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A prospective follow-up of Epstein-Barr virus LMP1 genotypes in saliva and blood during infectious mononucleosis.

To monitor multiple Epstein-Barr virus (EBV) infections during the early and convalescent stages of infectious mononucleosis (IM), a cloning and sequencing study of the LMP1 gene was conducted in saliva and peripheral blood mononuclear cells (PBMCs) from 23 patients with IM at day 0 (D0) and day 180 (D180) after the onset of the disease. Multiple EBV strains were detected in 9 (39%) of the patients during follow-up, with 7 of 9 cases detected as early as D0. Six of the nine patients harbored the same dominant strain in saliva and PBMCs during follow-up, with a trend toward a restriction of the number of EBV strains in saliva but not in PBMCs at D180. Furthermore, transmission of a minor strain was observed between partners in a heterosexual couple. There was no correlation between multiple infections and EBV DNA load in either compartment.

Cloning, Molecular↗

Infectious mononucleosis: increase of high-affinity E-receptor-bearing lymphocytes.

The increase of T lymphocytes in infectious mononucleosis (IM) is shown to be due to an increase of 'active' (early erythrocyte rosette-forming) T cells: 67.2 +/- 18.3% of IM lymphocytes were 'active' versus 34.7 +/- 14.9% of control lymphocytes. IM was also associated with an increase of lymphocytes demonstrating E receptors at 20 degrees C (E20-R). This phenomenon was not related to the heterophile antibody titre and could not be demonstrated with ox or rabbit erythrocytes. Patients with other lymphoproliferative diseases did not show an increase of E20-R-positive cells or of 'active' erythrocyte rosette-forming cells (E-RFC). Re-rosetting experiments indicated that E20-R and 'active' E-RFC belong to overlapping populations of cells. A double-gradient separation technique was shown to be superior to the conventional Ficoll-Isopaque method for the demonstration of 'active' and total T cells. A high proportion of T lymphocytes were also recovered in the granulocyte layer by this technique, indicating a selective loss of these cells by the conventional technique.

Cell Separation↗

Studies of lymphocytotoxins in infectious mononucleosis: reduced lymphocyte killing in the acute phase.

Fifty sera from twenty-five patients with uncomplicated infectious mononucleosis (IM) were tested for lymphocytotoxic activity (LCA) against pools of lymphocytes collected from (a) normal donors (b) ten patients during the acute phase of IM and (c) four of these patients during the convalescent phase. The LCA of twenty-four sera from patients with systemic lupus erythematosus (SLE) was also tested against the same panels of cells. The percentage of acute phase lymphocytes killed by sera from patients with IM or SLE was significantly less (P less than 0.001 and P less than 0.05 respectively) than the percentage of normal or convalescent cells killed by the same sera. This reduction could not be explained by lymphocyte donor variation as the same donors were used in the acute and convalescent phases of IM. It is suggested that changes in the surface characteristics of acute IM lymphocytes cause a temporary failure of interaction with circulating lymphocytotoxins. Such a phenomenon could have important influences on the self-limiting nature of the disease.

Acute Disease↗

Lymph node lesion in infectious mononucleosis showing geographic necrosis containing cytologically atypically B-cells. A case report.

Lymph node lesions in infectious mononucleosis (IM) show a marked histological diversity and may occasionally be confused with malignant lymphoma. We report on a rare case of IM showing geographic lymph node necrosis as well as angiocentric lymphoproliferative lesions, and containing numerous centroblasts, immunoblasts and Reed-Sternberg (RS)-like cells. The patient was a 40-year-old Japanese man with signs and symptoms of classical IM. This was later confirmed serologically, but the necrotic area comprised 50% of a cervical lymph node. The large lymphoid cells, including RS-like cells, were CD3-, CD5-, CD15-, CD20+, CD30+, CD45RO-, CD79a+, LMPI+, and EBNA2+. In situ hybridization study also disclosed that these cells were associated with Epstein-Barr virus (EBV). The patient was disease free during a follow-up of 15 years. Although the classical IM syndrome rarely shows a close resemblance to lymphomatoid granulomatosis of the lymph node or to EBV+ B- cell lymphoproliferative disorders associated with an immunodeficient state on histology, it is important for pathologists to be aware of this type of lesion in diagnostic practice.

Antigens, CD↗

Hepatosplenomegaly in infectious mononucleosis, assessed by ultrasonic scanning.

The present study aims at an assessment of hepato-splenomegaly in infectious mononucleosis (IM). In 29 patients admitted to the ENT department with IM, based on the typical clinical and laboratory findings, including a positive mononucleosis test in most cases, the size of the liver and spleen was estimated by ultrasonic scanning on days 1, 3, 5, 10, 20, 30, 90 and 120 after admission. A control group of eight patients with peritonsillar abscess was included for comparison. The results showed that all patients had an enlarged spleen (mean enlargement: 50-60 per cent) but only a few were palpable. Half of the patients had enlargement of the liver (5-20 per cent), which was palpable in only 8 per cent. There was no correlation between the size of the spleen and that of the liver, not between the changes in the size of these organs. There was no enlargement of the liver or spleen in the control group. No correlation was found between the size, or changes in the size, of the organs and blood values such as lactatdehydrogenase and aspartatamino-transferase. There is, however, a striking parallelism between the curves for these parameters, which might indicate that the organs as well as the blood tests return to normal within 28 days. If this holds true, our warning to abstain from physical exercise and alcoholic intake may be limited to a period of about 1 month.

Adolescent↗

Cellular immunity in infectious mononucleosis. II. Specific reactivity to Epstein-Barr Virus antigens and correlation with clinical and hematologic parameters.

Infectious mononucleosis (IM) patients, Epstein-Barr virus (EBV)-seropositive and seronegative healthy donors, and patients with other viral infections were tested for lymphocyte blastogenesis (LB) with phytohemagglutinin and six EBV (virus concentrate, culture supernatant, and soluble [S] antigen) or control antigens. Fluorescent antibodies to EBV viral capsid antigen of IgG, IgM, IgA specificities, to nuclear antigen (EBNA), and heterophile antibodies were also assayed. These were correlated with clinical parameters (fever, pharyngitis, adenopathy, hepatitis, splenomegaly, atypical lymphocytes, and total mononuclear cell counts). EBV viral and S antigen-induced LB was significantly greater in seropositive donors. IM patients had antigenspecific LB below that of seropositive donors initially and low responses for the acute phase of illness when clinical symptoms were present and antibody titers were maximal. Specific LB rose to a peak at 3.5 to 9 weeks when the patients had recovered, most laboratory findings had returned to normal, and antibodies had declined. At peak, specific LB in IM patients exceeded that of seropositive donors, but later declined. These results demonstrate specific cell-mediated immunity (CMI) to EBV, and indicate that this develops slowly in IM and contrasts with the evolution of the clinical events and humoral immunity. This correlation supports the hypothesis that CMI is the mechanism of terminating lymphoproliferation in IM.

Antibodies, Viral↗

T cell cytokine profile during primary Epstein-Barr virus infection (infectious mononucleosis).

Cytokine profiles of CD4+ and CD8+ T-cell subsets were evaluated in 8 patients with infectious mononucleosis (IM). Intracellular detection of cytokines using flow cytometry revealed an expansion of IFN-gamma-expressing CD4+ T cells, and particularly CD8+ T cells, while IL-2 expressing cells were less frequently encountered when compared to healthy controls. Single TNF-alpha-expressing CD4+ and CD8+ T cells were likewise reduced and shifted towards IFN-gamma/TNF-alpha co-production. The predominant pro-inflammatory type 1-biased immune response during IM was emphasized by low frequencies of IL-10 expression in both T cell subsets, although some patients displayed elevated serum levels. Six months later, a decreased, but still elevated IFN-gamma expression within the CD8+ T cell subset, and an increased percentage of IL-2-expressing CD4+ and CD8+ T cells, reaching values shown for controls, were noted. Type 2-associated cytokines such as IL-4 and IL-13, as well as IL-6 and TNF-alpha were not significantly different when compared to controls at study entry and at follow-up. The striking expansion of IFN-gamma-producing CD8+ T cells with rather low expression of IL-10, appears to be a key factor for clinically overt disease, but is nevertheless compatible with successful control of the viral infection.

CD4-CD8 Ratio↗

[The inhibition pathway of the EBV-immortalized cells in children with infectious mononucleosis].

OBJECTIVE: To explore the inhibition pathway of the EBV-immortalized cells (CD23(+)) in children with infectious mononucleosis (IM) caused by Epstein-Barr virus. METHODS: The expressions of CD23, CD19, CD95, Bcl-2 and the co-expressions of CD23CD95, CD19CD23 on peripheral blood mononuclear cell (PBMC) were analyzed by flow cytometry (FCM) during acute phase, early convalescent phase and convalescent phase of 34 EBV-IM children and compared with that of 24 healthy donors. RESULTS: (1) The levels of CD23(+) and CD23(+)CD19(+) cells decreased and CD95(+), CD95(+)CD23(+), Bcl-2(+) cells increased markedly in IM patients in acute phase [CD95(+) cells (19.43 +/- 8.46)%; CD95(+)CD23(+) cells (1.81 +/- 1.71)%; Bcl-2(+) cells (23.41 +/- 26.47)%] and early convalescent phase [CD95(+) cells (12.94 +/- 5.05)%; CD95(+)CD23(+) (1.05 +/- 1.20)%; Bcl-2(+) cells (10.54 +/- 9.68)%], as compared with those of healthy controls [CD95(+) cells (10.39 +/- 2.90)%; CD95(+)CD23(+) cells (0.50 +/- 0.46)%; Bcl-2(+) cells (7.25 +/- 2.88)%]. The earlier the course of IM, the more abnormal the expressive levels. All the abnormal results returned to normal in convalescent phase. (2) Positive relationships were observed between the expressions of CD95(+)CD23(+) cells and that of CD23(+) cells, CD23(+)CD19(+) cells during acute and early convalescent phase, the expressions of Bcl-2(+), CD3(+) cells and CD23(+), CD23(+)CD19(+) cells during acute phase, the expressions of CD95(+)CD23(+) cells and Bcl-2(+) cells during acute phase, and the expressions of CD95(+)CD23(+) cells and CD95(+) cells during convalescent phase. CONCLUSION: The results indicate that CD95L-CD95 mediated apoptosis plays an important role in eliminating EBV-immortalized cells, which is counteracted partly by Bcl-2.

Antigens, CD19↗

Acyclovir treatment in infectious mononucleosis: a clinical and virological study.

Fifty-six patients with a clinical and laboratory diagnosis of infectious mononucleosis who had not been ill for more than seven days, were randomised for peroral treatment with acyclovir (800 mg five times daily) or placebo for seven days in a double blind trial. Clinical, virological and immunological parameters were monitored in each patient for six months. During treatment, shedding of Epstein-Barr virus' as assessed in 36 patients, was significantly reduced (p less than 0.001). However, virus production in the oropharynx returned to pre-treatment levels one week after the cessation of therapy. Virus was detected in 35 patients at enrollment and in 28 of 36 patients at the six-month control. No effect on the clinical course of the disease was noticed. The virus-specific antibody response was also unaffected. A significant reduction in spontaneous outgrowth of in vivo Epstein-Barr virus-infected B-lymphocytes was found at 180 days after treatment in four acyclovir-treated patients compared to six controls (p less than 0.001). In another three patients with over-whelming clinical symptoms causing airway obstruction and/or disseminated intravascular coagulopathy, treatment with intravenous acyclovir (10 mg/kg three times daily) was combined with prednisolone (0.7 mg/kg daily) for ten days. Virus shedding ceased transiently during treatment, but returned to initial levels within one week. A dramatic clinical effect on the pharyngeal oedema and general health of the two patients with airway obstruction was noticed, but was much less evident in a patient with intravascular coagulopathy.

Acyclovir↗

T lymphocyte anergy during acute infectious mononucleosis is restricted to the clonotypic receptor activation pathway.

The transient T cell anergy associated with acute infectious mononucleosis (IM) caused by the Epstein-Barr virus has been analysed in a sample of 14 IM children. Peripheral blood mononuclear cells (PBMC) obtained from IM patients showed a significant specific impairment in their proliferative response to both phytohaemagglutinin (PHA; P less than 0.05) and to an anti-CD3 MoAb (P less than 0.001), although both responses reached normal control levels by addition of a submitogenic dose of either phorbol myristate acetate (PMA) or recombinant IL-2 (rIL-2). In contrast, activation signals delivered through other surface molecules (CD2, CD28) or other transmembrane pathways (PMA plus a calcium ionophore) elicited normal or high proliferative responses in most IM PBMC. In a group of five patients tested, the synthesis of IL-2 by IM PBMC in the presence of PMA was impaired when PHA or anti-CD3 was used as stimulus, but it reached normal levels with anti-CD2 or ionophore. Lastly, PHA failed to induce IL-2 alpha receptor (IL-2R alpha) expression in IM PBMC from four tested patients, but the presence of PMA completely corrected this defect. Taken together, these results strongly suggest that the T cell anergy associated with acute IM is due to a T cell receptor (TCR)-specific impairment in the induction of genes involved in T cell proliferation (including those coding for IL-2 and IL-2R alpha) upon membrane signalling to otherwise normal T lymphocytes, since CD2, CD28 and certain transmembrane activation pathways are uncoupled from CD3 in these particular pathological conditions (and perhaps in most in vivo situations). This and other similar experimental approaches to transient secondary immunodeficiencies may help to unravel the physiopathological role of different surface molecules in T cell activation.

Acute Disease↗

Acute infectious mononucleosis stimulates the selective expression/expansion of V beta 6.1-3 and V beta 7 T cells.

Acute infectious mononucleosis (AIM) is caused by the Epstein-Barr virus (EBV) and is characterized by a proliferation of atypical lymphocytes, predominantly CD8+ T cells. Various diseases associated with T-cell activation have been shown to stimulate the selective expansion of certain V beta (variable region of the T-cell receptor beta chain) expressing T-cell populations. The purpose of this investigation was to determine if the proliferation of T cells accompanying AIM is associated with selective expression/expansion of distinct populations of V beta T cells. We determined V beta expression in eight patients with clinical and laboratory evidence of AIM, including an atypical lymphocytosis. Gel electrophoresis and quantitative analysis were performed on cDNA amplified by the polymerase chain reaction (PCR) using different V beta region primers. Gel electrophoresis analysis showed prominent V beta 6.1-3 and V beta 7 bands in all eight patients with AIM but not in the controls. Quantitative PCR analysis showed that the V beta 6.1-3 and V beta 7 mean PCR ratios increased, respectively, from 163.0 +/- 22.5 and 142.3 +/- 5.5 in controls to 339.9 +/- 38.8 (P < .03) and 396.1 +/- 45.6 (P < .01) in the eight patients with AIM. Two of the eight patients who had increased V beta 6.1-3 and V beta 7 expression were retested after clinical resolution of AIM and no longer had evidence of increased V beta 6.1-3 and V beta 7 T-cell expression. AIM is associated with a selective increased expression of V beta 6.1-3 and V beta 7 T cells present at the time of initial clinical symptoms and atypical lymphocytosis. This increased expression resolves following recovery from AIM. This V beta-specific selective expression resembles the super-antigen response seen after staphylococcal toxin stimulation and may be caused by EBV triggering of selective expansion of V beta 6.1-3 and V beta 7 T-cell subsets.

Acute Disease↗

Positive serological tests for infectious mononucleosis in a patient with Mediterranean spotted fever.

The Monospot test is an accurate and reliable test for infectious mononucleosis. False positive reactions have been seldom recorded. We report the conversion of the Monospot test from negative to positive during the course of Mediterranean spotted fever. Changes in the titre of the Paul-Bunnell test were observed also. The importance of measuring IgM antibodies as part of the routine investigation of infectious diseases is emphasised.

Adult↗