Epstein-Barr Virus Viral Load Research Articles

Effect of Epstein-Barr virus on macrophage M2/M1 migration and EphA2 expression in adverse drug reactions.

This study aims to investigate the effect of Epstein-Barr virus (EBV) reactivation or EBV reactivation with dexamethasone (DXM) in patients with adverse drug reactions (ADRs) through evaluating the levels of monocyte, macrophage M2/M1, and cytokines, and investigating whether expression of EBV receptor EphA2 could specifically influence EBV activation in ADRs. We performed a prospective longitudinal study to analyze the monocytes, macrophages, M2/M1 ratio, and cytokines, including interleukin (IL)-4, IL-13, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, IFN-β, C-X-C motif chemokine ligand (CXCL)9, and CXCL10, in patients with maculopapular exanthema (MPE) and Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN), and control groups after disease onset. Skin biopsy samples from these patients were subjected to hematoxylin and eosin (H&E) staining to examine tissue architecture and inflammatory cell infiltration, as well as Epstein-Barr virus-encoded RNA (EBER) staining to detect the presence of EBV within the skin lesions. Peripheral blood mononuclear cells collected from these patients were co-cultured with EBV or EBV combined with DXM to assess the impact on monocytes, macrophages, the M2/M1 ratio, and the associated cytokine profile. Furthermore, we sought to identify which cytokines might be crucial in mediating the interaction between the M2/M1 ratio and EBV. EPhA2 expression was evaluated to determine its role in the reactivation of EBV and its correlation with increased viral load in MPE and SJS/TEN patients. Selective depletion of macrophages occurred during the acute stage of SJS/TEN, while a shift towards M2 macrophages was observed in MPE. Both IFN-β and CXCL9 levels were elevated in MPE and SJS/TEN. Additionally, our study demonstrated the presence of EBV in the skin lesions of SJS/TEN and MPE patients through H&E and EBER staining, confirming EBV's involvement in these conditions. Activation of EBV and EBV combined with DXM led to a shift from M1 to M2 macrophages, accompanied by increased levels of IL-4, IFN-γ, and CXCL9 in MPE and SJS/TEN, compared to healthy individuals. Specifically, EBV combined with DXM primarily drove IFN-γ and IL-4 expansions in MPE, while CXCL9 was predominantly elevated in SJS/TEN. The increased IL-4 levels were associated with the relative rise in EBV viral loads after EBV combined with DXM stimulation. Furthermore, EphA2 expression in monocytes was significantly higher in SJS/TEN and MPE patients compared to controls, with further increases on EBV stimulation. This elevation in EPhA2 correlated with increased EBV viral load, particularly in MPE and SJS/TEN patients. The gradual shift from M1 to M2 cell development observed during the clinical course of MPE and SJS/TEN is mediated by the predominance of EBV and EBV with DXM at the acute stage, leading to elevated IL-4, IFN-γ, and CXCL9 levels, which may exacerbate allergic reactions. The elevation in EPhA2 correlated with increased EBV viral load, particularly in MPE and SJS/TEN patients, suggesting that adverse drug reactions may induce EPhA2 expression, facilitating EBV replication and activation. EphA2 could thus serve as an indicator of EBV activation and a marker for assessing the risk of EBV in patients with adverse drug reactions.

Does Epstein-Barr virus and intracellular Toll-like receptors affect the course of Hashimoto's disease? Findings from studies on newly diagnosed patients.

Epstein-Barr virus (EBV) reactivation is increasingly recognized as a potential exacerbator of autoimmune diseases, including Hashimoto's thyroiditis (HT). This study examined the association between EBV reactivation and intracellular Toll-like receptors (TLRs) expression in newly diagnosed, untreated HT patients. The aim was to determine whether EBV reactivation and specific TLRs (TLR3, TLR7, TLR8, and TLR9) contribute to the pathogenesis and progression of HT. A cohort was 54 newly diagnosed, untreated patients with HT and 20 healthy volunteers (HV). Also, the HV were similarin age and gender. Blood samples were collected to assess EBV viral load and intracellular expression levels of TLR3, TLR7, TLR8, and TLR9 using flow cytometry. Specific anti-EBV antibodies (e.g., VCA IgM, VCA IgG, EBNA-1 IgM, EBNA-1 IgG) were measured to identify signs of EBV reactivation, while soluble TLRs (sTLR3, sTLR7, sTLR8, and sTLR9) were quantified in serum using ELISA. Notably, this study's patients with Hashimoto's thyroiditiswere not euthyroid, as they exhibited significantly lower TSH levels and elevated fT3 and fT4 levels compared to healthy controls. Results showed a significant increase in EBV reactivation among HT patients, with 26 of 54 (48.1%) testing positive for EBV DNA, compared to none in the control group. HT patients with reactivated EBV showed significantly higher levels of intracellular expression of TLR3, TLR7, and TLR9, with TLR3+ cells constituting an average of 4.72% of CD4+ lymphocytes compared to 0.69% in the control group, suggesting a potential synergistic effect. In addition, soluble TLR levels were increased in HT patients with reactivated EBV, suggesting a potential role in potentiating autoimmune responses. These findings highlight the importance of considering both viral reactivation and TLR activity in the treatment of HT. Understanding the interplay between EBV and intracellular TLRs may lead to the development of new therapeutic approaches to mitigate the impact of these factors on disease progression. Further research is warranted to investigate the mechanisms underlying this interaction and its implications for treatment strategies.

P35 Pyrexia of unknown origin in a patient with long standing rheumatoid arthritis

Abstract Introduction Pyrexia of unknown origin (PUO) can be challenging in treated rheumatic auto-immune disease. Many conditions may have fever as part of the symptom complex, treatment may mask signs and symptoms and can also make opportunistic infection more likely. Unusual or fastidious infections can be particularly difficult to diagnose. We describe a case of ureaplasma infection that evaded detection for a prolonged period in a female with long standing rheumatoid arthritis (RA). Case description 47 year old female presented January 2023 with fevers, weight loss and persistently high CRP. RA diagnosed in 1998. Erosive wrist and MTP disease, initial RF 32, CCP 8. Treatment- 5-10 mg prednisolone since 1998, adalimumab 2010, rituximab 2013-2022. Normal Ig levels, IgG lamda paraprotein since 2016 (too low to quantify). Frequency and dysuria March 2022. Cystoscopy- small bladder; biopsies in May 2022 and August 2023 showed acute on chronic inflammatory infiltrate with granulation tissue. Interstitial cystitis diagnosed. She had recurrent episodes of pyuria and was given several courses of antibiotics, including meropenem, temocillin, fluconazole, gentamicin. Despite the recurrent pyuria no organisms were cultured. CT CAP February 2023 -ground glass opacity right lower lobe. Later CXR clear. Quantiferon negative, also negative 2016, 2019. Other normal / negative tests- TTG, HIV, Hep ABCE, TFTs, EBV viral load. EBV, CMV IgG +ve. CMV titres 696 c/ml June 2023. CD 19 undetectable. IgG4 0.02 g/L. MPO, PR3 negative. BK virus was detected in urine. There was no sustained response to iv methylprednisolone and a course of filgotinib given for possible systemic RA symptoms. She developed a dry cough and shortness of breath. PET-CT July 2023 - bilateral FDG avid large confluent areas of consolidation. Urine + bronchial washings were negative for TB growth at 42 days. However, in view of deterioration empirical TB treatment was started July 2023. CT urogram in September 2023 showed likely bilateral renal abcesses, as well as bilateral hydronephrosis. Left renal cyst aspirate was performed and empirical doxycycline started at 100 mg bd. Ureaplasma detected from the aspirate on PCR. 3 weeks doxycyline given with rapid resolution of fevers and CRP. February 2024- cystectomy and ileal conduit in for refractory interstitial cystitis with a poorly compliant, small capacity, thick walled fibrotic bladder which had caused bilateral ureteric obstruction. She has made a good recovery. TB treatment course was completed. Discussion This case shows the difficulties in diagnosing infection from a little known and difficult to detect organism, ureaplasma. Discovered in the 1950s, it has no cell wall and very small genome and is therefore an obligate parasite /saprophyte. Together with mycoplasma it belongs to the bacterial class of Mollicutes. It cannot be routinely cultured nor gram stained. PCR or specialist culture is required. It may be present as a commensal but has been implicated in symptomatic genitourinary infections. It has also been implicated in native and non-native joint infections, and in infants can cause a pneumonic illness. Empirical treatment may be indicated due to long turn around times for diagnosis. There is some evidence that reduced humoral immunity may be a contributory factor to infection. We have found a previous case described of ureaplasma renal abcesses in a patient with MS and neurogenic bladder treated with rituximab. Our patient had normal immunoglobulin levels though still had profound CD19 suppression. Further immune suppression resulted from methylprednisolone and course of filgotinib, which may have accelerated the course of infection. Ureaplasma and mycoplasma do not have a cell wall, thus making beta-lactam antibiotics and vancomycin ineffective. Cyclines, josamycin and fluroquinolones are effective, though there is increasing resistance developing. We gave a relatively long course of doxycycline as our patient was very unwell and had apparent extensive urinary tract involvement. BK virus seropositivity is present in 90% of the normal population and asymptomatic viruria in up to 20%. However, it is a cause of renal tract inflammation in the immunosuppressed – in particular post-transplant. We felt it was unlikely to be significant in our patient in view of the clinical picture, lack of BK viraemia and negative viral findings on bladder biopsy. Key learning points • Ureaplasma should be on the list of organisms to consider in cases of PUO when the more common infective causes have been ruled out, particularly in immune suppressed patients. Patience and close collaboration with colleagues is essential. This patient had multiple MDT meetings involving infectious disease, respiratory, secondary and tertiary care urology and rheumatology specialists. The collaboration was invaluable. A course of doxycycline may be considered as a relatively low risk approach while awaiting microbiological confirmation in such cases.

Human β-Defensin 2 as a Link between EBV Infection and Multiple Sclerosis

Multiple sclerosis (MS) is the result of a complex interaction between environmental factors and genetic predisposition. Epstein-Barr virus (EBV) is a significant environmental risk factor associated with MS. Human- beta defensin 2 (HβD-2) is a crucial innate immune response proteint that as an inflammatory marker protects humans from pathogens invading the body like viruses. Therefore, a case-control study was conducted on 90 subjected (48 patients with MS and 42 healthy controls) to examine the role of HβD-2 in EBV viral load infection and MS patients. An enzyme-linked immunosorbentassay kit was used to measure HβD-2 levels in serum. The viral load of EBV was determined using a real-time PCR. The findings revealed that median HβD-2 levels in patients were significantly lower than in controls. (96.62 vs.124.8 ng/mL; p < 0.01). According to the receiver operating characteristic curve analysis, HβD-2 is good predictor for MS disease (area under the curve=0.739; p < 0.001). EBV frequency was lower in MS patients compared to the controls, although difference was not statistically significant (25 vs. 38.1%; p = 0.181).On other hand, the median (EBV) load was substantially greater in patients than in healthy controls ( 8.55 vs. 1.4 DNA copy/100 cells). This study concluded that HβD-2 levels showed no significant differences in each age group, sex, Expanded Disability Status Scale (EDSS) of multiple sclerosis patients, rather showed an inverse significant correlation (correlation coefficient = - 0.432) with EBV load, which protect against human viral infections.

Epstein-Barr virus infection induces tissue-resident memory T cells in mucosal lymphoid tissues.

EBV contributes to around 2% of all tumors worldwide. Simultaneously, more than 90% of healthy human adults persistently carry EBV without clinical symptoms. In most EBV carriers, it is thought that virus-induced tumorigenesis is prevented by cell-mediated immunity. Specifically, memory CD8+ T cells recognize EBV-infected cells during latent and lytic infection. Using a symptomatic primary infection model, similar to infectious mononucleosis (IM), we found EBV-induced CD8+ tissue resident memory T cells (TRMs) in mice with a humanized immune system. These human TRMs were preferentially established after intranasal EBV infection in nasal-associated lymphoid tissues (NALT), equivalent to tonsils, the primary site of EBV infection in humans. They expressed canonical TRM markers, including CD69, CD103, and BLIMP-1, as well as granzyme B, CD107a, and CCL5. Despite cytotoxic activity and cytokine production ex vivo, these TRMs demonstrated reduced CD27 expression and proliferation and failed to control EBV viral loads in the NALT during infection, although effector memory T cells (TEMs) controlled viral titers in spleen and blood. Overall, TRMs are established in mucosal lymphoid tissues by EBV infection, but primarily, systemic CD8+ T cell expansion seems to control viral loads in the context of IM-like infection.

Exploring atypical manifestations and multisystem involvement of Epstein-Barr virus infection in hospitalized pediatric patients from Mexico: insights from a tertiary hospital (2012-2022).

Epstein-Barr virus (EBV) infection, with a global prevalence exceeding 95%, typically manifests in children as infectious mononucleosis. However, clinical practice frequently encounters diverse atypical presentations characterized by multisystem involvement, often resulting in an unfavorable clinical course. Our objective is to describe the clinical manifestations and results of EBV infection in a tertiary pediatric hospital in Mexico. An observational, transversal, retrospective, and descriptive study that included a systematic review of medical records (2012-2022) of patients under 18 years of age with detectable EBV particles in peripheral blood. The study included 26 patients with a median age of 5 years and a male predominance of 53.8%. Predominant symptoms were fever (85%) and lymphadenopathy (35%). Sixty-five percent had severe and atypical manifestations, including pneumonia and hepatic, hematologic-oncologic, and autoimmune diseases. Anemia, thrombocytopenia and leukopenia were common, with lymphocytosis in 19% of cases. The median EBV viral load was 2816 copies/mL (range: 555-355,500 copies/mL). Four deaths related to EBV infection were reported. Viral load in these cases also varied widely from 594 to 121,000 copies/mL. Supportive care was administered to 85% of patients, while others received antiviral treatment, steroids, and rituximab. Atypical manifestations were common, especially in children with multisystem involvement. EBV should be considered as a potential contributor to a diverse spectrum of clinical presentations, emphasizing the need for comprehensive evaluation and awareness in clinical diagnosis.

Emapalumab as a therapeutic intervention for Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis: A case series.

Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH) is characterized by a severe cytokine storm, heightened inflammatory response, and immune-mediated damage to tissues and organs. Standard treatment protocols for hemophagocytic lymphohistiocytosis often fall short in effectively controlling EBV-HLH, leading to a need for novel therapeutic options. Emapalumab, a monoclonal antibody targeting interferon-gamma, has shown promise due to its targeted cytokine modulation capabilities and favorable safety profile. This study aimed to evaluate the efficacy and safety of emapalumab in pediatric patients with EBV-HLH. The case series involved 4 pediatric patients diagnosed with EBV-HLH who did not achieve disease control despite receiving comprehensive treatment. All 4 pediatric patients were diagnosed with EBV-HLH. Emapalumab was introduced as an adjunctive therapeutic intervention alongside the HLH-94 or L-DEP regimens for these patients. Among the 4 patients, 1 experienced severe multiorgan dysfunction and opted to discontinue therapy. The remaining 3 patients showed controlled disease progression with significant clinical improvements following emapalumab administration. These improvements included reduced levels of inflammatory markers, normalization of blood counts and liver function, and decreased Epstein-Barr virus viral load. The findings suggest that emapalumab may be an effective and safe treatment option for pediatric EBV-HLH. However, further research is necessary to confirm these outcomes, especially in critically ill patients.

Splenic cyst deroofing complicated with B lymphoma

BackgroundSplenic cysts are uncommon and very rarely malignant therefore their treatment isn’t standardized. In case of symptomatic cysts different surgical approaches have been suggested. Primary malignant lymphoma of the spleen comprises less than 1% of non-Hodgkin’s lymphomas. To our knowledge, only 203 cases of splenic large B-cell lymphoma (LBCL) have been reported to date and only 2 of them were fibrin-associated splenic cysts.Case presentation27-year-old model with a 19 × 13 cm splenic cyst without data of malignancy in the preliminary study and therefore treated with laparoscopic deroofing. After histological diagnosis of LBCL with a fibrin/EBV-associated splenic pseudocyst, the patient received 4 cycles of Rituximab and a laparoscopic splenectomy was performed due to resurgence of the pseudocyst. No evidence of malignancy has been found during follow up (EBV viral load every 3 months during the first year, PET-CT every 6 months during the first year and annual afterwards) performed after the splenectomy.Discussion and conclusionsThe value of tumor markers and radiology for diagnosis of splenic cysts is put into question.Only 60 cases of Fibrin-associated LBCL (FA-LBCL) have been described in the literature therefore there are no treatment guidelines for them even though surgery together with systemic treatment has been the prevalent route with good results in most cases.

Quantitatively assessed blood leukocyte Epstein–Barr virus in adult HIV-infected patients

Coinfection with the human immunodeficiency virus (HIV) and Epstein–Barr virus (EBV) represents a current biomedical problem. The purpose of the study was to evaluate blood leukocyte EBV detection rate and viral load in adult HIV-infected patients. Materials and methods. There were examined blood leukocytes collected from 138 HIV(+) and 68 HIV(–) individuals aged 20–69 years. Statistical analysis was carried out differentiated according to the stages of HIV infection, the CD4+ T-lymphocyte count, and adherence to antiretroviral therapy. Results. It was shown that EBV DNA was detected significantly more often in HIV(+) vs HIV(–) individuals (70.3±3.9% and 48.5±6.1%, p = 0.008), with EBV viral load comprising 18 [5; 139] versus 2 [1; 3] copies/105 cells (р 0.001), respectively. It has been shown that the group of HIV(+) patients is heterogeneous in the frequency of EBV detection and viral load, with the peak EBV frequency (86.7±6.2%) and DNA (121 [34; 252] copies/105 cells) level observed in “naive” patients with severe immunodeficiency. Among “experienced” patients receiving therapy, the relative risk of detecting EBV DNA with low treatment adherence was significantly higher compared to those who developed high adherence (р 0.05). When the HIV viral load reached undetectable level, EBV DNA concentration was significantly lower where HIV RNA was detectable (1 [0; 8] versus 15 [1; 162] copies/105 cells, р 0.001). Detection of EBV DNA is associated with higher HIV viral load level and lower CD4+ T-lymphocyte count compared to patients with undetected EBV DNA. A relationship has been established between the CD4+ T-lymphocyte count in HIV(+) patients and the likelihood of active EBV infection. A threshold cut-off of 200 cells/μl was determined. CD4+ T-lymphocyte count 200 cells/μl vs ≥ 200 cells/μl (р 0.001) is associated with a 3.3-fold higher risk of detecting active EBV infection. Conclusion. It is necessary to continue interdisciplinary research to improve early diagnostics of EBV-associated diseases in HIV-infected individuals.

DRB1 locus alleles of HLA class II are associated with modulation of the immune response in different serological profiles of HIV-1/Epstein-Barr virus coinfection in the Brazilian Amazon region.

Epstein-Barr virus (EBV) infection involves distinct clinical and serological profiles. We evaluated the frequency of alleles of locus DRB1 of HLA class II in different serological profiles of EBV infection among HIV-1 infected patients. We recruited 19 patients with primary infection, 90 with serological transition and 467 with past infection by EBV, HIV-1 co-infection was 100% in primary infection and approximately 70% in other serological profiles. EBV viral load was quantified by real-time PCR, T lymphocyte quantification and cytokine level analysis were performed by flow cytometry, and HLA locus genotyping was performed by PCR-SSO. The DRB1*09 allele was associated with primary infection (p: 0.0477), and carriers of the allele showed changes in EBV viral load (p: 0.0485), CD8(+) T lymphocyte counts (p: 0.0206), double-positive T lymphocyte counts (p: 0.0093), IL-4 levels (p: 0.0464) and TNF levels (p: 0.0161). This allele was also frequent in HIV-coinfected individuals (p: 0.0023) and was related to the log10 HIV viral load (p: 0.0176) and CD8(+) T lymphocyte count (p: 0.0285). In primary infection, the log10 HIV viral load was high (p: 0.0060) and directly proportional to the EBV viral load (p: 0.0412). The DRB1*03 allele correlated with serological transition (p: 0.0477), EBV viral load (p: 0.0015), CD4(+) T lymphocyte count (p: 0.0112), CD8(+) T lymphocyte count (p: 0.0260), double-negative T lymphocyte count (p: 0.0540), IL-4 levels (p: 0.0478) and IL-6 levels (p: 0.0175). In the serological transition group, the log10 HIV viral load was high (p: 0.0060), but it was not associated with the EBV viral load (p: 0.1214). Past infection was related to the DRB1*16 allele (p: 0.0477), with carriers displaying IgG levels (p: 0.0020), CD4(+) T lymphocyte counts (p: 0.0116) and suggestive CD8(+) T count alterations (p: 0.0602). The DRB01*16 allele was also common in HIV-1 patients with past EBV infection (p: 0.0192); however, the allele was not associated with clinical markers of HIV-1 infection. Our results suggest that HLA class II alleles may be associated with the modulation of the serological profiles of the immune response to Epstein-Barr virus infection in patients coinfected with HIV-1.

Post-transplant lymphoproliferative disorder associated Epstein-Barr virus DNAemia after liver transplantation in children: Experience from single center.

The most prevalent malignancy that complicates both adult and pediatric solid organ transplantation is post-transplant lymphoproliferative disorder (PTLD). This study aimed to analyze the clinical and pathological characteristics, treatments, and outcomes of Epstein-Barr virus (EBV) DNAemia and PTLD in pediatric liver transplant recipients. A retrospective chart review was performed on 112 patients less than 18 years of age who underwent isolated orthotopic liver transplantation (OLT) between 2010 and 2022 at Ege University Children's Hospital. Data gathered for 1-year post-OLT included age at OLT, EBV, immunoglobulin (Ig)M/IgG status of the donor and recipient, indication for OLT, induction regimen, all immunosuppression levels, date and result of EBV polymerase chain reaction testing, rejection episodes documented by liver biopsy, and the development of PTLD. Forty-nine patients (43.75%) developed EBV DNAemia (median interval from surgery: 2 months, min-max: 2-36), of which 43 (87.8%) grafts came from living donors, and 6 (12.2%) came from deceased donors. Nine (18.4%) patients died during follow-up, and eight (16.3%) developed PTLD. Of these 8 patients; five patients developed EBV-related disease, one child developed hemophagocytic lymphohistiocytosis, one developed aplastic anemia, and one child developed B cell lymphoma. When PTLD patients and without-PTLD patients were compared, pediatric intensive care unit hospitalization, abnormal bone marrow biopsy findings, lymphadenopathy, age at diagnosis of EBV DNAemia, EBV viral load, tacrolimus (FK 506) pre-infection, were higher and tacrolimus 1-month levels were lower in patients with PTLD (p < 0.05). In logistic regression analysis, we showed that the age at diagnosis of EBV DNAemia was significantly higher in children with PTLD (p = 0.045; OR: 1.389; 95% CI: 1.007-1.914). PTLD is a rare but severe complication associated with EBV after OLT. This study demonstrated that PTLD is associated with older age, higher tacrolimus blood levels before EBV DNAemia, and higher peak EBV viral load at 1 month of EBV DNAemia.

Impact of Antiretroviral Therapy on the Progression of EBV-Associated Lymphomas

Objective — to determine the impact of antiretroviral therapy (ART) on the progression of lymphomas induced by Epstein—Barr virus (EBV) in patients infected with human immunodeficiency virus (HIV). Materials and methods. We analysed clinical-epidemiological, laboratory and serological indicators of 57 HIV-infected patients with clinically and laboratory-confirmed EBV co-infection. Patients were divided into two groups: the primary group consisted of 9 patients with lymphomas (66.7 % of whom received ART without concurrent chemotherapy with cytostatic antitumor agents, while the rest did not receive ART), and the control group included 48 patients without neoplasms. Results and discussion. To assess the effectiveness of ART in treating EBV-induced lymphomas, we analysed the general clinical, immunological and virological indicators of patients at the start of observation and after 6 months of therapy. In patients with EBV-associated lymphomas receiving ART, there was an increase in CD4+ lymphocytes, a decrease in HIV viral load, an increase in hemoglobin levels (p &lt; 0.05), and a significant reduction in EBV viral load (p &lt; 0.01). In patients with EBV-associated lymphomas who did not receive ART, there was a further decrease in CD4+ lymphocytes and an increase in EBV viral load. Two clinical cases are presented: in one, a patient received palliative chemotherapy according to the CHOP regimen but did not receive ART due to an absolute refusal because of poor drug tolerance; in the other case, the patient received ART without cytostatic antitumor drugs due to an absolute refusal. Conclusions. The study results established that the use of ART in patients with EBV-associated lymphomas not only improved the primary indicators characteristic of HIV but also significantly reduced the EBV viral load (p &lt; 0.01). The dynamics of the main immunological and virological indicators in patients with EBV-associated lymphomas who did not receive ART were negative, as evidenced by the further decrease in CD4+ lymphocytes and an increase in EBV viral load. From the clinical cases presented, it can be concluded that the timely administration of ART to patients with EBV-associated lymphomas is a crucial therapeutic tool that not only facilitates positive immunological and virological changes in HIV infection but also affects the quality and duration of life in patients with EBV-associated lymphomas.

Experience with Epstein barr virus and Cytomegalovirus infection in pediatric liver transplant recipients: A 2014–2017 study

IntroductionThis study presents our experience with Epstein Barr virus (EBV) and Cytomegalovirus (CMV) infections in the pediatric liver transplant population and their potential association with the development of the post-transplant lymphoproliferative syndrome (PTLD). Patients and methodsThis retrospective descriptive study covers the period from 2014 to 2017 and includes pediatric liver transplanted recipients who underwent viral load monitoring for EVB and CMV during the first-year post-transplant. ResultsA total of 89 patients were included in the study, with a median age of 0.68 years (RIQ 0.31–0.96) (10.8 months RIQ 8.4–25.2). The most common underlying pathology leading to transplantation was biliary atresia, observed in 55 (61.6 %) cases. Regarding EBV viral loads, values exceeding 10,000 copies/ml were observed in 9 (8 %) patients at 3 months, 33 (29.3 %) at 6 months, 31 (27.6 %) at 9 months and 25 (22.3 %) at 12 months post-transplantation. The probability of developing EBV infection within one-year post-transplantation was 81.3 %, while the probability of CMV infection was 29 %. A total of 8 (8.9 %) biopsy-confirmed graft rejections occurred, only 1 was EBV and CMV-negative. The likelihood of graft rejection in patients with EBV infection was 21.5 %, and for CMV it was 20.8 %. Importantly, only one case of PTLD was documented during 12 months follow-up. ConclusionCharacterizing this pediatric liver transplant population and monitoring EBV and CMV viral loads enables timely interventions, potentially reducing the risk of PTLD and graft rejection.

Disease-Modifying Treatments for Multiple Sclerosis Affect Measures of Cellular Immune Responses to EBNA-1 Peptides.

Epstein-Barr virus (EBV) has been strongly implicated in the pathogenesis of multiple sclerosis (MS). Despite this, there are no routinely used tests to measure cellular response to EBV. In this study, we analyzed the cellular response to EBV nuclear antigen-1 (EBNA-1) in people with MS (pwMS) using a whole blood assay. This cross-sectional study took place in a dedicated MS clinic in a university hospital. We recruited healthy controls, people with epilepsy (PWE), and pwMS taking a range of disease-modifying treatments (DMTs) including natalizumab, anti-CD20 monoclonal antibodies (mAbs), dimethyl fumarate (DMF), and also treatment naïve. Whole blood samples were stimulated with commercially available PepTivator EBNA1 peptides and a control virus-cytomegalovirus (CMV) peptide. We recorded the cellular response to stimulation with both interferon gamma (IFN-γ) and interleukin-2 (IL-2). We also compared the cellular responses to EBNA1 with IgG responses to EBNA1, viral capsid antigen (VCA), and EBV viral load. We recruited 86 pwMS, with relapsing remitting MS, in this group, and we observed a higher level of cellular response recorded with IFN-γ (0.79 IU/mL ± 1.36) vs healthy controls (0.29 IU/mL ± 0.90, p = 0.0048) and PWE (0.17 IU/mL ± 0.33, p = 0.0088). Treatment with either anti-CD20 mAbs (0.28 IU/mL ± 0.57) or DMF (0.07 IU/mL ± 0.15) resulted in a cellular response equivalent to control levels or in PWE (p = 0.26). The results of recording IL-2 response were concordant with IFN-γ: with suppression also seen with anti-CD20 mAbs and DMF. By contrast, we did not record any differential effect of DMTs on the levels of IgG to either EBNA-1 or VCA. Nor did we observe differences in cellular response to cytomegalovirus between groups. This study demonstrates how testing and recording the cellular response to EBNA-1 in pwMS may be beneficial. EBNA-1 stimulation of whole blood samples produced higher levels of IFN-γ and IL-2 in pwMS compared with controls and PWE. In addition, we show a differential effect of currently available DMTs on this response. The functional assay deployed uses whole blood samples with minimal preprocessing suggesting that employment as a treatment response measure in clinical trials targeting EBV may be possible.

Secondary hemophagocytic lymphohistiocytosis with associated central pontine myelinolysis

Abstract A 34-year-old previously healthy male presented to the emergency department with an altered mental state and unexplained intermittent fevers persisting for three weeks. Initially, his symptoms mimicked an ear infection, for which he received a prescription for Augmentin. As his condition deteriorated, additional symptoms emerged, including bilateral ear pain, fever, and diffuse headaches. Initial evaluations revealed acute hepatic encephalopathy, mastoiditis, and pancytopenia. Further tests uncovered hepatosplenomegaly, lymphadenopathy, and reactivated EBV. The patient met six of the eight HLH criteria, including fever above 38.3°C, cytopenia, elevated triglyceride and ferritin levels, and a bone marrow biopsy confirming hemophagocytosis. The treatment plan followed the HLH-2004 protocol, involving an eight-week induction therapy using etoposide, dexamethasone, and cyclosporine. Monitoring of disease-specific markers such as ferritin and IL-2R showed improvement. One month later, the patient experienced a throbbing headache, nausea, photophobia, and vision changes. MRI showed abnormal signals consistent with central pontine myelinolysis. Lumbar puncture findings included pleocytosis, elevated total protein, and a substantial EBV viral load, indicative of progressive CNS-HLH with persistent EBV reactivation. The patient is currently undergoing salvage therapy for EBV reactivation, with a poor prognosis for CNS involvement and allogeneic transplant preparations.

Epstein-Barr virus-associated post-transplant lymphoproliferative disorders in pediatric transplantation: A prospective multicenter study in the United States.

Epstein-Barr virus (EBV)-associated post-transplant lymphoproliferative disorders (PTLD) is the most common malignancy in children after transplant; however, difficulties for early detection may worsen the prognosis. The prospective, multicenter, study enrolled 944 children (≤21 years of age). Of these, 872 received liver, heart, kidney, intestinal, or multivisceral transplants in seven US centers between 2014 and 2019 (NCT02182986). In total, 34 pediatric EBV+ PTLD (3.9%) were identified by biopsy. Variables included sex, age, race, ethnicity, transplanted organ, EBV viral load, pre-transplant EBV serology, immunosuppression, response to chemotherapy and rituximab, and histopathological diagnosis. The uni-/multivariable competing risk analyses revealed the combination of EBV-seropositive donor and EBV-naïve recipient (D+R-) was a significant risk factor for PTLD development (sub-hazard ratio: 2.79 [1.34-5.78], p = .006) and EBV DNAemia (2.65 [1.72-4.09], p < .001). Patients with D+R- were significantly more associated with monomorphic/polymorphic PTLD than those with the other combinations (p = .02). Patients with monomorphic/polymorphic PTLD (n = 21) had significantly more EBV DNAemia than non-PTLD patients (p < .001) and an earlier clinical presentation of PTLD than patients with hyperplasias (p < .001), within 6-month post-transplant. Among non-liver transplant recipients, monomorphic/polymorphic PTLD were significantly more frequent than hyperplasias in patients ≥5 years of age at transplant (p = .01). D+R- is a risk factor for PTLD and EBV DNAemia and associated with the incidence of monomorphic/polymorphic PTLD. Intensive follow-up of EBV viral load within 6-month post-transplant, especially for patients with D+R- and/or non-liver transplant recipients ≥5 years of age at transplant, may help detect monomorphic/polymorphic PTLD early in pediatric transplant.

Analytical and clinical evaluation of the cobas Epstein–Barr virus test at a tertiary care cancer hospital

BackgroundEpstein–Barr Virus (EBV) viral loads in hematopoietic stem cell transplant (HSCT) recipients are typically monitored using quantitative molecular assays. The Cobas EBV test (Roche Molecular, Pleasanton, CA) has recently been FDA-cleared for the monitoring of EBV viral loads in plasma samples of transplant patients. In this study, we compared the viral loads obtained by a laboratory-developed test (EBV LDT) using Altona Analyte specific reagents (ASR) to those obtained on the Cobas EBV test. MethodsThe analytical performance of the assay was established using the EBV verification panel from Exact Diagnostics and the EBV ATCC strain B95-8. The clinical evaluation was performed using 343 plasma samples initially tested on the EBV LDT. ResultsThe analytical sensitivity (<18.8 IU/mL), precision (SD < 0.17 log) and linear range (35.0 IU/mL to 1E + 08 IU/mL) of the Cobas EBV assay established by the manufacturers were confirmed. The strength of the qualitative agreement was substantial between the cobas EBV and the EBV LDT (85.6 %; κ = 0.71) and almost perfect when discordant results were resolved (96.4 %; κ = 0.93). The quantitative agreement was moderate (82.9 %; κ = 0.53) with the viral load obtained on the Cobas EBV test being lower across the linear range of the two tests (mean log difference of 1.0). While the absolute values of the viral loads were markedly different, the overall trends observed in patients with multiple consecutive results were similar between the two tests. ConclusionsThe Cobas EBV test provides an accurate and valid, in vitro diagnostic (IVD) option for monitoring of EBV viral loads in transplant patients and should provide an opportunity for increased standardization and commutability of tests results across laboratories.

Analytical and Clinical Performance of the NeuMoDx™ Platform for Cytomegalovirus and Epstein-Barr Virus Viral Load Testing.

DNA assays for viral load (VL) monitoring are key tools in the management of immunocompromised patients with cytomegalovirus (CMV) or Epstein-Barr virus (EBV) infection. In this study, the analytical and clinical performances of the NeuMoDx™ CMV and EBV Quant Assays were compared with artus CMV and EBV QS-RGQ Kits in a primary hospital testing laboratory. Patient plasma samples previously tested using artus kits were randomly selected for testing by NeuMoDx assays. The NeuMoDx CMV Quant Assay and artus CMV QS-RGQ Kit limits of detection (LoDs) are 20.0 IU/mL and 69.7 IU/mL, respectively; 33/75 (44.0%) samples had CMV DNA levels above the LoD of both assays. The Pearson correlation coefficient was 0.9503; 20 samples (60.6%) had lower NeuMoDx CMV quantification values versus the artus kit. The LoD of the NeuMoDx EBV Quant Assay and artus EBV QS-RGQ Kit are 200 IU/mL and 22.29 IU/mL, respectively; 16/75 (21.3%) samples had EBV DNA levels above the LoD of both assays. The Pearson correlation coefficient was 0.8990. EBV quantification values with the NeuMoDx assay were higher versus the artus kit in 15 samples (93.8%). In conclusion, NeuMoDx CMV and EBV Quant Assays are sensitive and accurate tools for CMV and EBV DNA VL quantification.

EBV and multiple sclerosis: expression of LMP2A in MS patients.

Several evidences, including increased serum titers of Epstein-Barr virus (EBV)-specific antibodies and the presence of EBV DNA in brain of patients suggest a possible role of this virus in the pathogenesis of Multiple Sclerosis (MS), a chronic neurodegenerative disease with an unknown etiopathology. Aim of the present study is to verify if the expression of LMP2A and EBNA-1, two EBV genes, is altered in MS patients. EBV viral load, LMP2A and EBNA-1 gene expression and EBNA-1 antibodies titers were evaluated in blood of EBV-seropositive MS patients (n = 57; 31 relapsing remitting -RRMS- and 26 progressive -PMS-patients) and age- and sex-matched healthy controls (HC, n = 49). Results showed that EBNA-1 and VCA antibodies titers are significantly augmented in MS patients compared to HC (p < 0.05 for both antibodies); detection of EBV DNA was more frequent as well in MS patients compared to HC, although without reaching statistical significance. Regarding viral gene expression, LMP2A was significantly more frequently detected and more expressed in MS patients compared to HC (p < 0.005) whereas no differences were observed for EBNA-1. Considering patients alone, EBNA-1 was significantly more frequent in PMS compared to RRMS (p < 0.05), whereas no differences were observed for LMP2A. Increased expression of the LMP2A latency-associated gene in MS patients supports the hypothesis that EBV plays a role in disease etiopathology.

Investigating the role of Epstein-Barr virus and human papillomavirus types 16 and 18 co-infections in cervical disease of Iranian women.

High-risk human papillomaviruses (HR-HPVs) are known to contribute to cervical cancer (CC), but the role of Epstein-Barr virus (EBV) in this process remains unclear, despite EBV's widespread detection in premalignant and malignant cervical tissues. In this cross-sectional study of 258 cervical samples, including both formalin-fixed paraffin-embedded (FFPE) and fresh cervical tissues, the presence and viral load of HR-HPVs (HPV-16 and HPV-18) and EBV were evaluated in Iranian women with cervical intraepithelial neoplasia (CIN), squamous cell carcinoma (SCC), and a cervicitis control group using real-time PCR. The study revealed a significant correlation between disease severity and both increased HPV-16 positivity and HPV-16 and HPV-18 co-infection (p<0.001). Interestingly, the control group had a higher frequency of EBV-positive cases than SCC/CIN groups (p<0.001). HPV-16 DNA load increased with disease severity (P<0.001), while HPV-18 showed no significant difference (P=0.058). The control group had a higher EBV DNA load compared to SCC/CIN groups (P=0.033). HPV-16 increased the risk of CIN II, CIN III, and SCC, while HPV-18 increased the risk of CIN II and CIN III. Notably, EBV was associated with a lower risk of CIN groups and SCC. No significant difference in EBV co-infection with HPV-16/18 was found, failing to support the hypothesis that EBV is a cofactor in CC. However, high EBV viral load in the control group suggests a potential "hit and run hypothesis" role in CC progression. This hypothesis suggests that EBV may contribute briefly to the initiation of CC with an initial impact but then becomes less actively involved in its ongoing progression.