Hormone therapy affecting interferon defense in children with infectious mononucleosis

23 children diagnosed with acute infectious mononucleosis were hospitalized and examined after a short prednisolone treatment course. Related interferon status during infection was compared with that in 38 patients with acute infectious mononucleosis receiving no hormone therapy. Interferon status was investigated by Ershov method, allowing to estimate amount of interferon in the blood serum samples or patient blood cell culture by assessing interferon biological activity. Along with measuring IFNα or IFNγ biological activity, their level was quantified by using enzyme immunoassay. Immunological examination conducted on the next day after the end of hormone therapy revealed sharply decreased potential of patient blood cells to produce both IFNα and IFNγ. The multiplicity of IFNα and IFNγ titer reduction in various patients varied by 4–5 and 3–4-fold, respectively. The concentration of IFNα, determined by ELISA, decreased by 4–6-fold, whereas for IFNγ — by 1.5–2-fold. A follow-up examination 1 month after discharge from the clinic showed that mean IFNα titer in children aged 3–6 years and treated with prednisolone was significantly reduced compared to the baseline, whereas most patients receiving no hormone therapy had normal IFNα production. The change in the level of IFNα 1 month after hormone therapy in 7–14-year age group was similar. IFNγ production quickly recovered, and 1 month after discharge from the clinic, its concentration in culture supernatants from patients reached 10–15 ng/ml, exceeding normal values more than twice. The biological activity of IFNγ in these culture supernatants was significantly higher than those immediately after hormone therapy, whereas in 3–6-year-old group of patients it was also higher than baseline level. These results can serve as a laboratory justification for including recombinant IFNα-2b drugs in the therapy of such patients, presumably immediately after the end of hormone course. Overall, laboratory justified administration of interferon preparations seems to be necessary to determine optimal timepoint for applying such drugs to increase effectiveness for achieving a durable patient recovery.

HLA-DR Expression Level in CD8+ T Cells Correlates With the Severity of Children With Acute Infectious Mononucleosis

BackgroundThis study aimed to assess the host immune signatures associated with EBV infection and its clinical value in indicating the severity of children with acute infectious mononucleosis (IM).MethodsTwenty-eight pediatric patients with IM aged 3–8 years were enrolled. The immune phenotypes and cytokine secretion capability of T cells were detected.ResultsThe percentages and absolute numbers of CD3+ and CD8+ T cells were significantly increased in IM patients compared with HCs. The percentages of Naïve CD4+ and CD8+ T cells were decreased but with increased percentages of memory CD4+ and CD8+ T subsets. Our results showed the upregulation of active marker HLA-DR, TCR-αβ, and inhibitory receptors PD-1, TIGIT in CD8+ T cells from IM patients, which suggested that effective cytotoxic T cells were highly against EBV infection. However, EBV exposure impaired the cytokine (IFN-γ, IL-2, and TNF-α) secretion capability of CD4+ and CD8+ T cells after stimulation with PMA/ionomycin in vitro. Multivariate analysis revealed that the percentage of HLA-DR+ CD8+ T cells was an independent prognostic marker for IM. The percentage of HLA-DR+ CD8+ T cells was significantly correlated with high viral load and abnormal liver function results.ConclusionRobust expansion and upregulation of HLA-DR in CD8+ T cells, accompanied with impaired cytokine secretion, were typical characteristics of children with acute IM. The percentage of HLA-DR+ CD8+ T cells might be used as a prominent marker not only for the early diagnosis but also for indicating the severity of IM.

Methodological bases of differential detection of the EBV1/EBV2 and HHV6A/HHV6B

In Russia, of the whole variety of EBV- and HHV6-etiology diseases, only infectious mononucleosis is subject to official statistical reporting, which significantly complicates an objective assessment of their etiological structure, incidence rate, characteristics of the development of the epidemic process. Currently, information on the genetic heterogeneity of EBV, even at the level of the main types (EBV1 and EBV2), as well as HHV6A and HHV6B, their prevalence and clinical significance are limited mainly by foreign data. In Russia, there are isolated publications devoted to this issue. At the same time, the study of the circulation of genetic types (variants) and the use of this information in the implementation of epidemiological surveillance of some other infections have already become routine practice. One of the key issues is the level of development of laboratory support for molecular genetic monitoring. The purpose of this work was to improve the methodological base of differential detection of HHV6A/B and the main types of EBV. The material for the study was peripheral blood leukocytes of children aged 1-15 years with acute infectious mononucleosis (n = 50) and without clinical symptoms of this disease (n = 29). The detection and quantification of EBV DNA and HHV6 DNA was performed using real-time PCR. For differential determination of EBV1/EBV2 and HHV6A /HHV6B, an optimized one-round PCR with electrophoretic detection of amplification products in an agarose gel was used. According to the results of our own research, the frequency of detection of EBV DNA and HHV6 DNA in acute infectious mononucleosis was 74% and 72%, and in the control group - 35% and 74%, respectively. It was found that among the examined children of the Nizhny Novgorod region, EBV1 and HHV6B prevail in the viral population, which is consistent with existing views about their geographical distribution in the adjacent territories. EBV2 was found in a single sample in the control group only. HHV6A was not detected in any of the studied groups. The methodological approach optimized in this work makes it possible to separately detect HHV6A/HHV6B and the main types of EBV according to a single laboratory protocol, and in combination with an additional stage of DNA concentration increases the diagnostic sensitivity of PCR analysis, minimizes the proportion of discordant and false negative results. Such an integrated approach can be applied for diagnostic, epidemiological and research purposes.

Epstein-Barr virus uveitis after intravitreal triamcinolone injection

Purpose: To report a case of Epstein-Barr virus (EBV) uveitis after intravitreal triamcinolone injection. Methods: Observational case report. Results: A 66-year-old male presented with bilateral intermediate uveitis, left macular branch retinal vein occlusion, and left macular edema 3 months following acute infectious mononucleosis. He received systemic prednisolone, methotrexate, and intravitreal bevacizumab with partial response. Intravitreal triamcinolone was given for recurrent macular edema, which led to the development of severe panuveitis with positive EBV PCR in aqueous humour. This was successfully treated with high-dose systemic valaciclovir and topical prednisolone. Conclusion: Non-infectious uveitis may become infectious following intravitreal steroid administration triggering intraocular viral replication. Intraocular fluid should be tested in cases which are suspicious for infection and EBV should be considered a differential diagnosis, particularly if PCR is negative for more common viral etiologies.

Cytokine production and clinical and laboratory aspects of EBV-associated acute infectious mononucleosis in children

Objective. To analyze clinical and laboratory parameters, as well as the dynamics of cytokine production in children of different ages with acute infectious mononucleosis caused by Epstein-Barr virus (EBV mononucleosis). Patients and methods. We examined two groups of patients: group I included 20 children aged 1 to 7 years, whereas group II included 29 children aged 8 to 17 years. All study participants were tested in the acute phase of the disease and in early convalescence. We evaluated serum levels of interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α), and interferon-α (IFN-α) using enzyme-linked immunosorbent assay (ELISA) (standard Vektor-Best kits, Russia). Data analysis was performed using Microsoft Excel 2019 for Windows and IBM SPSS statistics; we applied the methods of non-parametric statistics. Differences were considered significant at p < 0.05. Results. The majority of children had fever, intoxication, acute tonsillitis, and enlarged cervical lymph nodes. Laboratory makers, such as lymphocytosis, neutropenia, and thrombocytopenia were more pronounced in children from group II. In both groups, the level of cytokines in the acute period of the disease was higher than the discriminatory one. In early convalescence, patients from group I demonstrated more significant reduction in the cytokine level than patients from group II (р < 0.05). In children over 7 years of age, the levels of IL-10 and TNF-α positively correlated with the disease duration (p < 0.01 and p < 0.05, respectively) Conclusion. The level of cytokine production in acute EBV mononucleosis depends on patients’ age. Concentrations of IL-10 and TNF-α can serve as markers reflecting the severity of EBV mononucleosis and can be used for disease prognosis. Key words: EBV mononucleosis, children, cytokines, IL-6, IL-10, TNF-α, IFN-α

The vagaries of IgM: a case report of EBV infection with concomitantly false-positive IgM for CMV, VZV, and HSV

Background Serum IgM (immunoglobulin M) testing is commonly used to diagnose acute viral infections. However, most clinicians are unaware of the vagaries of IgM testing, including antigenic cross-reactivity between multiple viruses and risk misdiagnosis. Case presentation We report a case of infectious mononucleosis with concomitantly positive IgM for EBV, CMV, VZV, and HSV. A 26-year-old man presented with acute infectious mononucleosis picture. His blood work showed a total bilirubin level of 7.7 mg/dl, ALT 1077 U/L, AST 806 U/L, ALP 325 U/L, and INR 1.0. Monospot was positive; peripheral blood smear showed atypical lymphocytes; however, because EBV infectious mononucleosis does not typically cause elevation of liver enzymes over 1000, other etiologies were explored. Tests for hepatitis A, B, C, HIV, ANA, and ASMA returned negative. IgM for EBV-VCA, CMV, HSV, and VZV all returned positive, and the diagnosis of EBV IM was called into question. Subsequent tests of CMV and HSV PCR for viral load were negative (VZV was not clinically suspected), and later on, EBV-EBNA returned negative and EBV-VCA IgM and IgG returned positive, confirming the diagnosis of acute EBV infection. Conclusion We believe that IgM seropositivity can result from cross-reactivity among several viruses (especially herpes viruses), and although often relied on, a positive IgM should not serve as the sole determinant for diagnosis of acute viral infections.

Evaluation of Epstein-Barr Virus Indirect Immunofluorescence Assay Results

Background: The aim of this study was to diagnose Epstein-Barr virüs (EBV) infection using indirect immunofluorescence assay (IIFA) method and to evaluate these patients clinically. More than 90% of children are infected with EBV until the age of six. Methods: The tests were studied by using EBV IIFA method. A total of 247 patients, 186 adults, and 61 children, were included in the study. Results: 5 (2.7%) of the adults were EBV-Capsid antigen (EBV-CA) IgM positive, 175 (94.1%) were IgG positive and 6 (3.2%) were seronegative. 10 (16.4%) of the child patients were IgM positive, which is considered as an acute infectious mononucleosis (IM) infection marker, whereas the child patients had a significantly higher IgM rate than adults (p <0.001). 39 (63.9%) of the child patients were IgG positive and 12 (19.7%) of them were seronegative. The rate of IgG positivity in children was significantly lower than in adults (p <0.001).There was no significant difference between the genders in terms of IgM and IgG positivity rates in both adults and children(p> 0.05 for each).Conclusion: These results suggest that the presence of an acute EBV infection should be considered when the patient has viral diseases with similar clinical picture especially for children.

A typical Presentation of Acute Infectious Mononucleosis (AIM) with Isolated Hyperbilirubinemia

Epstein - Barr virus (EBV) induced hepatitis and subsequent hyperbilirubinemia is a strikingly rare cause of jaundice. Lack of other common infectious mononucleosis symptoms makes the diagnosis difficult with history and physical exam alone. With differential diagnoses more commonly including HAV, HCV, and HBV hepatitis infections; alcoholic hepatitis; autoimmune hepatitis; and hepatocellular carcinoma, suspicion for EBV induced hepatitis is often low. We present a noteworthy case of isolated hyperbilirubinemia due to EBV virus confirmed with biopsy, without other infectious mononucleosis symptoms such as fever, sore throat, or splenomegaly. Furthermore, we review the pathophysiology, diagnosis, treatment, and prognosis of EBV-induced hepatitis.

EBV-requisitioning physicians’ guess on fatigue state 6 months after acute EBV infection

We assessed referring medical practitioner’s ability to predict chronic fatigue development in adolescents presenting with acute infectious mononucleosis. Compared with ‘not fatigued’ being predicted as ‘unsurely fatigued’ and ‘likely fatigued’ were both strongly associated with developing fatigue 6 months later (OR 2.5, 95% CI 1.16% to 5.47% and 3.2, 95% CI 1.19% to 8.61%, respectively, P=0.012). The positive and negative predictive values were 66% and 62%, respectively. Disentangling the physician’s intuition may be of interest in further investigations of risk factors and prophylactic factors for fatigue development.