Chronic active Epstein-Barr virus infection (CAEBV) successfully treated with allogeneic peripheral blood stem cell transplantation.

We report a pediatric case of CAEBV and T cell-based Hodgkin's-like disease successfully treated with allo PBSCT from an HLA-matched sibling. The diagnosis of CAEBV was made from clinical signs and the presence of the EBV genome in PBMC and tumor cells. Conditioning with busulfan (BU) + etoposide (VP16) + cyclophosphamide (CY) was effective and well tolerated. EBV was totally eradicated by 3 months after allo PBSCT. Although she suffered from chronic GVHD of the liver, she has been well and free of disease for 47 months since PBSCT. We suggest allo PBSCT for CAEBV as a potent therapeutic strategy for eradication of the EBV genome and allowing immunological reconstitution.

Fatal hemophagocytic syndrome after living-related liver transplantation: a report of two cases.

BACKGROUND: Hemophagocytic syndrome (HPS) is a serious hematological disorder caused by activated T lymphocytes in immunologically compromised patients. There is no report of HPS in liver transplant recipients. METHODS: Among 135 patients who underwent living-related liver transplantation between June 1990 and October 2000, HPS developed in two pediatric patients (1.5%) on the 15th and 134th postoperative day, respectively. The courses of these patients were evaluated. RESULTS: The cause of HPS was unknown in patient 1 and suspected to be Epstein-Barr virus infection in patient 2. The course of patient 2 was also complicated by posttransplant lymphoproliferative disorder. Both patients had high fever, pancytopenia, coagulopathy, and marked elevation of serum-soluble interleukin 2 receptor, serum ferritin, and urine beta2-microglobulin levels. The diagnosis was established based on clinical findings, laboratory data, and bone marrow biopsy. Both patients died in an acute course despite intensive care. CONCLUSIONS: HPS should be recognized as a severe hematological complication in liver transplant patients. Prompt institution of adequate treatment is necessary to prevent fatality.

Serum extracellular superoxide dismutase in pediatric patients with various diseases as judged by an ELISA.

Extracellular superoxide dismutase (EC-SOD) concentration was measured in sera from 141 patients with 20 forms of infantile diseases including IDDM, SLE and epilepsy, 31 healthy children (controls), and 21 healthy young men by an enzyme-linked immunosorbent assay using a polyclonal antibody against human lung EC-SOD. Serum from patients with IDDM and fever of unknown origin had a significantly (p<0.05) lower concentration of EC-SOD than control serum. Part of sera from patients with the seven forms of diseases (SLE, viral infections, epilepsy, nephrosis, hyperthyroidism, hepatic disease, and Reye syndrome), on the other hand, had a greatly high concentration of EC-SOD, albeit not statistically significant. This SOD isoenzyme profile appears to be specific to each pediatric disease.

Development of EBV-positive T-cell lymphoma following infection of peripheral blood T cells with EBV.

Chronic active Epstein-Barr virus (EBV) infection is manifested clinically by the persistence of infectious mononucleosis-like symptoms or its complications for a prolonged period ranging from one to several years. This syndrome may include severe disease manifestations and can be fatal. The role of EBV in the pathogenesis of chronic active EBV infection has been unclear. We investigated two Japanese patients with severe chronic active EBV infection who subsequently developed EBV-positive T-cell lymphoma. We found that the patients had evidence of EBV infection in the peripheral blood CD4+ T-cells 19 and 3 months, respectively, before the T-cell lymphoma was diagnosed. The lymphomas were infected with monoclonal EBV and expressed the EBV latency genes EBNA-1, LMP-1, and LMP-2A, a virus latency pattern referred to as latency II. Genetic studies showed that the virus detected in the T-cell lymphoma was indistinguishable from the virus in the peripheral blood CD4+ T-cells. These studies support an important pathogenetic role of T-cell infection with EBV in chronic active EBV infection and in the EBV-positive T-cell lymphoma that followed.

A syndrome of peripheral blood T-cell infection with Epstein-Barr virus (EBV) followed by EBV-positive T-cell lymphoma.

The role of Epstein-Barr virus (EBV) in the pathogenesis of severe, chronic active EBV infection and its complications is unclear. We investigated two Japanese patients diagnosed with severe, chronic active EBV infection who subsequently developed EBV-positive T-cell lymphoma. The patients displayed abnormally high antibody titers to EBV antigens, and had evidence of peripheral blood CD4(+) T-cell infection with EBV, 19 months and 3 months, respectively, before the diagnosis of EBV-positive T-cell lymphoma. The lymphomas were infected with monoclonal EBV and expressed the EBV latency genes EBNA-1, LMP-1, and LMP-2. Genetic studies showed that the virus detected in the T-cell lymphoma was indistinguishable, with respect to type and previously defined LMP-1 and EBNA-1 gene variations, from virus detected in the peripheral blood T cells 19 months earlier. These studies support an important pathogenetic role of T-cell infection with EBV in chronic active EBV infection and in the EBV-positive T-cell lymphoma that followed.

Efficacy of quantitative analysis of Epstein-Barr virus-infected peripheral blood lymphocytes by in situ hybridization of EBER1 after living-related liver transplantation: a case report.

BACKGROUND: We describe a 1-year-old female who underwent living-related liver transplantation for biliary atresia and developed Epstein-Barr virus (EBV)-related posttransplant lymphoproliferative disorder. This disorder was resolved after withdrawal of immunosuppression therapy and administration of a high dose of acyclovir. METHODS: To quantify the extent of EBV activation and EBV load in peripheral blood, we measured the levels of EBV-infected peripheral lymphocytes by in situ hybridization (ISH) of EBV-encoded small mRNA 1 (EBER1). RESULTS: The decline in the number of EBER1-positive lymphocytes (from 362/50,000 mononuclear cells to 0/50,000) after treatment was in accord with the patient's clinical improvement. CONCLUSIONS: This finding showed that quantitative analysis of EBV-infected peripheral lymphocytes by ISH of EBER1 is very useful for monitoring the EBV load and response to treatment of patients with EBV-related disorders. Furthermore, ISH may become an important tool for the early diagnosis and prevention of life-threatening posttransplant lymphoproliferative disorder in posttransplant patients.

[A quantitative analysis of the cells infected with Epstein-Barr virus in the peripheral blood mononuclear cells derived from the patients with infectious mononucleosis].

The quantitative analysis of the cells infected with Epstein-Barr virus was performed on the peripheral blood mononuclear cells from the patients with infectious mononucleosis, by using in situ hybridization with Epstein-Barr virus encoded small nuclear RNA1 (EBER1). An alkaline-phosphatase conjugated oligonucleotide probe complementary to EBER1 was used as an antisense probe, while oligonucleotide DNA probe compatible with the sequence of EBER1 was used as a sense probe, control probe. The EBER1 positive cells on the slide-glass were enumerated microscopically. In situ hybridization revealed that 50,000 peripheral blood mononuclear cells from the patients in the acute phase of infectious mononucleosis contained 35 +/- 36 cells infected with Epstein-Barr virus (n = 11). The cells infected with Epstein-Barr virus apparently decreased in the convalescence of all the patients with infectious mononucleosis and the mean of the cells infected with Epstein-Barr virus was 3 +/- 4 in the convalescence (n = 6) (p < 0.02). On the other hand, no positive cells were detected in healthy individuals with past-infection of Epstein-Barr virus (n = 10) or without any previous Epstein-Barr virus infection (n = 11). The striking increase of the cells with Epstein-Barr virus genome was clearly demonstrated in the peripheral blood mononuclear cells from the patients with infectious mononucleosis.

Fatal Epstein-Barr virus-associated hemophagocytic syndrome.

A virus-associated hemophagocytic syndrome is characterized by high fever, liver dysfunction, coagulation abnormalities, pancytopenia, and a benign histiocytic proliferation with prominent hemophagocytosis in bone marrow, lymph node, spleen, and liver. We describe six Japanese children with fatal Epstein-Barr virus (EBV)-associated hemophagocytic syndrome. Five of the six patients had serologic evidence of primary EBV infection at the onset of their diseases. EBV genomes were detected in all the patients by Southern blot hybridization or the polymerase chain reaction. Furthermore, clonality analysis of the EBV genome showed that EBV-infected cells proliferated monoclonally or biclonally in three examined patients. In situ hybridization study using EBV-encoded RNA 1 (EBER1) showed that EBER1 was detected in one of two examined liver tissues, which localized in hepatocytes.

Augmented production of interleukin-8 in cerebrospinal fluid in bacterial meningitis.

Interleukin-8 (IL-8) elaborated by monocytes and endothelial cells is a cytokine which is responsible for adhesion of leucocytes to vascular endothelium and migration of neutrophils into the cerebrospinal fluid (CSF) from the intravascular space. The inflammation in meningitis is elicited by the cytokine release from leucocytes which encounter micro-organisms in the arachnoid or subarachnoid space. In bacterial meningitis, tumour necrosis factor (TNF), IL-1 and IL-6 are produced vigorously, and initiate and augment the inflammation in the central nervous system. In this study, utilizing a quantitative immunometric sandwich enzyme immunoassay, the concentration of IL-8 was investigated in the CSF of patients with bacterial meningitis, patients with aseptic meningitis, and patients with gastroenteritis who served as controls. The IL-8 concentration was markedly higher in the CSF of patients with bacterial meningitis (224 +/- 2.57 pg/ml; mean +/- SD) than in the CSF of patients with aseptic meningitis (less than 30 pg/ml). The IL-8 level in the CSF of patients with aseptic meningitis did not differ from that in the CSF of the patients with gastroenteritis (less than 30 pg/ml). The augmented production of IL-8 in CSF may account for the inflammation in bacterial meningitis being more severe than that in aseptic meningitis.

[Electrocardiographic study of sinus bradycardia associated with enterohemorrhagic Escherichia coli O157: H7 infection].

A severe outbreak of hemorrhagic colitis occurred at a kindergarten in Saitama, Japan in October, 1990. Children who were affected by enterohemorrhagic E. coli O157: H7 infection showed apparent bradycardia as well as severe bloody diarrhea, generalized convulsion, or hemolytic uremic syndrome. Cardiac involvement such as bradycardia observed in the patients of this outbreak has not been described in previous reports about EHEC infection, while bradycardia has been well known in typhoid fever due to salmonella typhosa infection. Electrocardiographic examination was performed to evaluate bardicardia, utilizing electrocardiography at rest and Holter's twenty-four hour electrocardiography. In the report, we demonstrate that the bradicardia was due to reduced frequency of sinus node. Both average heart rate and average minimum heart rate of the patients at night (74.0 +/- 5.6 BPM and 57.0 +/- 5.1 BPM, respectively) decreased significantly, as compared with controls (84.6 +/- 9.3 BPM and 66.3 +/- 8.0 BPM respectively) (p < 0.01). CVRR of the patients (0.120 +/- 0.019, respectively) increased significantly as compared with controls (0.090 +/- 0.010, respectively). These results indicate that an activated parasympathetic nervous system, that is, activation of the vagal nerve, might have induced the sinus bradycardia observed in the patients with EHEC infection.

A rapid method for starting a culture for the establishment of Epstein-Barr virus-transformed human lymphoblastoid cell lines.

We developed a simple and efficient procedure for the establishment of Epstein-Barr virus-transformed human lymphoblastoid cell lines. B-lymphocytes were obtained by centrifugation after hemolysis of red cells with a hemolysis buffer, instead of Ficoll-Parque gradient. We can start a primary culture within 15 min by using this method.

The interaction of interleukin 2 with its receptor in the generation of suppressor T cells in antigen-specific and antigen-nonspecific systems in vitro.

The role of interleukin-2 (IL-2) in the activation of suppressor T cells was investigated using the monoclonal antibody anti-Tac, which blocks the binding of IL-2 to the 55-kDa IL-2-binding peptide. The addition of anti-Tac during a preculture period inhibited the generation of Epstein-Barr virus (EBV)-induced suppressor T cells and of suppressor T cells induced in an antigen-specific system by a high antigen (sheep red blood cell) concentration. The cells activated by a short 2- or 7-day preculture period with EBV to become suppressor effectors were of the T8, Tac-positive phenotype. However, the T8-positive T cells obtained from peripheral blood mononuclear cells precultured with EBV for 14 days continued to manifest suppressor function, even though they were no longer Tac positive. In summary, our studies indicate that anti-Tac, by producing a functional blockade of human IL-2 receptors, inhibits the generation of suppressor T cells in antigen-specific as well as antigen-nonspecific systems and that cells that no longer express the Tac antigen may also function as suppressors.

Blockade of the interleukin-2 receptor by anti-Tac antibody inhibits the generation of antigen-nonspecific suppressor T cells in vitro.

The role of interleukin 2 (IL-2) in the activation of suppressor T cells was investigated by using the monoclonal antibody anti-Tac, which blocks the binding of IL-2 to the 55-kDa peptide of the high-affinity IL-2 receptor. Anti-Tac was added to an antigen-nonspecific suppressor system in which Con A-induced suppressor T cells were generated during a preculture period, and their effects on immunoglobulin production were assessed in second, indicator cultures containing pokeweed mitogen and peripheral blood mononuclear cells. Anti-Tac added during the preculture period inhibited Con A-induced suppressor T-cell generation. Cells activated by a short (2-day) preculture period to become effectors of suppression were primarily of the Tac-positive, T8 (CD8)-positive phenotype. Tac-positive, T8-negative T cells might also contribute to the suppressor activity. Our studies indicate that anti-Tac, by producing a functional blockade of human high-affinity IL-2 receptors, inhibits the generation of antigen-nonspecific suppressor T cells.