Acute liver dysfunction not resulting from hepatitis virus in immunocompetent children.

Lymphotropic Herpesviruses in Allogeneic Bone Marrow Transplantation

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.

Validation of Roche LightCycler Epstein-Barr Virus Quantification Reagents in a Clinical Laboratory Setting

To identify the role of human herpesvirus 6 (HHV-6), HHV-7, Epstein-Barr virus (EBV) and cytomegalovirus (CMV) in the pathogenesis of pityriasis rosea (PR). Polymerase chain reaction with specific primers for HHV-6 and HHV-7 DNA sequences was performed on the blood and tissue samples of 25 patients with PR and on the blood samples of age- and sex-matched healthy controls. HHV-6, EBV, CMV immunoglobulin M (IgM) and IgG were analysed by enzyme-linked immunosorbent assay, HHV-7 IgM and IgG were analysed by indirect immunofluorescence on the serum samples of the study population. In the patient group, the values were studied 2 weeks later again (second control). There were no differences between the first and second controls of the patients and healthy subjects regarding HHV-6 IgM, HHV-7 IgM, CMV IgM, EBV IgM results. There were significant differences between the first [HHV-6 DNA (2 of 25), HHV-7 DNA (6 of 25)] and second control [HHV-6 DNA (1 of 25), HHV-7 DNA (11 of 25)] of the patients for the blood samples in favour of HHV-7. PR patients showed higher amounts of HHV-6 and HHV-7 DNA positivity when compared with that of healthy subjects. HHV-7 seemed to be more important regarding tissue samples [HHV-6 DNA (7 of 25), HHV-7 DNA (12 of 25) first control, HHV-6 DNA (6 of 25), HHV-7 DNA (12 of 25) second control] as well as blood samples. Though our results failed to support a causal relationship among EBV, CMV and PR, they indicated a possible role for HHV-6 and especially HHV-7 in a group of Turkish patients but other aetiological factors may exist.

Recently, it has been recognized that drug-induced hypersensitivity syndrome (DIHS) is associated with reactivation of human herpesvirus-6 (HHV-6), Epstein–Barr virus (EBV) and cytomegalovirus (CMV). However, whether those viruses have a role in the development of cutaneous drug reactions (CDRs) other than DIHS is not known. To investigate the role of HHV-6, EBV and CMV infections in the etiopathogenesis of different types of CDRs. Eighteen patients with diagnosis of CDR according to the clinical and histopathological findings were evaluated. Real-time polymerase chain reaction (PCR) was used for the detection of EBV, CMV, and HHV-6 DNA in lesional skin biopsy specimens; EBV and CMV DNA in serum samples; and HHV-6 DNA in peripheral blood mononuclear cells. The genome of HHV-6 was detected only in the lesional skin of two patients with DIHS. Epstein–Barr virus and CMV DNA in the skin lesions, EBV and CMV genomes in the serum samples, and HHV-6 DNA in the peripheral blood mononuclear cells were negative in all patients. The patient population was small and did not include all types of CDRs. Also, we had only two patients with DIHS. We had not been able to measure the increase in anti-viral IgG titers in serial serum samples. Epstein–Barr virus and CMV infections do not seem to have a role in the etiopathogenesis of CDRs including DIHS. The association between HHV-6 infection and CDRs is likely to be limited to DIHS.

Non-hepatotropic viral hepatitis and its causative pathogens: The ongoing need for monitoring in children with severe acute hepatitis of unknown etiology.

Prevention and treatment of epstein–barr virus‐associated lymphoproliferative disorders in recipients of bone marrow and solid organ transplants

Idiopathic nephrotic syndrome (INS) is likely a primary immune disorder, but viruses might also be involved in the mechanisms of the disease. Here, we investigate the link between herpesvirus infection and the first manifestation of INS in children. A prospective, multicentre, and population-based case-control study called NEPHROVIR included 164 patients, aged 6 months to 15 years old, newly diagnosed with INS, and 233 controls matched for gender, age, and period of sample. The analysis was done on 124 patients and 196 controls. Epstein-Barr virus (EBV), cytomegalovirus (CMV), human herpesvirus-6 (HHV-6), and human herpesvirus-7 (HHV-7) DNA prevalence at diagnosis were assessed from whole peripheral blood samples, as well as EBV and CMV viral load and seroprevalence. EBV DNA was significantly more prevalent in cases than in controls (50.8 vs 29.1 %; OR = 2.6; p = 0.0002), with no difference in viral load. A significant difference was also found for CMV (11.3 vs 3.6 %; p = 0.02) and HHV-7 (83 vs 72 %; p = 0.02) DNA prevalence between cases and controls. There were significantly more EBV and CMV recent infections or reactivations based on VCA-IgM and CMV IgM in cases than controls, while there were no differences in IgG seroprevalence. The prevalence of positive EBV DNA detection and recent infection or reactivation is higher in children at onset of INS compared to a population matched for age, gender, and time of sampling.

We developed a multiplex real-time PCR assay using 6-carboxyfluorescein, 6-carboxy-4',5'-dichloro-2',7'-dimethoxyfluorescein, and carbocyanine 5-labeled probes to simultaneously quantify Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 (HHV-6) DNA. When previously tested and stored DNA samples were examined, results of the multiplex real-time PCR assay were as sensitive and specific as those of a single real-time PCR assay. The multiplex assay was used to quantify the EBV, CMV, and HHV-6 DNA in 46 transplant recipients. A total of 303 whole-blood and plasma specimens were collected and analyzed. According to the results of the multiplex assay, the detection rates for viral DNA in whole blood and plasma were 23.8% and 5.9% for EBV, 11.2% and 5.3% for CMV, and 12.5% and 2.0% for HHV-6, respectively. All forms of viral DNA were detected more frequently in whole blood than in plasma. During the symptomatic period, EBV DNA was detected in all whole-blood specimens but not in all plasma specimens. Furthermore, the EBV DNA load in whole blood was higher during the symptomatic period than during the asymptomatic period, whereas the EBV DNA load in plasma was similar for both periods. These results demonstrate that whole blood is more suitable for the quantification of EBV DNA in transplant patients. However, a cutoff value with clinical relevance still needs to be determined.

Oral ulcers are common in AIDS patients, with a wide spectrum of underlying causes, including different viruses. In the present study, the presence of cytomegalovirus (CMV), Epstein-Barr virus (EBV) and human herpesvirus-8 (HHV-8) DNA was analysed in 21 biopsies from oral ulcers of 17 male homosexual AIDS patients. The methods used were in situ hybridization (ISH) and the polymerase chain reaction (PCR) with subsequent non-radioactive Southern blot hybridization to confirm the specificity of PCR products. With ISH, 4 biopsies were CMV DNA-positive and 11 contained EBV-DNA. Using PCR, an additional 4 CMV- and 7 EBV-positive samples were detected, and HHV-8 DNA was present in three oral ulcers. Six of the patients (35%) had oral ulcers co-infected by two or three viruses. The overall figures for patients with the detectable EBV-, CMV-, and HHV-8 DNA were 82% (14/17), 35% (6/17) and 18% (3/17), respectively. This is the first study to show the frequent presence of EBV-DNA in oral ulcers of AIDS patients. Because ISH-positivity signifies active virus replication, these results implicate an etiological role of EBV in AIDS-associated oral ulcers. The causal role of HHV-8 has to be considered as well, because this virus was detected in three such ulcers, which were not associated with Kaposi's sarcoma. To conclude, three common members of the herpesvirus family (CMV, EBV, HHV-8) were detected in all but three ulcers in AIDS patients, warranting the inclusion of these viral analyses in the diagnosis of ulcerative lesions of the oral mucosa in all immunosuppressed individuals.

Association of drug-induced hypersensitivity syndrome with viral infection is debated. Human herpesvirus 6 (HHV-6) reactivation has been the most frequently reported infection associated with this syndrome. However, a case of cytomegalovirus (CMV) infection was recently described associated with anticonvulsant-induced hypersensitivity syndrome. We report a case of severe allopurinol-induced hypersensitivity syndrome with pancreatitis associated with Epstein-Barr virus (EBV) infection. Active EBV infection was demonstrated in two consecutive serum samples by the presence of anti-EBV early antigen (EA) IgM antibodies and an increase in anti-EBV EA IgG antibodies, whereas no anti-EBV nuclear antigen IgG antibodies were detected. EBV DNA was detected by polymerase chain reaction (PCR) in peripheral blood mononuclear cells. Reactivation of HHV-6 was suggested only by the presence of anti-HHV-6 IgM antibodies, but HHV-6 DNA was not detected by PCR in the serum. Other viral investigations showed previous infection (CMV, rubella, measles, parvovirus B19), immunization after vaccination (hepatitis B virus), or absence of previous infection (hepatitis C virus, human immunodeficiency virus). We suggest that EBV infection may participate in some cases, as do the other herpesviruses HHV-6 or CMV, in the development of drug-induced hypersensitivity syndrome.

Acute <scp>Epstein‐Barr</scp> virus associated haemophagocytosis in an Asian female: What is the diagnosis?

The role of viral infection in bronchial asthma (BA) is well-known being reflected particularly in GINA. An effect of pneumotropic intracellular persistent herpesviruses on the course of BA is of particular interest. The most common viruses of this group are cytomegalovirus (CMV), EpsteinBarr virus (EBV), and human herpesvirus 6 (HHV-6). The CMV role has been discussed in our previous publications allowing us to focus here on EBV and HHV-6. We examined 167 children with BA that was diagnosed and clinically assessed in accordance with the current national clinical guidelines. Patients with controlled asthma (70 patients), partially controlled and uncontrolled asthma (97 patients) were stratified into several groups. The detection of EBV and HHV-6 DNA was carried out in throat swabs by PCR; the level of total and virus-specific IgA, IgM, IgG, IgE as well as serum level of IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-8 and TNF was assessed by enzyme linked immunosorbent assay; lymphocyte subpopulations were analyzed by flow cytometry. The dose of topic GCS was taken into account with reference to the fluticosone equivalent; the function of external respiration was studied by spirometry. It was revealed that EBV DNA was found in 14.2% of cases, whereas HHV-6 in 19.0% of cases, but 14.4% and 52.4% of patients, respectively, shed no pathogen-linked DNA. At the same time, patients with uncontrolled BA are significantly more likely to shed the pathogen DNA, so that EBV is released in 17.1% vs 4.5% of patients with controlled BA, HHV-6 in 19.5% vs 13.6%. On the contrary, children with controlled BA were significantly more often (63.6% vs 51.2%) negative for viral DNA shedding. Moreover, virus shedding was paralleled with higher levels of IL-5: it was as high as 0.91 pg/ml and 0.29 pg/ml for EBV and HHV-6, respectively; those shedding DNA of both pathogens vs no shedding had IL-5 at level of 0.25 pg/ml vs 0.11 pg/ml. Similar pattern was observed for higher total IgE: 184.5 IU for EBV, 113.1 IU for HHV-6, and 371.7 IU shedding both viruses vs 95.2 IU in without DNA pathogen shedding; lower level of FEV1: EBV 96.6%, HHV-6 98.8%, and 106.2% in patients shedding both viruses vs 109.8% in patients not shedding the pathogen DNA. Patients shedding EBV DNA require higher doses of topic GCS to achieve disease control: EBV 325.0 mg, HHV-6 186.4 mg; shedding both viruses 328.1 mg of topic GCS vs 198.6 mg in patients without pathogen DNA shedding. Thus, activation of both EBV and HHV-6 worsens BA control and aggravates its course, but EBV persistence has a more pronounced effect on the course and control of the disease.

Epstein–Barr Virus Infection Induces Aberrant TLR Activation Pathway and Fibroblast–Myofibroblast Conversion in Scleroderma

Objective To explore the clinical features of cytomegalovirus (CMV) and Epstein-Barr virus (EBV) infection after second hematopoietic stem cell transplantation (HSCT). Methods Twenty-five patients after second HSCT from Sep. 2009 to Oct. 2016 were collected, and CMV and EBV DNA in peripheral blood was detected regularly by polymerase chain reaction (PCR). Factors associated were compared by univariate analysis. Results The total incidence of CMV infection was 52.0% (13/25) after second HSCT. The incidence of CMV infection was 100% (2/2), 33.3% (5/15) and 75% (6/8) in bone marrow group, peripheral blood stem cell group, and mixed group, respectively. Stem cell sources were significantly correlated with CMV infection (P=0.038), however, there was no significant difference in CMV infection rate among three groups (P>0.05). None of preconditioning regimen, GVHD prophylaxis programs or severity of aGVHD were correlated with CMV infection after second HSCT (P>0.05). The total incidence of EBV infection was 24.0% (6/25) after second HSCT. The incidence of EBV infection was 100% (2/2), 6.7% (1/15) and 37.5% (3/8) in bone marrow group, peripheral blood stem cell group, and mixed group, respectively. Stem cell sources were significantly correlated with EBV infection (P=0.008). The EBV infection rate in bone marrow group was significantly higher than that in peripheral blood group (P=0.022), however, no significant differences were found between bone marrow group and mixed group, as well as between peripheral blood group and mixed group (P>0.05). Transplant methods were significantly correlated with EBV infection (P=0.007). The EBV infection rate in haplo-identical HSCT group (71.4%) was significantly higher than that in HLA-matched sibling HSCT group (0%) and autologous HSCT group (0%) (P=0.021 and 0.028), however, no significant differences were found between any other two groups (P>0.05). None of preconditioning regimen, GVHD prophylaxis programs or severity of aGVHD were correlated with EBV infection after second HSCT (P>0.05). Conclusion The incidence of CMV and EBV infection in patients undergoing second HSCT is high. Stem cell sources and transplant methods are associated with CMV and EBV infection after second HSCT. Key words: Hematopoietic stem cell transplantation; Cytomegalovirus; Epstein-Barr virus