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PubMed2005

Multiple Epstein-Barr virus strains in patients with infectious mononucleosis: comparison of ex vivo samples with in vitro isolates by use of heteroduplex tracking assays.

Recent work using a heteroduplex tracking assay (HTA) to identify resident viral sequences has suggested that patients with infectious mononucleosis (IM) who are undergoing primary Epstein-Barr virus (EBV) infection frequently harbor different EBV strains. Here, we examine samples from patients with IM by use of a new Epstein-Barr nuclear antigen 2 HTA alongside the established latent membrane protein 1 HTA. Coresident allelic sequences were detected in ex vivo blood and throat wash samples from 13 of 14 patients with IM; most patients carried 2 or more type 1 strains, 1 patient carried 2 type 2 strains, and 1 patient carried both virus types. In contrast, coresident strains were detected in only 2 of 14 patients by in vitro B cell transformation, despite screening >20 isolates/patient. We infer that coacquisition of multiple strains is common in patients with IM, although only 1 strain tends to be rescued in vitro; whether nonrescued strains are present in low abundance or are transformation defective remains to be determined.

Blood↗
PubMed1981

Infectious mononucleosis: sequential immunologic, cytochemical, and cytokinetic studies on single lymphoid cells in peripheral blood.

Sequential immunologic, cytochemical, and cytokinetic studies were done on single lymphoid cells in the peripheral blood of 6 children with infectious mononucleosis (IM) and 1 child with toxoplasmosis 1-2 wk after onset of symptomatology. The absolute number of AET-SRBC rosetting (E+) cells was increased in all patients; the absolute number of cells with surface IgM (slgM+) was increased in 3 of 7 patients. On the average, 46% of all lymphoid cells were E+la+slgM-, 35% E+la-slgM-, and 5% E-la+slgM+. Of the E+la+slgM- cells, only 7% were shown to have a dot-like esterase reaction pattern. Of the E+la-slgM-cells, 17% were esterase positive. The mean labeling index of the former was 18.2%, the mean labeling index of the latter 4.4%. No proliferating slgM+ cells were observed in the 6 children with IM. The esterase reactions and the labeling indices of suppressor-cytotoxic and inducer-helper T cells were similar to those of E+la+slgM- and E+la-slgM- cells, respectively. These results suggest that the vast majority of the atypical cells in IM corresponds to activated suppressor cells, while a minority appears to be proliferating helper cells.

Child↗
PubMed1998

A boy with fatal infectious mononucleosis suspected as the first Japanese case of X-linked lymphoproliferative syndrome.

We report a case of a 10-month-old boy who died of severe hepatic failure after a prolonged course of infectious mononucleosis. He also presented interstitial pneumonitis, meningoencephalitis and aplastic anaemia. Epstein-Barr virus (EBV)-specific cytotoxic T lymphocyte (CTL) activity had not been detected in his peripheral blood during the course of the illness. Studies of his mother revealed a severe reactivation pattern of anti-EBV antibodies and decreased EBV-specific CTL activity. An X-linked familial susceptibility to EBV infection such as X-linked lymphoproliferative syndrome (XLP) might be associated with his fatal EBV infection.

Cytotoxicity, Immunologic↗
PubMed1989

Identification of Epstein-Barr virus-infected cells in tonsils of acute infectious mononucleosis by in situ hybridization.

In situ hybridization with 35S-labeled Epstein-Barr virus (EBV) probes was applied to paraffin sections of tonsils from seven patients with clinical, serologic, and morphologic evidence of acute infectious mononucleosis. EBV genomes were demonstrated in activated lymphoid B blasts in the interfollicular and perifollicular zones in all these cases. However, in no case could EBV be identified in epithelial cells. These results are at variance with the current concept which attributes a central role to the tonsillar epithelium in primary EBV infection.

Acute Disease↗
PubMed1992

[A new modification of the horse erythrocyte agglutination test for the detection of heterophilic antibodies in infectious mononucleosis].

The new modification (HEA test) is based on the presence of a singularly distinct agglutination of native horse erythrocytes in a serum dilution 1:10 and more from cases with Epstein-Barr virus infectious mononucleosis. In 1665 sera the HEA test was equally often positive as the OCH test (in 5.2%) whereas the IM test with formal-treated horse erythrocytes was positive only in 3.7%. The sensitivity of the IM test in comparison with the OCH test was 61%, the specificity 99.5%. The sensitivity of the HEA test in comparison with the OCH test was 76%, the specificity 98.7%. The specificity of the HEA test in a group of 52 sera without IgG antibodies to EB-viral capsid antigen (EB VCA) was 94%, in another group of 130 sera from hepatitis cases it was 98%. In 103 pairs of sera from infants aged 1-5 years the positivity of the OCH test, HEA test, IgM antibodies to EB VCA and the four-fold rise in the titre of EB-viral IgG were, 20%, 16%, 23% and 24% respectively. Analogous values in 159 pairs of sera from older subjects were 42%, 40%, 38% and 29%. The HEA test is recommended as useful adjunct to the OCH test for the detection of heterophile antibodies even in young children.

Agglutination Tests↗
PubMed1990

Immunovirological studies of fatal infectious mononucleosis in a patient with X-linked lymphoproliferative syndrome treated with intravenous immunoglobulin and interferon-alpha.

We have studied a 19-year-old male with X-linked lymphoproliferative syndrome (XLP) and infectious mononucleosis (IM) who was treated with high-dose immunoglobulin (500 mg/kg/day) and recombinant interferon (IFN)-alpha (2 x 10(6) IU/m2/day). Fulminant hepatitis was delayed; however, virus-associated hemophagocytic syndrome, cholestatic jaundice, and renal failure occurred terminally. Initially, nonspecific natural killer (NK) cell activity against K562 cells was normal but it gradually decreased. Although reactive T cells were markedly increased in his blood during the acute phase, spontaneous EBV-positive cell lines were easily established. Additionally, his mononuclear cells produced IFN-gamma but not IFN-alpha prior to treatment. Based on results of in vitro studies, we conclude that both IFN-alpha and IFN-gamma production are likely necessary for inhibiting EBV immortalization in vitro. Both IFN-alpha and -gamma were produced in cultures of B95-8 EBV-infected mononuclear cells from EBV-seropositive healthy individuals. These results suggest that defective EBV-specific cytotoxic T cell activity accompanied with defective or discordant IFN-alpha and -gamma production permitted the development of fatal IM in this patient. Combined treatment with immunoglobulin and IFN-alpha appeared to be partially effective during the early stage of this disease.

Adult↗
PubMed1982

Evolution of infectious mononucleosis into Epstein-Barr virus carrying monoclonal malignant lymphoma.

A monoclonal malignant lymphoma was diagnosed in a 30-month-old Japanese boy, who, since the age of 12 months, had had chronic, recurrent infectious mononucleosis manifested by repeated episodes of severe cough and high fever accompanied by marked lymphadenopathy and hepatosplenomegaly and high serum Epstein-Barr virus (EBV) antibody titres. The diagnosis of Burkitt's lymphoma was made when a cervical lymph-node biopsy specimen revealed massive proliferation of immature B-cells with starry-sky histiocytes. These lymph-node cells were characterised by a translocation between chromosomes 10 and 17. There were approximately 9 EBV genome-equivalents per cell. Most cells were positive for nuclear antigen (EBNA); early antigen (EA) and viral capsid antigen (VCA) were also detected. The presence of EBV was supported by finding that the cell-free lymph-node extract transformed cord-blood lymphocytes into EBNA-positive blast cells. The proportion of EA-positive and VCA-positive cells increased rapidly in culture for 24 h, then the positive cells degenerated rapidly and completely. The cells also contained numerous herpes-type virus particles. The child improved considerably with cytostatic treatment and has been in remission for 2 years.

Antibodies, Viral↗
PubMed1980

Studies of lymphocytotoxins in infectious mononucleosis and systemic lupus erythematosus: evidence for immune complex-mediated cytotoxicity.

The hypothesis that serum lymphocytotoxins are antigen-antibody complexes was examined. High molecular weight fractions from the sera of eighteen patients with infectious mononucleosis (IM), thirteen patients with systemic lupus erythematosus (SLE) and six healthy controls, were prepared by precipitation with polyethylene glycol 6000 (PEG). The lymphocytotoxic activity (LCA) of these PEG precipitates was significantly greater (P less than 0.01) than that of the corresponding sera and a significant correlation (r = 0.66, P less than 0.01) was observed between the LCA of sera and the PEG precipitates. In contrast to the concentration of LCA in the PEG precipitates, the heterophil antibody titres of the precipitates from IM sera were significantly less (P less than 0 05) than serum titres. Antisera raised against PEG precipitates from sera from nine patients with IM contained significant LCA. The nature of this LCA differed from that of the LCA in the original sera in temperature dependence and the molecular size. Antigen-antibody complexes in seven sera (four IM, three SLE) were dissociated at low pH (3.0) and fractionated by gel filtration at pH 3.9. The LCA of these fractions was compared with the LCA of equivalent fractions obtained by gel filtration at pH 7.2. The heterophil antibody present in sera from patients with IM and the cytotoxicity of anti-lymphocyte globulin (ALG) were used as 'antibody controls'. In this way it was shown that the LCA in patient sera, but not heterophil antibody or ALG cytotoxicity was significantly reduced (P less than 0.001) by low pH gel filtration.

Antigen-Antibody Complex↗
PubMed2005

Long-term shedding of infectious epstein-barr virus after infectious mononucleosis.

Epstein-Barr virus (EBV) DNA loads in peripheral blood mononuclear cells (PBMCs), plasma, and saliva, as well as infectivity of the virus in saliva, were evaluated in 20 patients for 6 months after the onset of infectious mononucleosis (IM). All patients displayed sustained high EBV DNA loads in the saliva, associated with a persistent infectivity of saliva at day 180. EBV DNA load in PBMCs decreased significantly from day 0 to day 180 (in spite of a viral rebound between day 30 and day 90 in 90% of the patients), and EBV DNA rapidly disappeared from plasma. These data show that patients with IM remain highly infectious during convalescence.

Adolescent↗
PubMed1990

[Heterophile infectious mononucleosis antigen used in the latex diagnostic test. I. A method of antigen isolation and its serologic activity].

The aim of this study was to elaborate a method of heterophile mononucleosis antigen preparation useful for latex coating. This antigen was isolated from bovine red blood cells stroma by the technique of Schwarzweiss and Tomcsik with author's own modification, in which introductory extraction of erythrocytes stroma ++ was performed by means of trichloracetic acid, aqueous extraction and elution of active substance with 80% ethanol. Besides of heterophile antigen preparation obtained by the method of Schwerzweiss and Tomcsik (preparation S-T) two serologically++ active preparations were obtained (fraction I and IV), which ability to inhibit PBD agglutinating reaction and bovine red blood cells haemolysis was 16 and 8 times lower, respectively, than S-T preparation. The preparation of heterophile mononucleosis antigen obtained differed in latex coating efficacy. In order to prepare latex reagent MZ-I (from fraction I) a solution of preparation of 125 micrograms/ml concentration was used, for MZ-II (from fraction IV)--50 micrograms and for MZ-III (from preparation S-T)--15 micrograms/ml. The reagent MZ-I showed, the highest activity in agglutinating test with human serum containing heterophile mononucleosis antibodies while two others reacted with 2-4 times lover serum dilutions. Similar differentiated reactivity with these reagents was found in latex test with 15 sera from patients suspected of having infectious mononucleosis.

Animals↗
PubMed2003

Epstein-Barr virus-associated infectious mononucleosis in Chinese children.

BACKGROUND: According to seroprevalence studies the majority of children in Hong Kong are infected by Epstein-Barr virus (EBV) before 10 years of age, but the characteristics of EBV-associated infectious mononucleosis (IM) in Chinese children are largely unreported. This study aims at defining the clinical presentation and complications of Chinese childhood IM in relation to age of the children. METHODS: A retrospective study was performed on 77 consecutive Chinese childhood IM patients who fulfilled the serologic criteria for the diagnosis of primary EBV infection (viral capsid antigen IgM+ viral capsid antigen IgG+ Epstein-Barr nuclear antigen-). The clinical, hematologic and biochemical findings were evaluated among four age groups of <2 years, 2 to 4 years, 5 to 9 years and 10 to 15 years. RESULTS AND CONCLUSIONS: EBV-associated IM occurred at all age groups with a peak incidence at 2 to 4 years, corresponding to the rapid rise in the seroprevalence of EBV in early childhood in the Hong Kong Chinese. The majority of children presented with fever, tonsillopharyngitis, lymphadenopathy and hepatosplenomegaly, similar to the adult IM patients, and recovered without major complications. Marked lymphocytosis with the presence of atypical lymphocytes was a consistent hematologic finding in all age groups. The occurrence of hepatitis showed a clear association with advancing age (P = 0.003). The age-related increase in IM-associated hepatitis may reflect difference in the host immune response against EBV between the infants and older children.

Adolescent↗
PubMed1980

Sudden death due to spontaneous rupture of the spleen from infectious mononucleosis.

An 18-year-old white male showing no external signs of trauma was discovered dead in his bathroom. A ten-day history of illness resulting in symptomatic treatment for viremia one day before death was elicited. Autopsy revealed splenic rupture, massive peritoneal hemorrhage, and atypical lymphocytes. Laboratory tests on postmortem samples confirmed the diagnosis of infectious mononucleosis.

Adolescent↗
PubMed1982

Detection of infectious mononucleosis heterophil antibody by a rapid, standardized enzyme-linked immunosorbent assay procedure.

A rapid, specific, sensitive, standardized, a reproducible enzyme-linked immunosorbent assay (ELISA) procedure has been developed for detecting the heterophil antibody associated with infectious mononucleosis (IM). The IM heterophil antibody used for the solid phase was purified from bovine erythrocyte stroma. The test uses heavy-chain-specific anti-immunoglobulin M (IgM) labeled with alkaline phosphatase and three 10-min incubations. The quantitative results correlated well with horse erythrocyte agglutination titers. Absorption tests confirmed the specificity of the ELISA reactions for IM heterophil antibodies. Neither very high levels of IgM in myeloma sera nor high levels of rheumatoid factor caused false-positive reactions. A number of probable IM cases were encountered which positive by ELISA but negative by the horse erythrocyte slide agglutination test. Absorption studies indicated that these were true-positives for the IM heterophil antibody. The IM heterophil antibodies were confirmed to be predominantly of the IgM class, but moderate proportions of the IgM class were sometimes encountered.

Antibodies, Viral↗
PubMed1991

Expression of CD45R0 (UCHL1) by CD4+ and CD8+ T cells as a sign of in vivo activation in infectious mononucleosis.

CD45R0 (UCHL1), a member of leucocyte common antigen family, is expressed largely on previously activated or memory T cells. We examined CD45R0 expression of T cell subpopulations in patients with Epstein-Barr virus (EBV) induced infectious mononucleosis (IMN) as a sign of in vivo activation. Consistent with the notion that activated CD8+ T cells expand in acute IMN; the majority of CD8+ T cells in patients with acute IMN expressed CD45R0 to the similar extent to HLA-DR expression. Most CD4+ T cells in these patients also demonstrated marked expression of CD45R0 as well as HLA-DR antigens, compared with age-matched controls. Expression of CD45R0 by CD4+ T cells in patients with acute IMN was more notable than their HLA-DR expression. While predominant CD8+ T cells resulted in decreased percentages of CD4+ T cells, CD4+ T cells expressing CD45R0 were shown to be significantly elevated in absolute number. The results suggest that both CD4+ and CD8+ T cells may be activated by stimulation with EBV infection. The appearance of two T cell subpopulations expressing CD45R0 in acute IMN implies their immunoregulatory roles in the control of EBV-infected cells.

Antigens, CD↗
PubMed1996

Direct detection of Epstein-Barr viral antigen in nasopharyngeal swabs from patients with infectious mononucleosis.

OBJECTIVE: To determine whether direct immunologic detection of Epstein-Barr virus (EBV) early antigen diffuse component (EA-D) from nasopharyngeal swabs is a feasible approach to the development of a rapid diagnostic test for infectious mononucleosis (IM). METHODS: Nasopharyngeal swabs from 20 patients presenting with a presumptive diagnosis of IM (having the classic triad of symptoms-acute pharyngitis, fever, and lymphadenopathy) and 5 controls were assayed for EA-D. EBV serologic testing and a heterophil antibody titer (HAT) test also were performed. EA-D was assayed by polyacrylamide gel electrophoresis and subsequent transfer to a nylon membrane, followed by immunoblotting with a monoclonal antibody. RESULTS: EA-D was detected in 17 of 20 patients (85%) with presumptive diagnoses of IM and in 1 of 5 normal subjects and was highly significant in predicting IM (p < 0.01). There was no significant difference in numbers of positive and negative results using either EA-D assay or HAT test in patients with IM (p < 0.35). Pharyngeal exudate in the 17 pharyngitis patients with this variable documented was significantly correlated with positive EA-D (p < 0.01), but not with the HAT test (p < 1.00). CONCLUSIONS: Immunologic detection of EBV-derived antigens from nasopharyngeal swabs is a potential early diagnostic tool for clinically suspected IM. Sensitivity and specificity in pediatric and adult populations, patients with other viral etiologies, and patients with streptoccocal pharyngitis should be determined in subsequent investigations.

Adolescent↗
PubMed2004

T cell reactivity during infectious mononucleosis and persistent gammaherpesvirus infection in mice.

Intranasal infection of mice with murine gammaherpesvirus 68 causes a dramatic increase in numbers of activated CD8(+) T cells in the blood, analogous in many respects to EBV-induced infectious mononucleosis in humans. In the mouse model, this lymphocytosis has two distinct components: an early, conventional virus-specific CD8(+) T cell response, and a later response characterized by a dramatic increase among CD8(+) T cells that bear Vbeta4(+) TCRs. We previously demonstrated that Vbeta4(+)CD8(+) T cells recognize an uncharacterized ligand expressed on latently infected B cells in an MHC-independent manner. The frequency of Vbeta4(+)CD8(+) T cells increases dramatically following the peak of viral latency in the spleen. In the current studies, we show that elevated Vbeta4(+)CD8(+) T cell levels are sustained long-term in persistently infected mice, apparently a consequence of continued ligand expression. In addition, we show that Vbeta4(+)CD8(+) T cells can acquire effector functions, including cytotoxicity and the capacity to secrete IFN-gamma, although they have an atypical activation profile compared with well-characterized CD8(+) T cells specific for conventional viral epitopes. The characteristics of Vbeta4(+)CD8(+) T cells (potential effector function, stimulation by latently infected B cells, and kinetics of expansion) suggested that this dominant T cell response plays a key role in the immune control of latent virus. However, Ab depletion and adoptive transfer studies show that Vbeta4(+)CD8(+) T cells are not essential for this function. This murine model of infection may provide insight into the role of unusual populations of activated T cells associated with persistent viral infections.

Animals↗