EBV PCR Research Articles

Global DNA Methylation Level in Tumour and Margin Samples in Relation to Human Papilloma Virus and Epstein-Barr Virus in Patients with Oropharyngeal and Oral Squamous Cell Carcinomas.

Aberrant DNA methylation is a common epigenetic modification in cancers, including oropharyngeal squamous cell carcinoma (OPSCC) and oral squamous cell carcinoma (OSCC). Therefore, the analysis of methylation levels appears necessary to improve cancer therapy and prognosis. The enzyme-linked immunosorbent assay (ELISA) was used to analyse global DNA methylation levels in OPSCC and OSCC tumours and the margin samples after DNA isolation. HPV detection was conducted by hybridisation using GenoFlow HPV Array Test Kits (DiagCor Bioscience Inc., Hong Kong, China). EBV detection was performed using real-time PCR with an EBV PCR Kit (EBV/ISEX/100, GeneProof, Brno, Czech Republic). OPSCC tumour samples obtained from women showed lower global DNA methylation levels than those from men (1.3% vs. 3.5%, p = 0.049). The margin samples from OPSCC patients with HPV and EBV coinfection showed global DNA methylation lower than those without coinfection (p = 0.042). G3 tumours from OSCC patients had significantly lower levels of global DNA methylation than G2 tumours (0.98% ± 0.74% vs. 3.77% ± 4.97%, p = 0.010). Additionally, tumours from HPV-positive OSCC patients had significantly lower global DNA methylation levels than those from HPV-negative patients (p = 0.013). In the margin samples, we observed a significant negative correlation between global DNA methylation and the N stage of OSCC patients (rS = -0.33, p = 0.039). HPV-positive OPSCC patients had higher global DNA methylation levels than HPV-positive OSCC patients (p = 0.015). We confirmed that methylation could be changed in relation to viral factors, such as HPV and EBV, as well as clinical and demographical parameters.

Targeting EBV Encoded Viral Kinases with GCV, AZT, Rituximab and Dexamethasone (GARD) Results in Durable Responses in Patients with CNS Lymphoproliferative Disease

Introduction: Epstein-Barr virus positive (EBV+) central nervous system lymphoproliferative diseases (CNS-LPD) are aggressive clinical conditions with poor prognosis. EBV+ CNS-LPD often occurs in the setting of immune suppression and comorbidities (solid organ transplant, autoimmunity). Treatment with high-dose methotrexate has increased survival but is not a viable option for all patients, highlighting the need for alternative treatments. Previous research has reported EBV+ CNS-LPD exhibits unique genetic and immunobiological signatures, however, few studies have reported on the epigenetic landscape. We have previously reported use of ganciclovir (GCV), zidovudine (AZT), rituximab and dexamethasone (GARD) regimen induced complete and durable responses in a cohort of 13 patients with primary CNS post-transplant lymphoproliferative disease (PCNS-PTLD). Here we present 24 patients (10 from prior cohort and 14 new) with EBV+ CNS-LPD treated with GARD. We extend follow-up time for the previous cohort and add molecular context to clinical observations. We performed DNA methylation and expression analysis of EBV viral kinases in available tumor samples and revealed unique viral epigenetic states leading to expression of antiviral drug targets in an array of EBV+ CNS-LPD. Methods: We conducted an IRB-approved, retrospective review of patients with EBV+ CNS-LPD treated with GARD at The Ohio State University Wexner Medical Center between 1998-2022. Patient information was extracted from the electronic medical record. Treatment consisted of two-week induction of twice daily IV GCV/AZT (5mg/kg and 1,500 mg) IV dexamethasone (10-40mg), and weekly rituximab (375mg/m 2). After induction, dexamethasone was tapered, and GCV/AZT was switched to maintenance oral dosing of 450mg valganciclovir twice daily and 300mg of AZT twice daily. Rituximab was administered on days 1, 8, 15, and 22. Two-year overall survival (OS), five-year OS, and overall response rate were assessed. Available CNS-LPD biopsy samples were obtained, and expression of EBV lytic genes BZLF1, BXLF1 (thymidine kinase), and BGLF4 (protein kinase) were measured via qRT-PCR. Mass spectrometry based EpiTYPER assay was used to report the percentage of DNA CpG methylation in the gene promoters for viral BZLF1, BXLF1, BGLF4 and LMP1 genes . Biopsies from 17 patients with systemic PTLD with no CNS involvement were used for comparison. Results: 24 patients with EBV+ CNS-LPD were identified. 21 (87.5%) patients were diagnosed with a primary CNS-LPD (PCNS-LPD). Of the PCNS-LPD, 14 patients were PCNS-PTLD, 4 were receiving immunosuppression for autoimmune conditions, 1 patient had type 2 diabetes and two had no known history of immunosuppression. The remaining three patients had systemic PTLD with secondary CNS involvement. Diffuse large B-cell lymphoma was the most common histology (11; 45.8%) followed by grade 3 lymphomatoid granulomatosis (5; 20.8%), B-cell LPD (5;20.8%), polymorphic PTLD (2;8.3%) and one large cell lymphoma of ambiguous lineage. All patients were HIV negative. EBER was positive in all biopsies and EBV PCR of cerebrospinal fluid was positive in patients without biopsy. Sixteen patients (66.7%) achieved a completed response, with an overall response rate of 83%. Two-year OS was 62.5% (95% CI, 45.8-85.2%) and five-year OS was 52.8% (95% CI, 35.7%-77.9%). Five CNS-LPD brain biopsies were available for analysis. qRT-PCR of biopsy specimens showed expression of LMP1, BXLF1, and BGLF4, but not BZLF1. Expression of BGLF4 was significantly higher (p=1.35e-5) in CNS-LPD when compared to systemic EBV+ PTLD. EpiTYPER data showed significantly decreased promoter methylation for viral kinases BXLF1 and BGLF4 (p=0.0281 p=0.0089). Conclusions: We have shown that GARD regimen shows efficacy in the treatment of EBV+ CNS-LPD, with 62.5% two-year OS and 52.8% five-year OS. Our results provide the molecular basis for expression of lytic viral kinases BXLF1 and BGLF4 in CNS-LPD by showing that both genes exhibit decreased promotor methylation. This expression of viral kinases in CNS-LPD supports the mechanistic rationale of this antiviral approach and could be further investigated as a predictive biomarker. Overall, this research highlights unique epigenetic features of EBV in CNS-LPD that support the use of antiviral, specifically the GARD regimen, in EBV+ CNS-LPD.

Epstein-Barr Virus Encephalitis: A Review of Case Reports from the Last 25 Years.

Although uncommon, Epstein-Barr virus-related neurological disorders represent the seventh most frequent cause of infectious encephalitis in adults. The limited number of publications on EBV encephalitis mainly document isolated clinical cases. This study aimed to summarize published data on EBV encephalitis. A systematic literature search identified 97 EBV encephalitis cases. In the selected cases, EBV-related neurological disorders manifested as lymphocytic pleocytosis in the cerebrospinal fluid (CSF) with moderate hyperproteinorachia. The EBV PCR test was positive in 87% of the CSF samples, with wide-ranging viral loads. When encephalitis occurred in the context of past EBV infections, all of the EBV PCR tests on CSF samples were positive. On the contrary, negative EBV PCR tests on CSF samples occurred only in the context of primary infections. EBV PCR was rarely carried out on blood samples, contributing minimally to the diagnosis. For the treatment of EBV encephalitis, Aciclovir was used alone in 29% of cases, and in association with other drugs in 40% of cases. Ganciclovir (30%), corticoids (52%), and immunoglobulins (15%) were mainly used in association with other drugs. Cerebral imaging was abnormal in 69% of cases, mostly in the cerebellum and basal ganglia. This work highlights that the EBV PCR test on CSF samples is currently the main laboratory diagnostic test to diagnose EBV encephalitis. This diagnostic test is useful; however, it is imperfect. New complementary diagnostic tools, approved treatments, and standardized practices could improve patient management.

Extranodal nasal-type NK/T lymphoma treated with chemotherapy and radiotherapy: case series from aEuropean tertiary referral center and review of the literature.

Extranodal nasal-type NK/T-cell lymphoma (ENKTL) is very rare in western countries and few data are available regarding the prognosis and the outcome of patients treated for this disease. We aimed to evaluate the prognosis, the pattern and risk factors of disease failure after combined therapy and also performed areview of the literature. We retrospectively analyzed 20patients with (ENKTL) who underwent L‑Asparaginase based chemotherapy followed by (chemo-) radiotherapy between 2010 and 2020 in our center. Data on clinical characteristics and irradiation were collected. Failure patterns were recorded as local (tumor site), regional (regional lymph nodes) or distant failure (metastasis and/or nonregional lymph nodes). During amedian follow-up period of 46months, disease failure was observed in 8patients (40%). The 3‑year progression-free survival (PFS) and overall survival (OS) rates were 62.5 and 83.0%, respectively. The failure patterns were local (n = 6, 30%), regional (n = 3, 15%) and distant (n = 4, 20%). Among patients with local failure, all failures occurred within the radiation fields (100%). Univariate analysis showed that bilateral regional lymph node involvement (p = 0.0002), initial circulating EBV viral load ≥ 3.5 log (p = 0.03) and no negativation of EBV PCR after induction CT (p = 0.0497) were independent predictors of PFS. Patients with bilateral lymph node involvement and/or high EBV viral load have asignificant recurrence rate despite multimodal therapy. These results need to be confirmed by larger studies. Given the high rate of local recurrence within radiotherapy fields, the value of dose escalation should be considered. Patients at risk of relapse should be included in dedicated trials.

The Impact of Epstein-Barr Virus Infection on Second Cancer Development in Newly Diagnosed Myeloma Patients Treated with Lenalidomide in the UK NCRI Myeloma XI Trial

Background Several factors have been linked to second primary malignancy (SPM) development in myeloma patients, including advanced age, immunocompromise and therapy, such as melphalan and lenalidomide (len). The impact of infection, including EBV is yet to be elucidated. Here we address this in the context of the UK NCRI Myeloma XI Trial for newly diagnosed transplant eligible (TE) and transplant non-eligible (TNE) patients. Methods Myeloma XI was a phase III, randomised, multi-centre, parallel group design, open-label trial comparing cyclophosphamide, thalidomide, len, carfilzomib and bortezomib induction combinations and len maintenance treatment in TE and TNE patients. TE patients received high dose melphalan supported by autologous stem cell transplantation (ASCT). Patients in both pathways were randomised to maintenance with len or observation. In total, 4358 patients entered. Planned SPM analysis was conducted in May 2019, with 318 patients developing an SPM prior to that point. Median follow-up from trial entry was 60 months (IQ range 47-76). A nested cohort analysis was performed to determine EBV infection and immunity status in 170 SPM patients for which serum samples were available. Forty-two trial patients who did not develop an SPM were used as a control group. EBV assessment time-points included pre-induction, post induction, 4 months post ASCT (TE only) and 1 year post induction. Peripheral blood serum samples were tested for EBV EBNA IgG Antibody/index (LIAISON®), EBV VCA IgM Antibody/Index (LIAISON®) and EBV positivity was determined using the artus® EBV RG PCR Kit from Qiagen. Active or recent EBV infection was evidenced by a positive EBV IgM antibody and/or EBV PCR. Results Of the 170 patients, 208 SPM were noted with 147 patients developing 1 SPM and 23 patients developing ≥2. One hundred and five patients developed an SPM following maintenance randomisation, with 76 receiving len and 29 being observed. The remaining 65 patients developed an SPM prior to maintenance. Eighty-four SPMs were diagnosed in 68 TE patients and 124 SPMs in 102 TNE patients. Of the 170 SPM patients, 8 had evidence of recent or active infection at the time of testing, evidenced by either IgM EBV antibodies (n=5), positive EBV PCR (n=2) or both (n=1). The median EBV DNA copies/ml was 353 (range 337-428). The median IgM antibody titre was 71.5U/ml (26 - >160). No patients in the control group had evidence of recent EBV infection (p=0.36). Of the 8 patients with evidence of EBV infection, 6 developed >1 SPM, Table 1. Five had been treated in the TE pathway, with all developing post maintenance randomisation, and 4 of the 5 receiving len maintenance. None of the 3 TNE patients received len maintenance, with 2 developing an SPM prior to maintenance randomisation. The median time from EBV testing to SPM diagnosis was 17·9 months (0.5-51.7). Two of the EBV positive patients developed a haematological SPM: myelodysplastic syndrome and Hodgkin lymphoma. Four patients developed a solid malignancy: lung, breast, ovary, and colon. The remaining cases were non-melanoma skin cancers, Table 1. Immune status (EBV IgG) was also determined. In TE patients, the median EBV IgG titre was 65.3U/ml at diagnosis (<3 - 600, n=48), falling to 28.1U/ml following induction (<3 - 326, n=41) and recovering to 44.2U/ml one year post ASCT (<3 - 600, n=22). In TNE patients, the median IgG titre at diagnosis was 39.1U/ml (<3 to 517, n=72), falling to 31.2U/ml 4 months post induction (<3 - 287, n=42) and recovering to 51.3U/ml after 1 year (<3 - 554, n=41). In the SPM patients the IgG EBV titres were noted to be <20U/ml in 5 of the 8 patients. Three of the eight patients had no detectable EBV IgG level (<3U/ml) Conclusions These data suggest that EBV infection is unlikely to be a risk factor for SPM development in myeloma patients. This is supported by the finding that EBV IgG titres were consistent with immunity in most patients (>20U/ml) and even in the patients with possible EBV infection, EBV DNA was only identified in 3 patients, and at a low level. In addition, the SPM developed are not classically associated with EBV infection or reactivation. It is noted that EBV infection status was not always determined at the time of SPM development. Despite this, the data does show that EBV immunity recovers following therapy, even in the post ASCT setting. EBV monitoring is therefore not supported by this work, although in patients who develop ≥2 SPM, further clarification is required.

Evaluation of two automated real-time PCR-based quantification methods for whole blood Epstein-Barr viral load

Quantification of EBV DNA is important in transplantation settings for the diagnosis of post-transplantation.We evaluated the performance of the AltoStar® EBV PCR Kit 1.5 on whole blood specimens: limit of detection, linearity, accuracy, and precision were determined using the WHO NIBSC 09/260 international standard. Results of 69 clinical samples were compared between the AltoStar® EBV PCR Kit 1.5 (altona Diagnostics) and the RealTime EBV assay (Abbott).The LoD of the AltoStar® Kit was 148 IU/mL and linearity was between 375 and 500000. A high concordance was found between nominal value of the NIBSC dilutions and the AltoStar EBV result. The total variation ranged from 2.2% to 9.6%.Out of 69 clinical samples tested, there was a high concordance between the 22 paired results within the overlapping linear ranges of both tests.The AltoStar® EBV assay is reliable and accurate for EBV viral load determination on whole blood samples.

An Atypical Presentation of X-linked Lymphoproliferative Disease type 1 Presenting with Seronegative Coccidioidomycosis

X-linked lymphoproliferative disease (XLP-1), an inborn error of immunity (IEI) caused by pathogenic variants in SH2D1A encoding signaling lymphocyte activation molecule-associated protein (SAP), is characterized by hemophagocytic lymphohistiocytosis (HLH), lymphoma, and hypogammaglobulinemia. We report the first known case of XLP-1 presenting with seronegative coccidioidomycosis. CaseA 13-year-old boy with persistent asthma, recurrent otitis media requiring four sets of tympanostomy tubes, chronic sinusitis, and Common Variable Immunodeficiency (CVID) presented with acute-on-chronic hypoxic respiratory failure. He had been off subcutaneous immunoglobulin (SCIg) replacement for 2 years. Sputum culture was positive for H. influenzae, treated with cefepime and azithromycin and supplemental oxygen for acute hypoxic respiratory failure. Fever persisted despite 48 hours of antibiotics. CT chest showed bronchiectasis with mediastinal, subcarinal and hilar adenopathy. Coccidioides immitis IgG and IgM were negative, but urine antigen was positive. There was no radiologic or cerebrospinal fluid evidence of disseminated coccidioidomycosis.Immunologic workup revealed undetectable IgG, IgA, IgM, and IgE (table 1). Lymphocyte subsets showed severe T and NK cell lymphopenia. Total B cell count was normal, but he had decreased CD19+27+ and transitional B cells, with absent switched memory B cells and no plasmablasts (table 1). Lymphocyte proliferation to mitogens/antigens was decreased. Serum ferritin was normal, and EBV PCR was negative. A targeted 429-gene panel identified a hemizygous variant of uncertain significance (VUS) in SH2D1A, c.82_102dup (p.Ser28_Ser34dup). SAP expression was absent in CD4+ T cells and NK cells with a bimodal expression in CD8+ T cells, suggesting possible mosaicism in this subset. He was diagnosed with X-linked lymphoproliferative disease 1 (XLP-1) and treated with fluconazole, trimethoprim-sulfamethoxazole prophylaxis, and IgG replacement (1 g/kg intravenous initially, then transitioned to weekly SCIg targeting an IgG >1000 mg/dL), with clinical improvement, resolution of fevers and acute hypoxia. He was referred for hematopoietic cell transplant evaluation.Coccidioidomycosis has not been described in a new diagnosis of XLP-1 and should be considered in this IEI. Coccidioidomycosis susceptibility has been reported in other IEIs including impaired interleukin-12/interferon-gamma axis signaling, STAT3 loss of function, chronic granulomatous disease, and recently cytidine triphosphate synthase 1 (CTPS1) deficiency.Table 1Patient’s immunologic variables compared to reference range. Absolute values are shownImmunologic VariablePatient ValuesReference RangeIgG<70 (L)<70 mg/dLIgA<15 (L)<15 mg/dLIgM<8 (L)<8 mg/dLIgE<1.5 (L)<1.5 IU/uLTotal CD4+ T Cells11 (L)570–2400 cells/uLTotal CD8+ T Cells63 (L)210–1200 cells/uLTotal CD19+ B cells173165–1154 cells/uLCD19+ CD24++ CD38++ (Transitional B Cells)0 (L)8–392 cells/uLCD19+CD24+CD38+ (Naïve B Cells)12159–1146 cells/uLCD19+CD27-(Naïve B cells)169140–2269 cells/uLCD19+ CD27+ CD38+ IgM- (Plasmablasts)0<30 cells/uLCD19+CD27+IgD-IgM- (Switched-memory B cells)0 (L)6–190 cells/uLCD16+ CD56+ (NK Cells)32 (L)78–470 cells/uLL – low

EBV-related lymphoproliferative disease in Wiskott-Aldrich Syndrome patient

CaseWiskott-Aldrich Syndrome (WAS) is a rare X-linked inborn error of immunity caused by mutations in the WAS gene. It is classically characterized by immunodeficiency, eczema, and micro-thrombocytopenia. It has been known since the 1960s that patients with WAS have an increased risk of lymphoproliferative disease though the exact incidence remains unknown in the American population. Limited case reports have discussed EBV-related lymphoproliferative disease in patients with WAS. We present a case of a 9-year-old boy with known WAS complicated by eczematous rash, thrombocytopenia, recurrent ear infections, and monoclonal gammopathy who was found to have submandibular EBV-associated lymphoid hyperplasia with associated lung and retroperitoneal lymphadenopathy. Family had been offered treatment with hematopoietic stem cell transplant but declined multiple times in the past. Earlier in the year, he presented with possible MIS-C with negative SARS-CoV-2 PCR. He presented to our hospital with mastoiditis and lymphadenopathy. Physical examination showed severe eczema on hands and tender right mastoid. Laboratory evaluation showed thrombocytopenia, elevated IgG of 6290, IgA of 744, IgE of 827, low IgM of 41, and 14% response to pneumococcal titers. He was empirically treated with intravenous antibiotics. ENT performed right postauricular incision and drainage and the culture grew Hemophilus influenza. Throughout his hospital stay, his submandibular lymphadenopathy became more prominent despite treatment. Core needle biopsy of right submandibular lymph node was suggestive of EBV-associated lymphoid hyperplasia. EBV PCR and antibodies were both positive. CT chest, abdomen, and pelvis revealed multifocal pulmonary lymphadenopathy and a diffuse, bilateral nodularity as well as retroperitoneal and mesenteric lymphadenopathy. He was given four doses of weekly Rituximab, which successfully decreased EBV viremia below linear detectability. Immunoglobulin replacement therapy (IgRT) was initiated. Bronchoalveolar lavage and lung biopsy were performed and are results are currently pending. DiscussionWe present a case of a 9-year-old boy with known WAS awaiting transplant who was found to have submandibular EBV-associated lymphoid hyperplasia with associated lung and retroperitoneal lymphadenopathy. While lymphoproliferative disease is a known complication of WAS, EBV-related lymphoproliferative disease in WAS patients has only been reported as case reports and remains a rare but known complication of patient with WAS.

T-cell chronic active EBV diagnosed in a 65-year-old male presenting as idiopathic hypereosinophilic syndrome

A 65-year-old male from Nigeria, admitted with dehydration and painful oral ulcers, is seen by allergy/immunology for hypereosinophilia.The patient had idiopathic eosinophilia (mean 5,000 cells/uL) ongoing for nine years. There were no evident drug or infectious causes. The eosinophilia peaked at 28,000 cells/µL, accompanied by parotid swelling, pustular skin rash, and elevated serum-IgG4 level (400 mg/dL). Rheumatology evaluated for IgG4-related disease and hypereosinophilic syndrome. This work-up demonstrated elevated IgE (39,000 IU/mL) and tryptase (40 ng/mL) levels. Bone marrow biopsy showed a hypercellular marrow with erythroid hyperplasia and eosinophilia. Submandibular gland, skin, and excisional lymph node biopsies demonstrated IgGsecreting plasma cells, lymphocytic/eosinophilic infiltrates, and lymphoid hyperplasia, respectively. All biopsies stained negative for IgG4. Treatment for idiopathic hypereosinophilia was initiated with corticosteroids, rituximab, and mycophenolate mofetil (MMF). Symptomatic improvement was attained on MMF (1000 mg twice daily) with the addition of mepolizumab (300 mg every 4 weeks).He unfortunately progressed to high fevers with elevated ferritin (7,286 ug/L) and soluble IL-2Ra (31,276.7 pg/mL) levels. He did not meet full criteria for hemophagocytic lymphohistiocytosis and improved after corticosteroids. EBV PCR from blood was >220,000 IU/mL.Dedicated immune work-up showed a non-specific population of lymphocyte-like cells without clear lineage markers (CD2dim/- surface CD3-CD4-CD8-CD7-CD56-CD49d-CD294dim) on peripheral blood flow cytometry, with otherwise normal T, B, and NK cell enumeration. There was reduced cytotoxic killing function of CD8+ T cells and NK cells by chromium-51 release assay. Sorted peripheral blood mononuclear cells demonstrated positive EBV viral load in B cells (1,018,239 IU/mL), T cells (857,418 IU/mL),NK cells (237,459), and myeloid cells (532,641 IU/mL). EBV-encoded RNA was positive within lymph node T cells, most consistent with a diagnosis of T-cell CA-EBV. The Invitae 407 gene immunodeficiency panel was sent; however, a pathogenic variant was not identified. The patient was seen by hematology/oncology with plans for hematopoietic stem cell transplant. Unfortunately, he succumbed to complications from CA-EBV and multiorgan failure.We describe a male with lymphocytic-variant hypereosinophilic syndrome and a longstanding constellation of inflammatory and lymphoproliferative symptoms due to CA-EBV. His course initially eluded his care team and only after careful multidisciplinary discussion was a diagnosis reached.

S3106 N-Acetyl Cysteine Use in the Treatment of Ischemic Hepatitis in the Setting of COVID-19 Infection

Introduction: Acute liver injury occurs in 15-53% of COVID-19 patients. Most often, elevation in transaminases is mild, but severe liver injury has been described possibly secondary to hepatic ischemia. We present a case of ischemic hepatitis with concurrent COVID-19 infection, in which N-acetyl cysteine (NAC) administration led to clinical improvement and recovery. Case Description/Methods: A 58-year-old male with adult congenital heart disease (ACHD), on warfarin for atrial flutter and cardiac cirrhosis presented to the ER with generalized fatigue, chills, congestion, and cough. Patient tested positive for COVID-19. He was hemodynamically stable with an unremarkable exam. He was alert and oriented times three. He initially required 8 L of oxygen which continued to improve on a daily basis. His initial laboratory workup was notable for AST 1954 U/L, ALT 1905 U/L, INR 19.1, and total bilirubin 0.9 mg/dL. At baseline, patient’s liver enzymes were normal. He was admitted to the ICU due to the severity of his liver injury. Alcohol, acetaminophen, salicylate levels and urine drug screen were all within normal limits. Abdominal ultrasound revealed known liver cirrhosis and ascites. Acute hepatitis serologies including hepatitis E IgM, ANA, AMA, ASMA, anti-LKM, IGG subclasses, ceruloplasmin, alpha-1 antitrypsin, and celiac panel were done and unremarkable. In addition, HIV, VZV PCR, EBV PCR, HSV PCR and CMV PCR were also negative for acute infection. Patient was treated with a NAC protocol with decrease in elevated liver enzymes by half in the first 72 hours and >20-fold of the highest level upon discharge. Throughout his admission, his mental status remained normal. Discussion: Ischemic hepatitis (IH) results from insufficient blood flow volume and/or oxygen content to the hepatocytes. Acute hypoxic respiratory failure as a result of COVID-19 infection is one of the potential causes of this type of liver injury. Other possible causes of liver injury include direct viral cytopathic effect, cytokinesis and drug-induced liver injury. In the setting of COVID 19 infection, prompt diagnosis and recognition of IH is critical as studies describe mortality rates as high as 50%. We describe our clinical experience with NAC in the setting of COVID 19 with severe IH and propose that NAC is considered in the treatment of COVID-19 patients with ischemic hepatitis, however further research including prospective clinical trials is needed to better validate this.

ACUTE FIBRINOUS AND ORGANIZING PNEUMONIA (AFOP) PRESENTING AS DIFFUSE CAVITARY LESIONS IN THE SETTING OF EPSTEIN BARR VIRUS (EBV)-INDUCED HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS (HLH)

ACUTE FIBRINOUS AND ORGANIZING PNEUMONIA (AFOP) PRESENTING AS DIFFUSE CAVITARY LESIONS IN THE SETTING OF EPSTEIN BARR VIRUS (EBV)-INDUCED HEMOPHAGOCYTIC LYMPHOHISTIOCYTOSIS (HLH)

S3064 Intestinal Post-Transplant Lymphoproliferative Disorder

Introduction: Post-transplant lymphoproliferative disease (PTLD) is a group of lymphoid disorders arising in the setting of immunosuppression following a solid organ or stem cell transplantation. PTLD impacts 2.8% of adults and 15% of children post-orthotopic liver transplant (OLT) at a median of 8 years. The small intestine, rich in B cells, is a common site for PTLD and the treatment is chemotherapy. Here, we present a case of a patient post-OLT presenting with abdominal pain, and anemia found to have small bowel PTLD resulting in jejunal perforation with eventual resolution of symptoms after surgical resection and chemotherapy. Case Description/Methods: 65-year old obese F with non-alcoholic steatohepatitis decompensated by hepatopulmonary syndrome with O2 dependence and hepatocellular carcinoma underwent an uneventful OLT (CMV +/-) with no immediate postoperative complications and resolution of the hepatopulmonary syndrome. Presented 2 months after the OLT with worsening LLQ abdominal pain and acute anemia. An EGD and colonoscopy completed shortly after admission showed evidence of gastric, duodenal, and descending colon non-bleeding ulcers that were biopsied. Pathology showed EBV+ atypical B cell proliferation concerning for large B cell lymphoma and monomorphic-PTLD. An EBV viral load obtained at that time was elevated and subsequent CT Abdomen/Pelvis showed scattered small sub-centimeter mesenteric and retroperitoneal lymph nodes. Her immunosuppression was minimized without a change in symptoms. She subsequently developed large volume haematochezia leading to shock and underwent an EGD and colonoscopy followed by a push enteroscopy that revealed multiple ulcers but no active bleeding site. Immediately after the procedure, she developed an acute abdomen due to a perforated Jejunal ulcer. Had an emergent exploratory laparotomy and jejunectomy with subsequent improvement in her anemia. She was treated with R-CHOP for PTLD with the resolution of her presenting symptoms. Discussion: PTLD, however rare, remains the commonest post-transplant malignancy. Once there is suspicion (pain, weight loss, and anemia post OLT), EGD/Colonoscopy reveals suspicious malignant-looking ulcers, and mucosa or lymph node biopsy confirms PTLD. The patient ultimately underwent treatment with 4 cycles of R-CHOP with surveillance EBV PCR and imaging with resolution of complete symptoms. Immediate treatment should be started with lowering immune suppression and chemotherapy rather than waiting for self-resolution in PTLD cases.Figure 1.: Jejunal and Gastric ulcers (seen on prior scopes as well) were found on the Push enteroscopy just prior to the perforation of the bleeding jejunal ulcer and eventual surgical resection of the jejunum that resolved her anemia.

The relationship between tumour infiltrating lymphocytes, Epstein-Barr virus and Helicobacter pylori infection in gastric cancer.

ObjectiveEpstein–Barr virus (EBV) and Helicobacter pylori (HP) infections have been extensively recognised as gastric cancer (GC) triggers, and recent publications suggest they could behave as predictive markers for immune-modulating therapies. Tumour-infiltrating lymphocytes (TILs) have also been identified as a predictive biomarker for immunotherapy in different malignancies. This study aimed to investigate the association between EBV and HP infection with TIL levels in GC.MethodsTIL evaluation in haematoxylin-eosin was performed by a pathologist and density of CD3, CD8 and CD163 positive (immunohistochemistry staining) immune cells was calculated with the use of digital pathology software. EBV infection was detected by in situ hybridisation (ISH) and by quantitative polymerase chain reaction (qPCR). Methylation status of EBV-related genes was detected by PCR and a methylome analysis was performed by the Illumina Infinium MethylationEPIC BeadChip. HP status was detected by qPCR.ResultsWe included 98 resected GC Peruvian cases in our evaluation. Median TIL percentage was 30. The proportion of EBV+ detected by ISH was 24.1%, of EBV+ detected by qPCR was 41.8%, while 70% showed methylation of EBV-related genes, and 58.21% of cases were HP+. Younger age (p = 0.024), early stages (p = 0.001), HP+ (p = 0.036) and low CD8 density (p = 0.046) were associated with longer overall survival (OS). High TIL level was associated with intestinal subtype (p < 0.001), with grade 2 (p < 0.001), with EBV qPCR+ (p = 0.001), and with methylation of EBV-related genes (p = 0.007). Cases with high TIL level and cases that are EBV positive share eight genes with similarly methylated status in the metabolomic analysis. High CD8 density was associated with EBV PCR+ (p = 0.012) and HP− (0.005).ConclusionLower CD8 density and HP+ predict longer OS. High TIL level is associated with EBV+ and methylation of EBV-related genes, while lower CD8 density is associated with HP+ GC.

EBV Positive DLBCL Is Effectively Treated with Commercial CAR T Cell Therapy but Is Associated with Higher Rates of Severe Immune Effector Cell Associated Neurologic Syndrome (ICANS): A Single Institution Experience

Introduction:CAR T-cell therapy (CART) has revolutionized the treatment landscape of aggressive B cell lymphoma (aBCL) in the relapsed/refractory (r/r) setting, including high-risk groups such as high grade B cell lymphoma (HGBCL), primary refractory disease, and chemo-resistant relapses. To date, however, there has been no data reported, on the outcomes of patients with Epstein-Barr virus positive (EBV+) DLBCL, NOS. EBV+ DLBCL, NOS is a high-risk group of aBCL characterized by the incorporation of EBV into the malignant cells, with the chronic inflammation associated with EBV infection being thought to contribute to DLBCL pathogenesis. It has been shown to be less responsive to standard chemotherapy with high relapse rates and overall poor prognosis. In the present study, we analyze outcomes and toxicities of patients with EBV+ DLBCL, NOS treated with CD19CART.Methods:We retrospectively analyzed charts of patients treated with commercially available CD19CAR T-cell products for the treatment of aBCL at City of Hope (COH) between April 2018 and March 2021. EBV status was determined by immunohistochemical stains from EBER in situ hybridization on paraffin embedded tissues. Patients were included in the analysis if they had positive or negative EBER staining, referred as EBV+ or EBV-, respectively. The investigation was approved by the COH IRB. Cytokine release syndrome (CRS) and immune effector cell associated neurologic syndrome (ICANS) were graded using ASTCT criteria (Lee et al. BBMT 2019). Demographics were analyzed with descriptive statistics. Univariate analysis was performed by Chi Square test.Results:Seventy-three patients had tumor samples that were evaluable for analysis, 92% (n=67) were negative for EBER and 8% (n=6) were positive for EBER. In the entire cohort, the median age was 62 years (range, 21-90), and included 18% (n=13) transformed follicular lymphoma, 73% (n=53) DLBCL, NOS, and 10%(n=7) DH. The median number of prior lines of therapy was 2 (range, 2-6) and 15% (n=11) patients had prior autologous stem cell transplant.Comparing the EBV+ patients to the EBV-, the CR rate was 67% (n=4) vs 58% (n=34), with 16% (n=1) PR vs 27% (n=15) (ORR 83% vs 74%). With a median follow up of 17.0 months (range, 3.6-38.3), 33% (n=2) of EBV+ patients had progressed vs 42% (n=28). In the EBV+ DLBCL, NOS group, 3 CR patients had absence of detectable EBV PCR in the peripheral blood at the time of CART; 1 CR patient did not have clinical indication for EBV viral load (VL) evaluation (EBV PCR not measured); 1 PR and 1 PD patients had detectable viremia. At the time of analysis, 31% (n=21) of EBV- patients died vs 33% (n=2) of EBV+ patients, both with non-CR response to CAR T and persistent EBV VL during therapy. In terms of toxicity, overall CRS rates were similar with 83% (n=5) in the EBV+ patients and 86% (n=58) in EBV- patients. Although overall incidences of ICANS were similar between the two groups (50% [n=3] vs 45% [n=30]), grade 3 ICANS was higher in EBV+ patients (50% [n=3]) than in EBV- patients( 7.5% [n=5]; p = 0.02). EBV VL did not correlate with CRS or neurotoxicity.Conclusion:We present a retrospective analysis of EBV+ DLBCL, NOS patients treated with CART at our institution. Based on our analysis, despite the limited number of cases, the CART is an effective therapy for EBV+ DLBCL,NOS with response rates similar to what has been reported in the literature for other aBCL groups. Responses appear to be improved if EBV VL is undetectable at the time of CART. Interestingly, we demonstrate a statistically significant higher rates of grade 3 ICANS in patients with EBV+ DLBCL,NOS. Our data suggests that close monitoring for ICANS during CART should be considered in EBV+ DLBCL patients with early therapeutic intervention to prevent severe toxicity. Further investigation of a larger cohort of EBV+ DLBCL patients, as well as a deeper analysis of inflammatory markers and EBV viremia in these patients undergoing CART may provide further insight to response and toxicity profile. DisclosuresNikolaenko: Rafael Pharmaceuticals: Research Funding; Pfizer: Research Funding. Herrera: Seagen: Consultancy, Research Funding; AstraZeneca: Consultancy, Research Funding; Genentech: Consultancy, Research Funding; Takeda: Consultancy; ADC Therapeutics: Consultancy, Research Funding; Merck: Consultancy, Research Funding; Bristol Myers Squibb: Consultancy, Research Funding; Tubulis: Consultancy; Kite, a Gilead Company: Research Funding; Karyopharm: Consultancy; Gilead Sciences: Research Funding. Budde: Mustang Bio, Inc: Research Funding; Gilead: Consultancy; Merck, Inc: Research Funding; AstraZeneca: Research Funding; Amgen: Research Funding; IGM Biosciences: Research Funding; Roche: Consultancy; BeiGene: Consultancy; Novartis: Consultancy. Shouse: Kite Pharma: Speakers Bureau; Beigene: Honoraria.

S2679 An Unusual Suspect for Cholestatic Liver Injury

Introduction: Vanishing Bile Duct Syndrome (VBDS) and idiopathic cholestasis (IC) have been associated with lymphoma in rare cases. In VBDS, the defining feature on liver biopsy is ductopenia, while in IC there is canalicular cholestasis and mixed inflammatory changes. Existing literature reports a few cases of paraneoplastic cholestasis with pathological features of either VBDS or IC. We describe an unique case of severe cholestasis as the presenting sign of Hodgkin’s lymphoma in the absence of bile duct loss or inflammatory changes. Case Description/Methods: 60-year-old man with no past medical history presented to the ED with increasing fatigue and jaundice that started five months earlier. He underwent cholecystectomy and intraoperative liver biopsy four months earlier which showed bland lobular cholestasis and mild lobular injury with no fibrosis, inflammatory changes, abnormal lymphocytic infiltration, or evidence of bile duct loss. An MRI/MRCP 6 weeks before presentation showed multiple small hepatic cysts and no biliary obstruction or ductal dilatation. Ursodiol 14 mg/kg/d was started 2 weeks later but he developed hypotension and presented to the ED. In the ED, vital signs were normal. Physical exam showed jaundice and hepatomegaly. Labs revealed ALP 1002 U/L, AST 81 U/L, ALT 90 U/L, total bilirubin 10.8 mg/dL, and direct bilirubin 8.1 mg/dL. Further work up showed negative AMA, ANA, ASMA and vasculitis serologies, normal ceruloplasmin, and negative viral studies including hepatitis A, B, C, HSV, CMV, HIV and COVID-19. EBV PCR was 69,697 copies/mL with positive IgG suggestive of reactivation of previous infection. Lyme and Ehrlichia serologies were negative. Repeat MRI/MRCP showed normal bile ducts but extensive retroperitoneal lymphadenopathy. Lymph node core needle biopsy revealed classic Hodgkin’s lymphoma. He was started on chemotherapy and liver enzymes improved. Discussion: Hodgkin’s lymphoma can present with cholestatic liver injury as shown in our patient who was on no medications and had no other identifiable causes of cholestasis. In the existing literature, VBDS and IC have been described as the pathological features for these patients where either ductopenia or canalicular cholestasis and mixed inflammatory changes are seen. Our case is unique because our patient had neither of these features. Bland cholestasis associated with lymphoma should be kept in the differential in someone with otherwise unexplained cholestatic liver injury.Figure 1.: Panel A: High power view of liver biopsy (200x, H&E). Yellow-green bile can be seen in dilated canaliculi. There is no significant lobular inflammation, steatosis, or other alterations. Panel B: High power view of liver biopsy (400x, H&E). Pale yellow-green watery bile can be seen in dilated canaliculi (yellow arrows). There is no significant lobular inflammation, steatosis, or other alterations. Panel C: High power view of lymph node (200x, H&E). The lymph node shows loss of normal architecture with replacement by fibrosis and an atypical lymphohistiocytic infiltrate. Scattered large cells with irregular nuclear contours and hyperchromasia are seen suggestive of Hodgkin cells. Panel D: Immunoperoxidase studies show the Hodgkin Reed-Sternberg (HRS) cells are positive for CD15 and CD30. PAX-5 is weakly expressed in the large cells in comparison to the background small B cells. CD20 stains background small B cells but is negative in the HRS cells. The immunophenotype is consistent with classic Hodgkin lymphoma.

Use of Rituximab for persistent EBV DNAemia, and Its effect on donor-specific antibody development in pediatric renal transplant recipients: A case series.

Persistent EBV DNAemia (PEBV) is associated with late-onset PTLD. The efficacy of rituximab in PEBV is not conclusive. We monitored PEBV and DSA in pediatric kidney transplant patients with or without rituximab. 13 PEBV patients received standard treatment with immunosuppression reduction and valganciclovir, with or without IVIG; 5/13 were further treated with rituximab. All Rituximab-treated and 6/7 No-Rituximab patients were EBV seronegative at transplant and seroconverted post-transplant. Peak EBV PCR levels were lower in No-Rituximab than Rituximab patients and all No-Rituximab patients cleared PEBV after standard treatment. Additional 1-2 doses of rituximab reduced EBV PCR levels in all 5 Rituximab patients, 3 cleared PEBV. One No-Rituximab patient developed localized PLTD. None of Rituximab patients developed de novo DSA, while 4/8 No-Rituximab patients did: 2/4 had ABMR. 1/5 Rituximab and 5/8 No-Rituximab patients had acute rejection. There was no change in eGFR between pre-EBV DNAemia and follow-up in Rituximab patients, while reduction in No-Rituximab patients was found. There was no difference in graft and patient survival. While early intervention with rituximab in pediatric patients with PEBV may reduce viral load and PTLD, we observed a slower development of de novo DSA, and rejection and maintenance of eGFR.

POS-755 POST-TRANSPLANT LYMPHOPROLIFERATIVE DISORDER (PTLD) IN A 26 YEAR OLD FEMALE RENAL RECIPIENT - WHAT YOU CAN LEARN FROM OUR EXPERIENCE

PTLD accounts for almost 20% of all cancers occurring in solid organ transplantation. It represents infiltration of B cells driven by EBV/CMV infection in the background of severe immunosuppression and impaired T-cell immune surveillance. Current case presents PTLD 7 months post 2nd renal transplantation and reflects on early detection/prevention and management. Case Presentation A 26-year old female presented with 2/52 fatigue, fever, night sweats and abdominal pain, 7 months post 2nd cadaveric renal transplant EBV/CMV mismatched. Medical history included cyclophosphamide-treated proliferative lupus nephritis, tacrolimus-based immunotherapy and antirejection treatment. On examination, there was tenderness of the right iliac fossa with femoral lymph nodes. Patient had anaemia, leukopenia, deterioration in renal function, elevated LDH, EBV PCR 18,752 copies/mL and CMV PCR 300 copies/ml. CT abdomen showed an 8cm soft tissue mass obstructing the graft. Biopsy confirmed polymorphic PTLD (pictures 1, 2). View Large Image Figure ViewerDownload Hi-res image Download (PPT)View Large Image Figure ViewerDownload Hi-res image Download (PPT) Outcome: The patient was treated with reduction of immunosuppression and withdrawal of both Tacrolimus and Mycophenolate Mofetil. Prednisolone 5 mg daily remained and EBV load monitored weekly. Immunotherapy with CD 20 monoclonal antibody, rituximab, commenced weekly for 4 weeks. Complete remission was achieved in 3 months, Kidney function returned to baseline and 18F Fluorodeoxyglucose PET-CT revealed no sign of lymphoma. Four-year post diagnosis, patient remained disease-free as demonstrated on repeat 18 FDG PET- CT, with stable graft function and negative EBV/CMV viral load. Immunosuppression remained minimal. Seronegative EBV/CMV recipients accepting seropositive grafts have 24 times higher incidence rate of PTLD, likely within the 1st year post transplantation. Significant increase in the viral load can predict/diagnose early PTLD. Lower immunosuppression with reduction/withdrawal of Tacrolimus and rituximab use can be effective for prevention and treatment. Ganciclovir has proved to be of equal value. FDG PET-CT is highly sensitive to detect disease extent and response to treatment.

P0494UNUSUAL CASE OF ACUTE KIDNEY INJURY (HISTOLOGICAL EVALUATION: COLLAPSING GLOMERULOPATHY) IN HEMOPHAGOCYTIC SYNDROME ASSOCIATED WITH NK/T LYMPHOMA

Abstract Background and Aims Hemophagocytic lymphohistiocytosis (HLH) is a severe hyperinflammatory syndrome induced by aberrantly activated macrophages and cytotoxic T cells. The primary (genetic) form, caused by mutations affecting lymphocyte cytotoxicity and immune regulation, is most common in children, whereas the secondary (acquired) form is most frequent in adults. Secondary HLH is commonly triggered by infections or malignancies but may also be induced by autoinflammatory/autoimmune disorders, in which case it is called macrophage activation syndrome. The diagnosis of HLH in adults should be based on the HLH-2004 diagnostic criteria in conjunction with clinical judgment and the patient’s history. Renal involvement has previously been reported in 24 adult cases, mostly as acute renal failure. Collapsing glomerulopathy is extremely rare with only six previous cases reported in the literature. Case presentation We report the case of an African man, 31 years old, presented with fever, acute kidney injury: serum creatinine 10.3 mg/dl; urine protein 600 mg/dl, macrohematuria, ANA/ANCA were negative, low serum C3, organomegaly, anemia, thrombocytopenia, hypertriglyceridemia, hypofibrinogenemia, hyperferritinemia, direct and indirect antiglobulin (Coombs) tests were negative, low haptoglobin; elevated LDH; normal partial thromboplatin time. Peripheral blood smear examination reveal few schistocytes. ADAMTS13 activity was found to be 25%. HBV-DNA and HIV were negative. Anticardiolipin antibodies were negative. Lab exam suggested the relapse of an EBV infection and primary mycoplasma infection. Because of uremic symptoms and persisting oliguria we started replacement therapy by hemodialysis. Plasmapheresis was started because of suspected thrombotic microangiopathy. Suprisingly the kidney biopsy was consistent with collapsing glomerulopathy with evidence of tubular injury while the bone marrow biopsy diagnosed an EBV NK/T-Cell lymphoma. During the course of his hospitalization, the patient suffered high fever. C-reactive protein, WBC and procalcitonin levels were elevated. Antimicrobial agents were initiated, starting with ceftriaxone then upgraded to piperacillin/tazobactam and then the shifted to teicoplanin and meropenem. Blood, urine and stool cultures were negative.VRE positive, IgM Mycoplasma pneumoniae were positive; EBV PCR on bone marrow blood was positive. Malaria screening was negative. The antibiotic therapy was finally switched to doxycycline as unique agent. Steroid therapy (dexamethasone daily 40 mg) and IVIG (daily 35g) were initiated then these drug were stopped. CHOP-like regimen ( Etoposide 75 mg/m2, twice a week for two weeks then once a week until the seventh week) and Rituximab (375 mg/m2, once a week for 4 weeks) were initiated and continued for two weeks. Later on the patient died because of sepsis and multi-organ failure. Conclusions The multidisciplinary approach is very important. Physicians should be aware of HLH, because early recognition may prevent irreversible organ damage and subsequent death.4,5 In adults, HLH-associated mortality remains high, especially in patients with underlying malignancies. Collapsing glomerulopathy is the most commonly reported finding on renal biopsy. Renal prognosis appears to be poor with most patients remaining dialysis-dependent. The increased awareness of HLH, together with a more rapid diagnostic workup and new therapeutic approaches, will improve the prognosis of HLH in adults.

A247 VIRAL LOAD OF EPSTEIN-BARR VIRUS IN PEDIATRIC PATIENTS WITH NEW ONSET IBD AT DIAGNOSIS AND ON FOLLOW UP: AN OBSERVATIONAL STUDY

Abstract Background Patients with Inflammatory Bowel Disease are at increased risk for complications associated with Epstein-Barr Virus, such as uncontrolled infection, colitis mimicking IBD, and lymphoproliferative disease. These complications may be due to inherent immune dysfunction or effects of immunomodulating therapies used. We have previously identified that the seroprevalence of EBV in our cohort of IBD patients at time of diagnosis was 44.2%, with prevalence stratified by age as follows: 0 to &amp;lt;10 years 36%, 10 to &amp;lt;17 years 46%, and 17 + years 50%. Aims Our objective is to assess the risk of EBV reactivation in this population, to determine whether patients treated with immunomodulators should be more closely monitored for EBV viral load. Methods Retrospective chart review was done for all patients with new-onset IBD diagnosed at CHU Sainte-Justine over a two-year period, from Jan. 2016 to Dec. 2017. Serum from time of diagnosis was retrieved from the microbiology laboratory for patients with positive EBV serology, and quantitative PCR was performed to assess viral load at diagnosis. 47 of 53 seropositive patients had available serum at time of retrieval. EBV PCR was subsequently performed on serum drawn one to two years after start of immunosuppressants. Results 53 patients were EBV positive at time of diagnosis (EBNA/VCA IgG). Two patients were IgM positive, suggesting recent or active infection. The viral load as measured by quantitative PCR on serum drawn at diagnosis was negative in all retroactively tested patients. Of the two IgM-positive patients, one had known positive quantitative PCR at time of diagnosis. PCR previously tested in clinical follow-up of two seropositive, PCR-negative patients became positive at 7 and 17 months from diagnosis, suggesting viral reactivation. Both patients had received anti-TNF’s and systemic corticosteroids. Viral loads on follow-up are to be assessed for the rest of the cohort. Overall, therapies started within 6 months of diagnosis were similar in the seropositive and seronegative groups, the majority receiving some form of immunosuppression. Within the seropositive group: 66% received corticosteroids, 32.1% Infliximab, 5.7% Adalimumab, and 5.7% Azathioprine. Conclusions Only one patient had active EBV infection with positive PCR at time of diagnosis. All other patients had no sign of active infection based on retroactive PCR’s. While a majority of patients, regardless of EBV serology, receive immunomodulating agents, we currently do not routinely screen for seroconversion in seronegative patients, or for viral load in seropositive patients. We will be assessing viral loads after start of immunomodulation to better understand the potential impact of these agents on disease progression. Funding Agencies None

A57 VALPROATE INDUCED DRESS SYNDROME CAUSING MIXED LIVER INJURY

Abstract Background Drug reaction with eosinophilia and systemic symptoms (DRESS) syndrome occurs with 1 in every 1,000 to 10,000 drug exposures and has a mortality rate of up to 10%. It is one mechanism by which medications can induce liver injury with elevated liver enzymes seen in the majority of cases. In children, aromatic anticonvulsants are the drugs most commonly associated with DRESS syndrome. Valproate, a non-aromatic anti-epileptic, is not known to have a heightened risk of hypersensitivity syndromes and is often the anti-epileptic of choice in patients who develop hypersensitivity syndromes from other anti-epileptics. Valproate hepatotoxicity is normally caused by its inhibition of fatty acid transport and mitochondrial β-oxidation; vanishing bile duct syndrome is also reported. Aims We present a case, to the best of our knowledge the first in paediatrics, in which valproate causes DRESS syndrome and a secondary, predominantly cholestatic, liver injury. Methods Literature review and case report. Results A previously healthy 14-year-old girl was diagnosed with new-onset seizures and started on valproate. Three weeks later, she developed a pruritic exanthem. Despite discontinuing her valproate, the rash persisted and she developed fever and jaundice. She was admitted to the ICU at the Hospital for Sick Children with a diagnosis of DRESS syndrome (RegiSCAR DRESS score 7) for treatment with IV steroids. At admission, she had a skin eruption, fever, leukocytosis (22.86 x109/L), eosinophilia (1.03x109/L), atypical lymphocytes (1.62 x109/L), lymphadenopathy, and internal organ involvement (BiliC 138, GGT 501, INR 1.3, ALT 543, AST 370, Crt 109). Of note, her EBV PCR was positive. By discharge, her rash improved and kidney function normalized. Her cholestasis persisted, despite improvement in her transaminases and eosinophilia (BiliC 163, INR 1.0, GGT 338, ALT 506, AST 220, Eos 0.98x109/L). She was discharged home on an oral steroid wean, ursodeoxycholic acid, levocarnitine, and levetiracetam. Upon reaching a daily dose of 20mg of prednisone, her rash and pruritus worsened, she had lost 7kg, and she was readmitted for IV steroids. Her rash improved within 4 days and she was discharged on a slower steroid taper, with the addition of cholestyramine and insulin for steroid induced diabetes. At present (2 ½ months after diagnosis), she continues her oral steroids and has persistent liver injury and pruritus. Conclusions We report the first paediatric case of valproate induced DRESS syndrome causing a significant cholestatic presentation with otherwise preserved liver synthetic function. The prolonged cholestasis and pruritus may be a result of the natural course of DRESS syndrome, EBV reactivation/infection, or polypharmacy. Early recognition of DRESS syndrome as a mechanism of valproate induced liver injury in children is essential for earlier diagnosis and initiation of targeted therapy. Funding Agencies None