High Epstein-Barr virus serum load and elevated titers of anti-ZEBRA antibodies in patients with EBV-harboring tumor cells of Hodgkin's disease

Hodgkin's disease is commonly associated with EBV latent infection. The incidence of EBV reactivation (active infection or EBV infection with replicative cycle) was evaluated in a series of 30 patients with untreated Hodgkin's disease (except for one case with chronic lymphocytic leukemia) by quantitation of EBV DNA and titration of anti-ZEBRA antibodies in serum samples. DNA was detected in serum (>2.5 x 10(2) genomes/ml) in 15 of 30 patients and was more frequent in Hodgkin's disease with EBV-positive Reed-Sternberg cells (10/12) than in EBV-negative cases (5/18), (P< 0.01). Of interest was the demonstration that viremia correlated well with increased titers of anti-ZEBRA IgG and/or standard serological profiles of EBV reactivation (12/15), (P < 0.05). However the lack of EBV replicative cycle in Reed-Sternberg cells (negative for ZEBRA antigen and early antigen BHLF1) suggests that the viral replication occurs in a nonneoplastic cell compartment rather than in tumor cells. The measurement of EBV DNA loads and the titration of anti-ZEBRA antibodies shed new lights on the link between activation of EBV replication and Hodgkin's disease: these serological markers together with the determination of the EBV status of the tumor suggest that replication of the viral genome occurs with a decreased efficiency of the immune system, thus allowing progression of the tumor.     

Elevated antibody titers to Epstein-Barr virus and low natural killer cell activity in patients with Chediak-Higashi syndrome

Four Venezuelan patients with the autosomal recessive Chediak-Higashi syndrome (CHS) were studied. The results confirm the severe reduction in natural killer (NK) cell activity, as previously described and showed also a decline in the activity of cells involved in antibody-dependent cellular cytotoxicity (ADCC). No defect was found in the production of immunoglobulins and of specific antibodies to measles, varicella, herpes simplex, and cytomegalo viruses. Two of the patients had extremely high antibody titers to the Epstein-Barr virus (EBV) specific viral capsid antigen (VCA), to the restricted (R) component of the EBV-induced early antigen complex, and to the EBV-associated nuclear antigen (EBNA). These two patients had enlarged livers, spleens, and lymph nodes indicative of the lymphoproliferative phase. The other two patients were initially negative for all EBV-associated antibodies but seroconverted subsequently and, in the course of a year, also developed high antibody titers to VCA and R. In one of these patients the primary infection was accompanied by moderate signs of infectious mononucleosis (IM) followed after more than 6 months by persistent hepatosplenomegaly. The other patient also developed signs of a lymphoproliferative syndrome with hepatosplenomegaly and jaundice and died 8 months later. Such high anti-R titers are seen frequently in Burkitt's lymphoma, but rarely in other conditions. It is likely that the high antibody titers reflect an increased production of VCA and R due to defective NK and ADCC cell activities so that productively infected B lymphocytes are no longer eliminated before they have synthesized maximal amounts of antigens. The high anti-EBNA titers suggest normal T lymphocyte function. The possibility that the accelerated, lymphoma-like phase of the CHS involves EBV-transformed cells is discussed.     

Sinus histiocytosis with massive lymphadenopathy (Rosai-Dorfman disease) in a patient with elevated Epstein-Barr virus titers.

Sinus histiocytosis with massive lymphadenopathy (SHML) is a newly recognized disorder. The etiology of this disease is unknown. An exaggerated response to an offending agent such as the Epstein-Barr virus or Klebsiella bacteria has been postulated. Its course is usually benign. Cervical adenopathy is seen in 97% of the patients, while 30% of patients have nodal involvement in other sites, and 30% have extranodal involvement. There is a 7% mortality rate that occurs primarily in patients with immunologic defects. Corticosteroids ameliorate the constitutional symptoms, but cyclophosphamide appears to have the most beneficial effect. This article presents the case of a patient with SHML who demonstrated elevated Epstein-Barr virus titers.     

High incidence of Epstein-Barr virus genomes in Hodgkin's disease.

Tissue specimens from 198 cases of Hodgkin's disease and 151 non-Hodgkin lymphomas, as well as 34 nonmalignant lymph node biopsies were examined for the presence of Epstein-Barr virus (EBV) DNA by polymerase chain reaction. Epstein-Barr virus-specific DNA sequences were detected in DNA extracts from frozen and/or paraffin-embedded tissues of 58% of Hodgkin's disease cases. High and low grade non-Hodgkin lymphomas as well as chronic lymphocytic leukemia biopsies contained EBV DNA in 26%, 14%, and 7% of the cases, respectively. Ten percent of the control lymph nodes with normal histology were EBV positive. In Hodgkin's disease biopsies, subsequent in situ hybridization revealed an exclusive localization of the viral DNA in the tumor cells. These findings suggest an involvement of EBV in the pathogenesis of Hodgkin's disease in a substantial proportion of cases.     

Antinuclear antibodies and elevated anti-Epstein-Barr virus titers in cancer patients.

One and one-half percent of human sera from patients seen at a clinic for treatment of cancer contained antibodies to the nuclei of chick kidney cells by indirect immunofluorescence tests. In the group of sera containing antinuclear antibodies, the geometric mean titer to Epstein-Barr virus (EBV) capsid antigen was significantly elevated. Sera obtained from normal adults or from patients with similar histological types of tumors that possessed no antinuclear antibodies contained lower levels of anti-EBV antibodies. The elevated titers to EBV were correlated with the presence or absence of antinuclear antibodies and not with a particular type or site of neoplastic disease.     

Determination of immune status in patients with low antibody titers for rubella virus.

Three assays for detection of rubella antibodies, Rubella G (fluorescence immunoassay [FIA]), Rubacell (passive hemagglutination), and Rubaquick (passive hemagglutination with rotation), were compared with hemagglutination inhibition. A total of 100 serum specimens were selected, 68 of which had an FIA value of less than or equal to 25. On initial testing, among the four tests, there was agreement for 88 specimens for assignment of rubella immune status. On repeat testing, all the results agreed by the hemagglutination inhibition, passive hemagglutination, and passive hemagglutination rotation methods, and only one discrepant specimen remained by FIA.     

Danish children in Greenland have high Epstein-Barr virus titers.

Danish children brought to Greenland have somewhat higher mean titers of antibodies to Epstein-Barr virus than do Danish children in Denmark, although their titers are lower than those found in Eskimo children. This study presents the first evidence of increases in Epstein-Barr virus titers in migrant children.     

Low incidence and high titers of antibodies to hepatitis B virus X-protein in sera of Chinese patients with hepatocellular carcinoma

Sera of patients from China with hepatocellular carcinoma (HCC) were tested for the presence of HBc/HBe- and HBx antibodies by immunoblotting using recombinant MS2 or beta gal fusion proteins as substrate. Antibodies against HBx were detected in four out of 68 HBsAg positive and in one out of three HBsAg negative sera, antibodies against HBc/HBe in 52 and two serum samples, respectively. Competition experiments in which sera were preincubated with individual viral proteins synthesized in E. coli were carried out to demonstrate the specificity of signals obtained in immunoblot analyses. In the five anti-HBx positive sera, the antibody titer against X fusion protein was higher than against core fusion protein and in one of these sera anti-x activity could be demonstrated even at a serum dilution of 1:50,000. These data indicate that X antibodies occur rarely in Chinese patients and are not serodiagnostic for HCC. The high titer of X antibodies in some patients shows that the X protein can be highly immunogenic in vivo. Induction of antibody formation may be triggered by X protein expressed from integrated viral DNA.     

Antibodies to Epstein-Barr virus in patients with cryptococcosis.

Antibody levels to the Epstein-Barr virus, the etiological agent for heterophile-positive infectious mononucleosis, have been demonstrated in high titer in a number of lymphomas as well as infectious mononucleosis. Recent reports have suggested that the elevated antibody levels to Epstein-Barr virus may be the nonspecific result of disordered cell-mediated immunity. This study of patients with cryptococcosis was therefore undertaken to examine another disorder of known etiology associated with a defect in cell-mediated immunity. In this study we found that antibody levels in cryptococcosis patients, including a group specifically demonstrated to be anergic to a series of skin test antigens, were no different than those in matched normal controls. At the present time, therefore, it is unlikely that elevated antibody levels can be explained solely on the basis of depressed cellular immunity.     

Sinus histiocytosis with massive lymphadenopathy in a Japanese adult with unusually elevated EBV antibody titers

A 35-year-old Japanese male was presented with massive lymphadenopathy in bilateral preauricular and submandibular regions for five years' duration without any complaints despite of no specific treatments. Laboratory examinations revealed neurtrophilia, elevated BSR, hypergammaglobulinemia with elevated IgG and IgM, positive CRP and RA, low percentage of T-cells in peripheral blood, impaired PHA blast transformation, and elevated EBV titers against viral capsid, early and nuclear antigens. Biopsy specimens demonstrated massive histiocytosis with hemophagocytosis in the sinuses and predominant mature plasma cells in the medulla, which were fairly well consistent with "sinus histiocytosis with massive lymphadenopathy" (ROSAI and DORFMAN). Ultrastructural study revealed histiocytes exhibiting avid phagocytosis of blood cells, epithelioid histiocytes with poor phagocytic activity, foamy storage histiocytes loaded with a large number of osmiophilic lipid granules and giant cells of various types. Pathogenesis of this apparently benign disease entity was briefly discussed, and its refractoriness against any specific therapy was emphasized.     

High titers of cytopathic virus in plasma of patients with symptomatic primary HIV-1 infection

BACKGROUND. Primary infection with the human immunodeficiency virus (HIV-1) frequently causes an acute, self-limited viral syndrome. To examine the relations among viral replication, the immune response of the host, and clinical illness during this initial phase of infection, we undertook a quantitative, molecular, and biologic analysis of infectious HIV-1 in the blood and plasma of three patients with symptomatic primary infection and of a sexual partner of one of them. METHODS. During an eight-week period of primary infection, HIV-1 was cultured frequently in dilutions of plasma and peripheral-blood mononuclear cells (PBMC), and levels of HIV-1 antigen and antibody were determined sequentially by enzyme-linked immunosorbent assay and immunoblotting. Replication-competent HIV-1 proviruses were cloned and characterized biologically. RESULTS. Six to 15 days after the onset of symptoms, high titers of infectious HIV-1 (from 10 to 10(3) tissue-culture-infective doses per milliliter of plasma) and viral p24 antigen were detected in the plasma of all three patients. These titers fell precipitously by day 27, and the decline coincided with an increase in the levels of antiviral antibodies and the resolution of symptoms. Sequential isolates of virus from plasma and PBMC obtained throughout the period of primary infection, as well as virus derived from two molecular proviral clones, were highly cytopathic for normal-donor PBMC and immortalized T cells, despite the marked reduction in the titers of virus in plasma. CONCLUSIONS. Primary, symptomatic HIV-1 infection is associated with high titers of cytopathic, replication-competent viral strains, and during such infection potential infectivity is enhanced. Effective control of HIV-1 replication during primary infection implies the activation of clinically important mechanisms of immune defense that merit further examination in relation to the development of antiviral therapy and vaccines.     

High titers of anti-epstein-barr virus DNA polymerase are found in patients with severe fatiguing illness

Forty-one patients with chronic fatigue syndrome (CFS), 76 healthy controls matched with the patient group for age range, sex, race, and socioeconomic class, and 22 symptomatic patients with seasonal affective disorder (SAD) had serum sampled for antibodies against 2 Epstein-Barr virus (EBV) replicating enzymes. Abnormal liters of antibodies were found twice as often in CFS patients as controls (34.1% vs. 17.1%), with SAD patients having an intermediate frequency (27.3%). Stratifying for disease severity sharpened the differences considerably, with the sicker CFS and SAD patients having 52% and 50% abnormal tests, respectively; more mildly afflicted CFS and SAD patients had a frequency of abnormal tests in the normal range. Antibodies to EBV DNA polymerase (DNAP) were the more sensitive of the two tests in that they were positive in all cases but one. These findings suggest that antibodies against EBV DNAP may be a useful marker in delineating a subset of patients with severe fatiguing illness for appropriate treatment trials and for monitoring their outcomes. © 1994 Wiiey-Liss, Inc.     

Enhancement of Epstein-Barr virus antibody production in systemic lupus erythematosus patients.

Two groups of 22 patients suffering from either systemic lupus erythematosus (SLE) or infectious mononucleosis (IM) were checked for Epstein-Barr virus capsid antigen antibody (EB-VCA) production. The average significant antibody levels as well as the frequency of their occurrence were clearly higher in SLE than in IM patients.     

A screening test for influenza immunity: preimmunization antibody titers to influenza virus antigens in atopic patients

Antibody titers to three common strains of influenza virus, including the Swine Flu strain, were determined in 152 patients in an allergy and pediatric allergy practice prior to planned immunization with influenza vaccine. Because allergic patients are considered to be more prone to hypersensitivity reactions than individuals who are non-allergic, it was deemed prudent to screen for patients who might have pre-existing protective levels of antibody to the viruses. A considerable number of the 152 patients were found to have titers of 1:20 or 1:40 to one or more of the three influenza strains, including the Swine Flu strain. The largest percentage of positive antibody levels was found to be to the A-Victoria strain which has recently been most prevalent in the northeastern United States. The lowest numbers of patients with protective antibody levels were for the Swine Flu strain. Nevertheless, the highest titers occurred with patient specimens tested against the Swine Flu strain. Approximately one-quarter of the patients showed significant levels of antibody (1:40 or more) to one or more of the virus antigens and thus were not vaccinated, avoiding the possibility of untoward areactions which habe been observed in some individuals who have been given the vaccine.     

High incidence of peripheral blood plasmacytosis in patients with dengue virus infection

Little is known about polyclonal peripheral blood plasmacytosis in dengue virus (DENV)-infected patients. We initiated this prospective observational study to quantify and describe the kinetics and phenotype of peripheral blood plasma cells (PCs) in these patients. Morphological examination and flow cytometric (FC) analysis for the characterization and immunophenotyping of lymphocyte subsets and PCs were performed in 35 and 31 patients suspected of DENV infection, respectively. Our results show that blood plasmacytosis is a very common haematological finding. Depending on the days of illness at presentation, blood plasmacytosis was observed in 64% to 73% of patients. Blood plasmacytosis was most pronounced before 7 days of illness and declined rapidly thereafter, to completely disappear after 14 days of illness. Blood plasmacytosis was higher in secondary DENV infection. The majority of CD138⁺ PCs (89%) had a shared immunophenotype (CD45⁺/CD19^-/CD56^-) and in all cases the PCs were polyclonal. Blood plasmacytosis, characterized by a transient presence of polyclonal PCs in the circulation, is a common event in DENV infection.     

Detection of Epstein-Barr virus specific IgE antibody

To confirm the existence and investigate the biological significance of IgE virus-specific antibodies, we studied Epstein-Barr virus (EBV)-specific IgE antibody by enzyme-linked immunosorbent assay with an anti human IgE monoclonal antibody. We detected EBV-specific IgE antibody in sera not only from patients with the EBV associated diseases of infectious mononucleosis and nasopharyngeal carcinoma, but also from patients with bronchial asthma, collagen disease and healthy volunteers. However, there was no significant difference in the titers of IgE antibody specific for EBV among these groups. No significant relationship between the titers of EBV-specific IgG and IgE antibody, or between the titers of EBV-specific IgE and the total IgE levels in the sera was observed.     

Herpes simplex virus, cytomegalovirus and Epstein-Barr virus antibody titres in sera from schizophrenic patients

Serum antibody titres to herpes-simplex (HSV-1, 2), cytomegalovirus (CMV), and Epstein-Barr virus capsid antigen (EBV-VCA) were determined in 38 unrelated chronic schizophrenic patients, 11 nuclear families with at least 2 schizophrenic members, and 2 control groups. The distributions of antibody titres to herpes simplex and cytomegalovirus were similar among all groups. Patients had higher anti-EBV-VCA titres than non-hospitalized controls; however, hospital staff members in contact with the patients also had significantly higher antibody titres to EBV-VCA. Antibodies to EBV early antigen (EBV-EA) were also determined for 27 unrelated patients and 24 mental hospital employees. The schizophrenic patients had significantly higher antibody titres to EBV-EA than the hospital worker control group. These data do not support the hypothesis that herpes viruses are associated with the aetiology of schizophrenia. Although elevated anti-EBV early antigen titres may suggest persistent active EBV infection, it is unlikely to be related to the aetiology of the disorder, since discordance for EBV seropositivity was present among sibling pairs concordant for schizophrenia.     

Response to valganciclovir in chronic fatigue syndrome patients with human herpesvirus 6 and Epstein–Barr virus IgG antibody titers

Valganciclovir has been reported to improve physical and cognitive symptoms in patients with chronic fatigue syndrome (CFS) with elevated human herpesvirus 6 (HHV‐6) and Epstein–Barr virus (EBV) IgG antibody titers. This study investigated whether antibody titers against HHV‐6 and EBV were associated with clinical response to valganciclovir in a subset of CFS patients. An uncontrolled, unblinded retrospective chart review was performed on 61 CFS patients treated with 900 mg valganciclovir daily (55 of whom took an induction dose of 1,800 mg daily for the first 3 weeks). Antibody titers were considered high if HHV‐6 IgG ≥1:320, EBV viral capsid antigen (VCA) IgG ≥1:640, and EBV early antigen (EA) IgG ≥1:160. Patients self‐rated physical and cognitive functioning as a percentage of their functioning prior to illness. Patients were categorized as responders if they experienced at least 30% improvement in physical and/or cognitive functioning. Thirty‐two patients (52%) were categorized as responders. Among these, 19 patients (59%) responded physically and 26 patients (81%) responded cognitively. Baseline antibody titers showed no significant association with response. After treatment, the average change in physical and cognitive functioning levels for all patients was +19% and +23%, respectively (P < 0.0001). Longer treatment was associated with improved response (P = 0.0002). No significant difference was found between responders and non‐responders among other variables analyzed. Valganciclovir treatment, independent of the baseline antibody titers, was associated with self‐rated improvement in physical and cognitive functioning for CFS patients who had positive HHV‐6 and/or EBV serologies. Longer valganciclovir treatment correlated with an improved response. J. Med. Virol. 84:1967–1974, 2012. © 2012 Wiley Periodicals, Inc.