction in 12 bronchoalveolar lavage specimens, 10 of which were also positive for virus isolation, all samples were negative for HHV-6 by both virological procedures. The HHV-6 DNA finding in bronchoalveolar lavage from an HIV-1-seronegative patient with renal carcinoma, investigated accidentally together with the bronchoalveolar lavage specimens from HIV-1 seropositive patients, stressed the HHV-6 polymerase chain reaction-negative results in the bronchoalveolar lavage samples under study. It is concluded that the lung may be a target organ for HCMV infection in HIV-1-seropositive patients affected by respiratory symptoms but that this does not seem to be the case for HHV-6. © 1996 Wiley-Liss, Inc.     

Case report: primary human herpesvirus-6 associated with an afebrile seizure in a 3-week-old infant.

We describe a 3-week-old male infant with an afebrile seizure in whom serologic and polymerase chain reaction (PCR) findings support concomitant primary human herpesvirus 6 (HHV-6) infection. Although HHV-6 infection has been associated with first-time febrile seizures and encephalitis in both immunocompetent and immunocompromised hosts, it has not been associated previously with afebrile seizures in healthy infants. This report provides additional evidence of the neuropathogenic potential of HHV-6.     

Infection of infants with human herpesvirus type 6 may be associated with reduced allergic sensitization and T‐helper type 2 development

Background Epidemiological studies point to an inverse relationship between microbial exposure and the prevalence of allergic diseases. The underlying mechanism for this observation remains largely unknown, as well as the nature of the microbes involved. Objective To investigate the effects of early infection with human herpesviruses (HHVs) on IgE formation and T‐helper type 2 (Th2) development in infants. Methods Serum was collected from children aged 18 months and assessed for IgE to common allergens and IgG to five common herpesviruses. Cord blood plasmacytoid dendritic cells (pDC) were exposed to HHV type 6 in vitro and mixed with allogeneic cord blood CD4⁺ T cells. Cytokine levels were determined by ELISA and by flow cytometry. Results We found that children seropositive at 18 months of age to HHV type 6 were significantly less often IgE sensitized than seronegative children [odds ratio (OR): 0.08, 95% confidence interval (CI): 0.009–0.68]. HHV type 6 also decreased the production of the Th2‐associated cytokines IL‐5 and IL‐13 by CD4⁺ T cells when co‐cultured with allogeneic cord blood pDC. This was associated with an increased production of IFN‐α by pDC exposed to HHV type 6. Conclusion These data indicate that an early childhood infection with HHV type 6 could down‐regulate Th2 responses and reduce IgE formation to common allergens in a young child.     

Multiplex PCR-based DNA array for simultaneous detection of three human herpesviruses, EVB, CMV and KSHV

Human lymphotropic herpesviruses, Epstein-Barr virus (EBV), cytomegalovirus (CMV) and Kaposi's sarcoma-associated herpesvirus (KSHV) are responsible for a wide variety of human diseases. Due to an increase in diseased states associated with immunosuppression, more instances of co-morbid infections with these herpesviruses have resulted in viral reactivations that have caused numerous fatalities. Therefore, the development of rapid and accurate method to detect these viruses in immunocompromised patients is vital for immediate treatment with antiviral prophylactic drugs. In this study, we developed a new multiplex PCR method coupled to DNA array hybridization, which can simultaneously detect all three human herpesviruses in one single cell sample. Multiplex PCR primers were designed to amplify specific regions of the EBV (EBER1), CMV (IE) and KSHV (LANA) viral genomes. Pre-clinical application of this method revealed that this approach is capable of detecting as few as 1 copy of the viral genomes for KSHV and CMV and 100 copies of the genome for EBV. Furthermore, this highly sensitive test showed no cross-reactivity among the three viruses and is capable of detecting both KSHV and EBV viral genomes simultaneously in the lymphoblastoid cells that have been double infected with both viruses. Thus, this array-based approach serves as a rapid and reliable diagnostic tool for clinical applications.     

Contribution of herpesviruses in the progression of periodontal and peri‐implant diseases in systemically healthy individuals

The aim of this review was to assess the contribution of herpesviruses in the subgingival oral biofilm in the progression of periodontal and peri‐implant diseases in systemically healthy individuals. The literature review was customized to summarize the pertinent information for the following reasons: (1) A systematic review regarding the role of herpesviruses in the etiopathogenesis of periodontal disease has recently been published; and (2) a limited number of studies have assessed the association of herpesviruses with peri‐implant diseases. To date, five observational studies have assessed the presence of herpesviruses in the subgingival oral biofilm of individuals with peri‐implant diseases. In these studies, dental implants were in place for up to approximately 8 years. In two studies, human cytomegalovirus (HCMV) was more often isolated from the peri‐implant sulci of sites with than without peri‐implantitis. In one study, a low prevalence of HCMV compared with Epstein‐Barr virus (EBV) was associated with the progression of peri‐implantitis. In previous studies, the presence of EBV in the subgingival oral biofilm was associated with the onset of peri‐implantitis and peri‐implant mucositis, respectively. Major limitations of the studies assessed were the absence of blinding and lack of power analysis for sample size estimation. In conclusion, the presence of herpesviruses in the periodontal and peri‐implant subgingival oral biofilm is an indicator of periodontal and peri‐implant diseases in systemically healthy individuals; however, further studies with a statistically justified sample‐size are needed to understand and refine this association.