An audit of paroxysmal nocturnal haemoglobinuria (PNH) requesting at University Hospitals of North Midlands

Early identification of PNH, a rare life-threatening disease is essential to ensure appropriate management via the UK PNH service. Since testing for PNH is expensive (75.97 per test), we set out to assess the suitability of PNH requests against guidelines with the aim to feedback to colleagues and reduce unnecessary testing. To determine whether PNH requests at UHNM were in line with criteria in British Society for Haematology (BSH) guidelines. All patients over 18 years of age who had PNH testing for the first time and those who had repeat testing for monitoring between 01/04/2019 and 31/03/2020 were included. Patients were selected from electronic records of PNH sample receipt to laboratories. Hospital records were reviewed for clinical details and investigation Results. 82 requests including 79 individual patients were audited. 57% were male and 43% were female. Median age was 56 years. 97.6% of PNH tests were requested by a haematologist whilst only 2.5% requests were done by non-haematology clinicians. 52.4% requests were in keeping with BSH recommendations, whilst 47.6% tests did not meet criteria for testing. All patients tested outside of guideline recommendations were negative. Of the reasonable requests, only 23.3% (10) were positive. Of the PNH positive patients, 8 patients were known to have a PNH clone with aplastic anaemia;one patient had a hypoplastic bone marrow and a known PNH clone whilst only one patient with cytopenia had a new positivity for PNH. The frequency monitoring for aplastic anaemia and a PNH clone was 100% concordant with BSH recommendations. With appropriate testing, only one new patient was identified. Our audit has limitations. We have not been able to assess whether any patients outside of those monitored for PNH in aplastic anaemia have been overlooked for testing. Also, the time period includes the COVID-19 pandemic so our findings may not reflect usual practice. New BSH guidelines for thrombophilia testing were published in 2022 and recommend testing for PNH in patients with thrombosis at unusual sites and abnormal haematological parameters and for patients with arterial thrombosis and abnormal blood parameters. This will likely limit excess tests although the sensitivity and specificity of such an approach has not been formally evaluated. In a finite health system, it is our responsibility to rationalise investigations. The cost of a year's testing was 6229.54 and that of inappropriate testing was 2962.83. As a department, we could, therefore, save 2962.83.

Primary immunodeficiency associated with a SAMD9L mutation

Background: SAMD9L is a tumor suppressor involved in regulating the proliferation and maturation of cells, particularly those derived from the bone marrow, and appears to play an important role in cerebellar function. It can be activated in hematopoietic stem cells by type I and type II interferons. It has been hypothesized to act as a critical antiviral gatekeeper regulating interferon dependent demand driven hematopoiesis. Gain of function mutations can present with an immunodeficiency due to transient severe cytopenias during viral infection. Case presentation: We report a 3-year-old boy born full term with a history of severe thrombocytopenia requiring transfusions, developmental delay, ataxia, seizure disorder, and recurrent severe respiratory viral infections. His infectious history was significant for respiratory syncytial virus with shock requiring extracorporeal membrane oxygenation complicated by cerebral infarction and a group A streptococcus empyema, osteomyelitis requiring a left below the knee amputation, and infections with rhinovirus, COVID-19, and parainfluenza requiring hospitalizations for respiratory support. Initial immunologic evaluation was done during his hospitalization for parainfluenza. His full T cell subsets was significant for lymphopenia across all cell lines with CD3 934/microL, CD4 653/microL, CD8 227/microL, CD19 76/microL, and CD1656 61/microL. His mitogen stimulation assay to phytohemagglutinin and pokeweed was normal. Immunoglobulin panel showed a mildly decreased IgM of 25 mg/dL, but normal IgA and IgG. Vaccine titers demonstrated protective titers to 12/22 pneumococcus serotypes, varicella, diphtheria, mumps, rubella, and rubeola. Repeat full T cell subsets 6 weeks later revealed marked improvement in lymphocyte counts with CD3 3083/microL, CD4 2101/microL, CD8 839/microL, CD19 225/microL, and CD1656/microL. A primary immunodeficiency genetic panel was ordered and positive for a heterozygous SAMD9L c.1549T>C (p.Trp517Arg) mutation classified as a variant of unknown significance. Discussion(s): This patient's history of severe viral infections, ataxia, thrombocytopenia, and severe transient lymphopenia during infection is suggestive of a SAM9DL gain of function mutation. Protein modeling done by the laboratory suggests this missense mutation would affect protein structure. The mutation found has been observed in individuals with thrombocytopenia. This case highlights the importance of immunophenotyping both during acute illness and once recovered.Copyright © 2023 Elsevier Inc.

Characteristics and Clinical Implications of Anti-IFN-alpha cytokine antibodies in partial Recombinase Activating Gene Deficiency patients before and during the COVID-19 Pandemic

Introduction: With the onset of the COVID-19 pandemic, there was increased attention on anti- IFN-alpha autoantibodies and its correlation with severe clinical outcomes in a large group of patients. However, this correlation has not been extensively investigated in patients with partial Recombinase Activating Gene Deficiency (pRD) who are known to have increased prevalence of anti- IFN-alpha autoantibodies. Therefore, there is a need to assess the presence of anti- IFN-alpha antibodies in pRD patients before and after the COVID-19 pandemic and explore the relationship between anti- IFN-alpha antibody presence and clinical outcomes. Method(s): Sera was collected from the whole blood after informed consent and Enzyme-Linked Immunosorbent Assay was conducted to confirm the presence of IgG-specific anti- IFN-alpha autoantibodies. Positive samples were determined as OD values above 3 standard deviations of the healthy donor OD mean. Result(s): Our cohort included both adult (n = 13) and pediatric (n = 9) patients with variants in RAG1 and RAG2. Eleven patients (50%) out of the 22 showed elevated anti- IFN-alpha autoantibodies levels. Five patients (23%) were defined as low positive for anti- IFN-alpha autoantibodies, and 6 patients had no autoantibody titers. Of the 22 patients, 16 were symptomatic with infectious and non-infectious complications including recurrent viral and/or bacterial infections, autoimmune cytopenias, and lymphoproliferation. Ten (63%) of the symptomatic patients demonstrated high anti-IFN-alpha autoantibodies titers. Of the 11 patients with no or low neutralizing anti- IFN-alpha autoantibodies levels, 5 were asymptomatic. In temporal comparison, 16 samples were collected pre-COVID-19 pandemic;8 samples were collected during the pandemic, 2 of which belonged to patients with samples collected before and during the pandemic. In the pre-pandemic cohort, 66% had anti- IFN-alpha autoantibodies. Conversely, during the COVID-19 pandemic, 89% had anti- IFN-alpha autoantibodies. Of note, one patient who had neutralizing anti- IFN-alpha autoantibodies remained positive both before and during the pandemic despite HSCT. Patient also had a SARS-CoV-2 infection in summer of 2022 with a mild clinical course. Conclusions & Next Steps: We observed persistence of anti-IFN-alpha autoantibodies in our cohort post-pandemic and even post-HSCT. It is unclear whether the presence of anti-cytokine antibodies are risk factor for severe COVID-19.Copyright © 2023 Elsevier Inc.

BTKi-treated CLL patients have Favourable survival outcomes but concerning levels of second malignancies and COVID-related mortality

Introduction: Bruton's tyrosine kinase inhibitors (BTKi) are used extensively within the NHS to treat specific B-cell malignancies with patients stopping BTKi usually due to adverse events or disease progression. The objective of this study was to analyse effectiveness of BTKi therapy for chronic lymphocytic leukaemia (CLL) at our centre compared to previously published real-world data from the UKCLL Forum (Follows et al, Blood 2019). In addition, we investigated treatment-related adverse events (AE) and second malignancies. Method(s): This is a single-centre retrospective study of 112 CLL patients treated with a BTKi for a minimum of 4 weeks between 2014 and 2022 (ibrutinib n = 71, acalabrutinib n = 38, zanubrutinib n = 3). Treatment was first line (n = 39), second line (n = 44) and 3+ line (n = 29). Patient demographics, duration of BTKi therapy, Aes, discontinuation reasons and second malignancies were collected. Aes were compared with a parallel cohort of 53 non-CLL BTKi-treated patients. Result(s): Median age starting treatment was 73 years, and 71% were male. Primary outcomes were discontinuation-free survival (DFS) and overall survival (OS). With a median follow-up of 3.90 years, the median DFS was 4.18 years (95% CI: 3.52-4.91) with a median OS of 6.35 years (95% CI: 5.52-NA). These compare favourably with previous UKCLL forum data (median DFS = 2.79 years;median OS = 4.66 years), although our patients were more likely to receive BTKi earlier in treatment (3rd line or beyond: 26% of our patients vs. 78% in the UKCLL Forum). The most common Aes included bleeding, cytopenia, infection, cardiac events and mouth ulcers, with 21% stopping BTKi for CLL due to Aes whilst 15% of non-CLL BTKi patients stopped due to an AE. Second malignancies were reported in 49% of CLL patients, yet only 34% of non-CLL patients. Among patients with a confirmed cause of death, infection was the most common cause (39%), followed by CLL (33%), then second malignancy (18%). Of the 31 deaths in 2020 and 2021, 7/31 (23%) were due to, or in association with COVID-19 infection. No COVID-19 deaths were associated with BTKi in non-CLL patients. Conclusion(s): We demonstrate a favourable real-world DFS and OS for BTKi-treated CLL patients although a high number of patients still stop BTKi due to Aes. The very high incidence of second malignancies for all BTKi-treated patients and COVID-19 and infection-related deaths for CLL patients is concerning. As CLL is known to associate with high levels of second cancers, it remains unclear whether BTKi use increases this risk further.

Real-World Single-Centre Experience of Jak Inhibitor Use in Ra

Background/Aims The use of Janus Kinase Inhibitors (JAKi) has been gradually increasing overtime in the management of rheumatoid arthritis (RA) and other inflammatory arthritis and these appeal to patients. being oral agents. Nevertheless, rheumatologists have become cautious about their use since recent trials have shown safety concerns about VTEs, MACE and malignancies. Methods We decided to study use of JAKi at our centre in Princess of Wales Hospital Bridgend. The aim was to assess whether appropriate patients were selected (considering cautions about MACE, VTEs and malignancies). We also wanted to see whether all patients had required pretreatment safety testing and post-treatment monitoring performed. Results These were 70 patients;59 were females and 11 were males. All of them were diagnosed as RA. Average age was 61.1 years (20-85), average duration of disease 129.9 months (16-340) and average duration of treatment was 58.1 weeks. The most common JAKi being used was baricitinib (84%) followed by tofacitinib (12%) and upadacitinib (4%). 50% patient were on concomitant csDMARDs among whom two-thirds were on methotrexate. Looking at previous biologic use, 9 patients were biologic naive, 22 had one biologic, 15 had two biologics used in the past. All patients were appropriately selected (severe RA and no significant risk factors for MACE, VTEs and malignancies). All patients had pre-treatment Hepatitis B, Hepatitis C, latent TB, FBC and LFTs checked. All patients had FBC and LFTs monitored post treatment. No patient developed VTE, MACE or cancer on treatment. 84.2% patients had lipids tested before starting JAKi. 22.8% patients had abnormal lipids before Rx initiation and 62.5% of these were on lipid lowering Rx. All patients had lipids tested post treatment, but the timing was quite variable and only 62.5% of patients had lipids tested on the recommended time. There were 2 deaths recorded in this cohort. One of those was an 80-year-old RA patient on baricitinib 2mg OD, who died due to chest infection on the background of ILD. He was not on steroids or csDMARDs. The second patient was 63 years' old (on baricitinib 4mg OD), and died due to respiratory sepsis, and was also on azathioprine. She had RA with advanced ILD. The reasons for discontinuing JAKi were inefficacy (46%), side effects (39%) and both inefficacy and side effects (15%). 41.4%of patient experienced side effects due to JAKi. These included infection 28%, deranged lipids 17%, cytopenia 14%, deranged LFTs 14%, GI side effects 10%, skin rash 7% and varicella zoster 3%. Conclusion There has been steady increase in the use of tsDMARDs for RA and other rheumatic conditions. Due to short half-life, these drugs became a popular choice during COVID-19 pandemic but on the other hand safety monitoring became extremely challenging during this time.

TCR Alpha Beta and CD19+ Depleted Haploidentical Peripheral Stem Cell Transplantation Using Reduced Toxicity Conditioning Regimen with Pharmacokinetics Guided Busulfan, Fludarabine, Thiotepa and Thymoglobulin Offers Durable Donor Engraftment in Children with Primary Immune Deficiency Disorders

Background: Allogeneic hematopoietic cell transplantation (HCT) with ex vivo T cell receptor (TCR) alphabeta+ T cell and CD19+ B cell depletion is an effective approach for children with primary immune deficiency disorders (PIDD) as it combines advantages of high CD34+ cell dose facilitating rapid engraftment with low risk of Graft Versus Host Disease (GVHD). The ideal pre-conditioning regimen that facilitates robust donor engraftment without increasing risk of transplant related mortality has not been well defined with this approach. Method(s): We report the outcomes of 4 pediatric subjects: Chronic Granulomatous Disease (CGD) (2), Wiskott Aldrich Syndrome (WAS) (1), and RAC2 deficient Severe Combined Immunodeficiency (1) who underwent haploidentical HCT with TCRalphabeta+ T cell/CD19+ depletion at Johns Hopkins All Children's Hospital/Moffitt Cancer Center from 2020-2022 (NCT04414046). Pre-conditioning regimen consisted of distal thymoglobulin (7.5 mg/kg), fludarabine (175 mg/m2), thiotepa (10 mg/kg) and pharmacokinetic guided busulfan targeting a cumulative area under curve (cAUC) (65-75 mgxhr/L). Rituximab (200 mg/m2) was administered on day +1. Result(s): The median age at HCT was 51 months (range 10-163 months). All patients received mobilized peripheral blood stem cells from HLA- haploidentical donors (paternal=1, maternal=1 sibling=2). Median busulfan cAUC for all patients was 69 mgxhr/L (range 65-76). Median CD34 and TCR alphabeta T cell dose was 9.13x106 cells/kg (range 7.0-18.9x106) and 0.7x105 cells/kg (range 0.09-1.0x105). Median times to neutrophil and platelet engraftment were 11 days (9-12) and 11 days (range 8-15), respectively. All 4 patients are alive with median follow-up of 19.5 months (range 7-24). One patient developed late VOD without organ dysfunction that resolved with defibrotide. At last follow up, peripheral T and myeloid chimerisms exceeded 90% in all 4 patients. Average time to CD4 recovery (> 200x106/L) was 142 days. Pre-existing inflammatory bowel disease in CGD (n=1) and WAS (n=1) patients resolved immediately following transplant. There was no graft failure, and none developed Grade III-IV acute or extensive chronic GVHD. Patient with WAS developed recurrent autoimmune cytopenias requiring corticosteroids, rituximab, sirolimus and daratumumab, and ultimately resolved. Viral reactivations included EBV (n= 1), adeno (n= 1), HHV6 (n= 2), BK (n=1), norovirus (n=1), and late HSV (n=1), all responded to antivirals without disease. All patients acquired SARS-Cov-2 after transplant and recovered without sequelae. Conclusion(s): TCR alphabeta+ and CD19+ depleted haploidentical transplantation using a reduced toxicity conditioning regimen with pharmacokinetic guided busulfan, fludarabine, thiotepa and thymoglobulin is well-tolerated in young children with PIDD that results in rapid, durable engraftment with low likelihood of GVHD and graft rejection.Copyright © 2023 American Society for Transplantation and Cellular Therapy

Outcomes for Anti-CD19+ Chimeric Antigen Receptor T (CAR-T) Cell Therapy for Non Hodgkin Lymphoma (NHL) across a Large Community Transplant and Cellular Therapy Network

Background: The Sarah Cannon Transplant and Cellular Therapy Network (SCTCTN), which offers community access to transplant and cell therapy, implemented a coordinated approach to deliver CAR-T therapy through 5 programs. We conducted a retrospective review of clinical outcomes after FDA-approved anti-CD19+ CAR-T in B-cell NHL. Method(s): All patients referred for evaluation within SCTCTN were tracked in our prospective registry (Stafa-CT). We identified 110 patients who received FDA-approved anti-CD19+ CAR-T for NHL within the network between 12/10/2018 and 3/7/2022. All patients received care through standardized eligibility criteria, process, care pathways, toxicity management protocols, and a single quality plan. Result(s): The median age at referral was 60 years (range 23-82), 63% were male, the referral indication was diffuse large B-cell lymphoma (70%), mantle cell lymphoma (7%), follicular lymphoma (15%), or other B-NHL (8%). 35% had received a prior autologous transplant. The median time from referral to infusion was 143 days (range 89- 224), and from collection to infusion was 32 days. The infusion year was 2018 (1), 2019 (20), 2020 (31), 2021 (48), 2022 (10). The CAR-T cell products were Axi-cel (70), Tisa-cel (27), Brexu-cel (9), and Liso-cel (4). 16 patients (15%) were infused as outpatient, of which 10 patients were subsequently hospitalized at a median of 8 days (range 1-26) after infusion. Of the 94 patients (85%) infused as inpatient, the median length of stay was 15 days (range 6 to 85). Cytokine release syndrome (CRS) was observed in 78% with a median maximum grade 1. Maximum grade CRS was none, grade 1, grade 2, grade 3, grade 4, grade 5 in 22%, 36%, 32%, 7%, 2 % and <1%, respectively. The median times to onset and resolution of symptoms were day 3 and 8, respectively. Tocilizumab was administered to 39% for a median of 2 doses. Neurotoxicity was observed in 55% with a median maximum grade 1. Maximum grade neurotoxicity was none, grade 1, grade 2, grade 3, grade 4, grade 5 in 45%, 19%, 13%, 18%, 4 % and 0%, respectively. The median times to onset and resolution of symptoms were day 7 and 13, respectively. Neutropenia (<0.5/ muL) and thrombocytopenia (<20K/muL) at day 30 were reported in 11% and 12%, respectively. 18% required ICU stay. 37 deaths (34%) were reported from disease progression (23), infections (7, including 5 from COVID), CRS (2) and other causes (5).(Figure Presented) Conclusion(s): Administration of anti-CD19+ CAR-T is feasible in specialized community hospitals with outcomes similar to registrational clinical trials. Outpatient administration is feasible in selected patients, but subsequent hospitalization needs to be anticipated. CRS, neurotoxicity, cytopenias and infection remain challenges, while disease progression was the commonest cause of deathCopyright © 2023 American Society for Transplantation and Cellular Therapy

Examining the Role of Hematological Parameters among COVID-19 Patients: An Analytical Study

Aim: This study was aimed at examining the role of hematological parameters among COVID-19 patients in Bihar. Method(s): The study was conducted at Bhagwan Mahavir Institute of medical science, pawapuri, Bihar, India for 7 months to compare hematological parameters of red blood cells (RBCs), platelets, and white blood cells (WBCs) among patients with and without COVID-19 diagnosis. In this study, 500 patients were recruited, a study group of 250 patients testing positive and a control group of 250 testing negative. Result(s): The result showed that 160 (64%) of COVID-19 patients were male and 90 (36%) were female, while 150 (60%) of non-COVID-19 patients were male and 100 (40%) were female. The age range of COVID-19 patients was 20-90 years old;150 of these (60%) between 30 and 60 years old, 75 (30%) over 60, and the remaining 25 (10%) below 30. The non-COVID-19 patients' age range was 20-88;150 of these (60%) between 30 and 60, 55 (22%) over 60, and the remaining 45 (18%) below 30. Regarding the clinical information of COVID-19 patients, 79 of them (31.6%) were attending the hospital in critical status, 54 (21.6%) with mild symptoms, 50 (20%) asymptomatic, 52 (20.8%) with moderate symptoms, and 25 (10%) with severe symptoms. Regarding the COVID-19 patients' situation during the study period, 175 (70%) recovered and were discharged from the hospital, 25 (10%) were still ICU patients at the end of the study period, 15 (6%) were isolated in hospital wards, and 35 (14%) unfortunately passed away. Conclusion(s): Our study results indicate that mild anemia associated with leukopenia may have diagnostic value for COVID-19. Careful assessment of hematological parameters, at baseline and throughout the disease path, will assist physicians in formulating personalized approaches to treatment and promptly offer intensive care to those in greater need.Copyright © 2023, Dr Yashwant Research Labs Pvt Ltd. All rights reserved.

X-Linked Inhibitor of Apoptosis (XIAP) deficiency presenting as hemophagocytic lymphohistiocytosis triggered by drug allergy

X-linked inhibitor of apoptosis (XIAP) deficiency is a primary immunodeficiency associated with recurrent hemophagocytic lymphohistiocytosis (HLH) episodes. The clinical phenotypes of XIAP deficiency vary, ranging from splenomegaly to life-threatening inflammation. We report a case of XIAP deficiency with unusual late-onset HLH presentation likely triggered by a drug allergy. A previously healthy adolescent boy presented to the hospital with fever and rash seven days after starting antibiotics for a neck abscess. Laboratory evaluation demonstrated cytopenias, elevated liver enzymes, and increased inflammatory markers. Initially, antibiotics were discontinued due to concern for drug rash. He continued to deteriorate clinically and became hypotensive. Additional testing revealed decreased NK cell function, as well as elevated ferritin, triglycerides, and soluble IL-2 receptor. SLAM-Associated Protein (SAP) and XIAP evaluation by flow cytometry demonstrated decreased XIAP expression. Subsequently, genetic testing revealed a known pathogenic mutation in BIRC4 (c.421_422del), confirming the diagnosis of XIAP deficiency.Copyright © 2023

The role of lymphocyte transformation tests (LTT) in suspected severe delayed hypersensitivity reactions following COVID-19 vaccination.

Coronavirus (COVID-19) vaccines have proved to be effective in the pandemic response but can cause adverse events such as delayed hypersensitivity reactions (DHRs). Delayed-reading intradermal tests (IDT) to vaccines are limited by false-positive results and may reflect a cell-mediated rather than IgE-mediated immune response. Lymphocyte transformation test (LTT), which has been utilized in the diagnosis of drug allergy, may be helpful in suspected COVID-19 vaccine and/or its excipient-related DHRs. To investigate the use of LTT in two suspected cases of COVID-19 vaccine-induced DHRs, two patients with suspected DHRs to COVID-19 vaccination were tested by delayed-reading IDT and LTT against vaccines and their excipients. A 47-year-old man developed acute mixed-pattern hepatitis after the second dose of ChAdOx1 vaccine. LTT performed at 2 months post-vaccination revealed reactivity to the ChAdOx1 vaccine, polysorbate 80 and mildly to PEG 2050 but not BNT162b2 vaccine. Delayed-reading IDT returned negative to both vaccines and excipients. He tolerated BNT162b2 vaccination with no adverse events. A 36-year-old woman presented with subacute morbilliform eruption and hepatitis after the first dose of BNT162b2 vaccine. LTT performed 3 months later revealed reactivity to the BNT162b2 but not PEG 2050. Repeat LTT following subsequent natural Severe Acute Respiratory Coronavirus-2 (SARS-CoV-2) infection revealed reactivity to ChAdOx1 and NVX-CoV2373 vaccines but not polysorbate 80. Delayed-reading IDT remained negative. She proceeded with NVX-CoV2373 vaccination with no symptom recurrence. LTT may be a useful tool in suspected COVID-19 vaccine-related DHRs. Further evaluation with a larger patient cohort is required.

GATA2 deficiency presenting as fever and rash

Case Report: Our patient is an 8-year-old Caucasian female with a history of choanal atresia, first degree heart block, recurrent urinary tract infections, and recent COVID-19 infection, who initially presented with an episode of syncope and vomiting. By history, she had two weeks of daily fever and an intermittent nonspecific rash. She was diagnosed with a UTI 5 days prior to presentation but had not defervesced despite treatment. Shewas initially found to be in shock with tachycardia and poor perfusion and was treated with fluid resuscitation, antipyretics, and empiric antibiotics. Labs were significant for leukopenia, elevated inflammatory markers, lactic acidosis, coagulopathy, and mildly elevated troponin. Chest x-ray showed abnormal but non-specific widespread infiltrates. She was initially treated with IVIG and pulse steroids for a working diagnosis of MIS-C, however she did not improve and a more extensive infectious, oncologic, and rheumatologic work-up was performed. Her workup revealed a disseminated Mycobacterium abscessus infection. Bone marrow biopsy revealed myelodysplasia with monosomy 7. Her buccal swab testing revealed a heterozygous germline mutation in the GATA2 gene, a variant that is predicted to cause loss of normal protein function. She is presently on multidrug regimen for her mycobacterial infection. Her myelodysplasia evolved into an acute leukemia, and she is undergoing chemotherapy for that at this time. Discussion(s): GATA2 deficiency, first identified in 2011, is a rare immune disorder resulting in a wide variety of clinical presentations. It is caused by a germline mutation of the GATA2 gene that disrupts blood cell differentiation, resulting in decreased or absent monocytes, B cells, NK cells, and dendritic cells1. This case presented multiple challenges due to the broad range of differential diagnoses. This patient was ultimately diagnosed with myelodysplastic syndrome associated with monosomy 7 and GATA2 deficiency, confirmed by FISH testing. Due to the presentation and lab derangements this patient had, there was a delay in targeted treatment while managing her cytopenias and presumed pulmonary infection. GATA2 deficiency carries a high risk of progression from myelodysplastic syndrome to acute myelogenous leukemia. The best long-term treatment for GATA2 deficiency is hematopoietic stem cell transplant, which is the ultimate goal for our patient. Copyright © 2023 Southern Society for Clinical Investigation.

Severe Covid-19 Associated Hemophagocytic Lymphohistiocytosis : A Comparative Study with Hemophagocytic Lymphohistiocytosis Secondary to Causes Other Than Covid-19

Introduction: Severe COVID-19 infection in a subset of patients is associated with hyperinflammation similar to hemophagocytic lymphohistiocytosis (HLH) however it may not fulfill the required diagnostic criteria (HLH 2004 criteria or H score). We compared clinical, laboratory parameters, bone marrow findings and disease outcome of severe COVID-19 infection related HLH with HLH secondary to causes other than severe COVID-19 to describe features specific to severe COVID-19 associated HLH and limitations of currently available diagnostic criteria of HLH in context to severe COVID -19 infection induced hyperinflammation. Method(s): We analyzed 69 patients diagnosed as HLH of which 47 had severe COVID-19 and 22 had HLH secondary to causes other than COVID-19. Clinical, hematological and biochemical parameters were compared using Mann-Whitney U test. Bone marrow examination (BME) was done in all for presence of hemophagocytosis. Immunohistochemical staining for CD68 and CD163 were done for identification of histiocytes. Occurrence of COVID-19 related HLH was taken as the dependent variable to determine predictors of COVID-19 HLH. Result(s): Organomegaly was seen in only 4.3% (2/47) cases with COVID-19 related HLH as compared to 54.5% (12/22) with non-COVID HLH (p< 0.001). Amongst the quantitative variables, a significant difference in COVID-19 related and non-COVID-19 related HLH was found in the following parameters: Age (p< 0.001), Triglyceride (p=0.009), Fibrinogen (p< 0.001), Ferritin (p< 0.001), Hemoglobin (p< 0.001), Total leukocyte count (p=0.003), Absolute neutrophil count (p< 0.001), Neutrophil lymphocyte ratio (p< 0.001) and H score (p< 0.001). BME of all patients showed presence of hemophagocytes. Only 6.4% (3/47) cases with COVID related HLH had 5/8 HLH 2004 criteria as compared to 63.6% (14/22) cases with non-COVID related HLH (p< 0.001). H-Score >=169 was also significantly less common in COVID HLH as compared to non-COVID HLH (40.42% vs 86.36%, p=0.001). Conclusion(s): Organomegaly, cytopenias, hypofibrinogenemia and hypertriglyceridemia which are part of HLH diagnostic criteria are rare in severe COVID-19 making it difficult to diagnose. Demonstration of hemophagocytes in bone marrow should be recommended in suspicious cases for initiation of early immunosuppressive therapy. (Figure Presented).

Cytopenias in Hospitalized Patients of Covid-19 during First and Other Waves

Introduction: COVID -19 emerged in December 2019 in china and till date this virus has mutated into different strains which are somewhat different in their presentation and severity. Pakistan has faced five waves of COVID-19 till date. This study was aimed to compare Haematological parameters and outcomes of hospitalized patients during First and other waves (second and third) of COVID-19. Method(s): Demographic, Clinical and laboratory data as well as outcome of total 202 COVID-19 PCR positive patients admitted in Fatima memorial Hospital, Lahore, Pakistan was collected during first and other waves. Data of First wave is from May 2020 to July 2020, second wave from early November to Mid December 2020 and third wave from Mid march to June 2021. We compared the data on basis of SII as well hematological parameters of First Wave were compared with other waves (second & third) of COVID-19. Result(s): There were 54(26.7%) patients with SII< 600 and 148(73.2%) patients with SII>600. Neutrophilia (p-< 0.001), leukocytosis (p-< 0.001), lymphocytopenia(p-< 0.001), Raised NLR(p-< 0.001), PLR(p-< 0.001) and MLR(p-< 0.001) had direct association with SII. Raised SII was directly related to increased requirement of ventilator support (p-0.2) and Mortality (p-0.001). There were total 90(44.5%) patients in first wave and 112(55.4%) patients in other waves of COVID-19. More Females (33.3% Vs 57.1%) than Males (66.7% Vs 42.9%) were infected during other waves (p- 0.001). Anemia (27.8% vs 37.5%) (p-0.09), leucopenia (1.1% vs 8.9%) (p-< 0.001), Lymphocytopenia (20% Vs 39.3%) (p-0.003) and thrombocytopenia (3.3% Vs 25%) (p-< 0.001) were more prevalent in other waves of COVID-19 when compared to first wave. Requirement of Ventilator Support (7.7% vs 22%) and mortality (10% vs 33.9%) were also increased in other waves of COVID-19 Conclusion(s): Systemic immune Inflammation Index (SII) is a good predicting tool in COVID -19 patients for prognosis and is directly associated with severity of disease as well as outcome. Patients with cytopenias like anemia, leucopenia, lymphocytopenia thrombocytopenia, requirement of Ventilator Support and mortality were noted to be more in other waves of COVID-19. Haematological parameters like ANC, AMC NLR, PLR, MLR and SII show no significant difference between first and subsequent waves of COVID-19.

Blood Count Abnormalities and Covid-19 in Pediatric Patients

Introduction: Coronavirus disease 2019 (COVID-19) is a systemic infection with a significant impact on the hematopoietic system. In pediatrics, COVID-19 is known to cause less severe disease course and transient hematologic abnormalities, including leukopenia, lymphopenia, anemia and thrombocytosis or thrombocytopenia Methods: This retrospective study conducter in the hematology departement at IBN ROCHD hospital university center over a period of 2 years between January 2020 and January 2022 involing patients with COVID-19 infection according to WHO recommendations in whom the labs showed the cytologics abnormalities. Result(s): This study included 12 patients whom had anormal blood count The average age was 7.2 ( 2 ans - 16 ans) with sex ratio M/F = 0.5. Three patients had cytopenia (25%) tow patients had anemia and one with thrombopenia Labs showed bicytopenia with anemia (10.5 g/dl) and thrombocytopenia ( 100.000/mm3) in tow patients (16%) and one patient had anemia and neutropenia Tow patients (16%) had lymphopenia . Labs showed anemia and hyperleukostasis with PNN in three patients( 25%). One patient had a severe lymphopenia ( 60/ mm3) and thrombocytopenia ( 31.000 /mm3). One patient presented with thrombocytosis ( 556.000/mm3) Conclusion(s): Lymphopenia may be considered as a cardinal laboratory finding. In the pediatric population the different cytological abnormalities may have prognostic value Coronavirus disease 2019 (COVID-19) in determining sever cases. Hence the interest in conducting studies to correlate the clinical course of the infection with the results of biological examinations.

COVID-19 in cancer patients - higher inflammatory response in patients with active cancer

Background: Active cancer has been identified as an independent risk factor for severity and mortality in COVID-19. However, direct comparisons of SARS-CoV-2 infected patients (pts) with active and non-active cancers remain scarce. Method(s): We retrospectively analyzed a cohort of pts with cancer with confirmed SARS-CoV-2 infection, enrolled 03/16/2020 - 07/31/2021. Data on demographics, cancer and laboratory findings were collected. Descriptive and subsequent regression analysis was performed. Endpoints were progression to severe COVID-19 and infection-associated mortality. Result(s): In total, 987 pts with cancer (510 active vs 477 non-active) were included in our analysis. Majority was male and > 55 years, with a higher number of elderly pts with non-active cancer. CCI was 4.75 vs 3.85 in pts with active and non-active cancer (p<0.001). Localized solid tumors were reported in 38 vs 79% (p<0.001), metastasized in 37.5 vs 5.5% (p<0.001) and hematological diseases in 37.5 vs 19.5% (p<0.001) pts with active and non-active cancer, respectively. At virus detection, majority of pts showed mild to moderate symptoms, while deterioration to severe COVID-19 was slightly more common in pts with active cancer (19% vs 16%;p=0.284). COVID-19 related mortality was significantly higher in pts with active cancer (24% vs 17.5%, p<0.001). In line, severe cytopenia and an increase of inflammatory markers were common findings in pts with active cancer at baseline, particularly in those who developed severe infection or died. Multivariate analysis revealed that ferritin (14.24 [2.1-96], p=0.006) and CRP (2.85 [1.02-8.02], p=0.046) were associated with severe COVID-19 and infection-related mortality. In pts with non-active cancer, association was seen for ferritin only (4.1 [1.51-11.17], p=0.006). Conclusion(s): Comparing pts with active and non-active cancer, mortality rate was significantly higher in pts with active cancer. Also inflammatory markers were significantly increased assuming higher levels of inflammation may play a role in adverse outcome of COVID-19 in pts with active cancer.

Ctla4 Variant of Uncertain Significance in a Patient with a History of Cytopenias

Introduction: CTLA-4 haploinsufficiency is caused by heterozygous mutations in CTLA4, a negative regulator of immune responses. Hypogammaglobulinemia, infections, and autoimmune cytopenias can be seen. Here, we describe a patient with history of ITP who presented with neutropenic fever, lymphopenia, and hypogammaglobulinemia in the setting of COVID-19 with a VUS in CTLA4. Case Description: The patient is a 24-year-old female diagnosed with ITP at 10-years-old, initially treated with IVIG/steroids, then on mycophenolate for 6-years. 5-years later, she had a relapse of ITP in the setting of viral illness, requiring steroids/IVIG/rituximab. She developed neutropenic fever, unresponsive to GCSF, but responsive to cyclosporine and was noted to have a LGL clone. At 24-years-old, she was admitted with neutropenic fever (ANC 0);adenovirus, parainfluenza-virus, and SARS-CoV-2 were positive. Immunology was consulted due to hypogammaglobulinemia (IgG 140, IgA 7, IgM <5 mg/dL). Prior genetic testing identified a missense VUS in CTLA4 c.370A>C (P.Thr124Pro), shown to affect CTLA4 function and observed in individuals with clinical features of CTLA-4 haploinsufficiency. Lymphopenia, absent lymphocyte antigen responses, and impaired vaccine immunity were noted. ANC improved with GCSF. The patient was discharged with immunology follow-up for consideration of abatacept. Discussion(s): This case highlights the importance of considering VUS in the diagnosis and treatment of primary immunodeficiency. Our patient had a history of recurrent ITP, neutropenia, and LGL clone, all likely manifestations of CTLA-4 haploinsufficiency. Subsequent recognition of hypogammaglobulinemia and lymphopenia in the setting of neutropenia supported the diagnosis. Abatacept replaces the missing CTLA4-protein and should be considered in patients with CTLA-4 haploinsufficiency presenting with cytopenias. Copyright © 2022

Role of copper levels in patients with myelodysplastic syndromes

Background: Patients with inadequate amounts of copper often present with cytopenias and exhibit dysplasia on bone marrow, mimicking myelodysplastic syndrome (MDS) and pose diagnostic difficulties. Method(s): This cross-sectional observational study was performed from January 2020 to June 2021. Patients diagnosed with MDS were included in the study and serum copper levels were measured by the Inductively Coupled Plasma Mass Spectrometry (ICPMS) method. Copper supplementation with intravenous copper chloride 2.5mg daily for the first two weeks, followed by oral 3mg copper sulfate thrice daily for the next three months, was given for copper-deficient patients. Response assessment was performed with repeat hemogram and serum copper levels. Result(s): A total of 57 patients were diagnosed with MDS, of these, 33 (57.89%) were males and 24 (42.10%) were females. The mean age was 54.3+/-14.6 years (13-81). The distribution of patients in different types of MDS was MDS-SLD in 15, MDS-MLD in 18, MDS-EB1 in 7, MDS-EB2 in 8, and MDS-U in 9 patients. Anemia was seen in (87.71%) of patients, with mean hemoglobin 7.6+/-2.1g/dL (4.6-14.5g/dL). Neutropenia was seen in 31 (54.38%) with a mean absolute neutrophil count(ANC) of 2073+/-2139/muL (211-10,952/muL). Thirty seven (64.91%) patients had thrombocytopenia with a mean platelet count of 1,05,298+/-1,21,769/muL (9,000-6,74,000/muL). The mean serum copper levels were 146.69+/-42.36mug/dL (54.2-254.0mug/dL). Only three (5.26%) patients out of 57 were found to have copper deficiency. All three patients with low copper levels were found to have anemia, thrombocytopenia, and mildly raised serum erythropoietin levels. All three patients had dyserythropoiesis on bone marrow examination, and only one patient each had cytoplasmic vacuolations in erythroid precursors and dysmegakaryopoiesis. Among the three patients with copper deficiency, two patients had significant improvement in cytopenias after copper supplementation, and one had lost follow-up due to COVID-19. Conclusion(s): This study is the first from India to evaluate the role of copper in patients presenting with predominantly hematological manifestations. For patients presenting with cytopenias or marrow dysplasia resembling MDS, copper deficiency should be considered in the differential diagnosis. Legal entity responsible for the study: The authors. Funding(s): Has not received any funding. Disclosure: All authors have declared no conflicts of interest. Copyright © 2022

IgA Vasculitis After COVID-19 Vaccination: A Rare but Significant Complication

Introduction: A 24 year old male presented with rash, gastrointestinal bleeding and nephrotic syndrome one week after first COVID vaccination. Renal biopsy revealed crescentic IgA nephritis. He was treated steroids and responded clinically but continues to have proteinuria. Case Description: A 24 year old male with history of cerebral palsy presented with a vasculitic rash, hematochezia and edema. He had no prior history of renal or autoimmune disease. Symptoms developed one week after first dose of Moderna mRNA-1273 vaccine. Serum albumin was 1.6 g/dL and urine protein-creatinine ratio 8.5. Endoscopy showed esophagitis and small bowel ulcerations. Renal biopsy showed focally crescentic and necrotizing proliferative glomerulitis with IgA dominant deposits consistent with IgA vasculitis with renal involvement. He received pulse dose solumedrol followed by prednisone taper. Cyclophosphamide was initiated but then stopped due to cytopenias. After several weeks GI bleeding resolved spontaneously and proteinuria improved but remained in the nephrotic range. The course was further complicated by positive COVID testing prior to discharge-he was asymptomatic and received sotrovimab. On followup three months later the patient's edema resolved and serum albumin normalized. He continues to have subnephrotic range proteinuria with 2.3 grams/24 hours and active urinary sediment with dysmorphic red cells. He remains on steroid taper with plan for repeat renal biopsy to inform decisions regarding further immune suppression. He has not been rechallenged with COVID vaccine. Discussion(s): Rare associations between COVID vaccination and both de-novo and relapsing glomerular lesions, including minimal change disease, IgA nephropathy, ANCA and anti GBM glomerulonephritis, have been reported. COVID infection itself has been associated with an FSGS type lesion termed COVID-associated nephropathy (COVAN). As highlighted by this case, unvaccinated or partially vaccinated patients are at increased risk for COVID infection. Thus even in those with known glomerular disease, vaccination should still be recommended. Studies are needed to determine if patients with off-target glomerulopathies can be safely rechallenged with COVID vaccine, whether the course of COVID vaccine-associated disease glomerular lesions mimic the primary glomerular disease and how to optimally manage these patients.

COVID-19 in Patients with Hematologic Diseases.

The COVID-19 outbreak had a strong impact on people's lives all over the world. Patients with hematologic diseases have been heavily affected by the pandemic, because their immune system may be compromised due to anti-cancer or immunosuppressive therapies and because diagnosis and treatment of their baseline conditions were delayed during lockdowns. Hematologic malignancies emerged very soon as risk factors for severe COVID-19 infection, increasing the mortality rate. SARS-CoV2 can also induce or exacerbate immune-mediated cytopenias, such as autoimmune hemolytic anemias, complement-mediated anemias, and immune thrombocytopenia. Active immunization with vaccines has been shown to be the best prophylaxis of severe COVID-19 in hematologic patients. However, the immune response to vaccines may be significantly impaired, especially in those receiving anti-CD20 monoclonal antibodies or immunosuppressive agents. Recently, antiviral drugs and monoclonal antibodies have become available for pre-exposure and post-exposure prevention of severe COVID-19. As adverse events after vaccines are extremely rare, the cost-benefit ratio is largely in favor of vaccination, even in patients who might be non-responders; in the hematological setting, all patients should be considered at high risk of developing complications due to SARS-CoV2 infection and should be offered all the therapies aimed to prevent them.

Efficacy and Safety of a Delayed Graft Function Protocol in Kidney Transplant Recipients

Purpose: To evaluate the efficacy and safety of a protocol increasing the net state of immunosuppression for adult kidney transplant recipients (KTR) with delayed graft function (DGF). Method(s): Single-center retrospective cohort of adult KTR with DGF transplanted from January 2017 to March 2021. Pre- vs post-DGF protocol implementation outcomes were evaluated. Protocol included cumulative 6 mg/kg rabbit antithymocyte globulin (rATG) induction, non-weight-based mycophenolate mofetil dosing (1000 mg bid), and higher goal tacrolimus trough (9-12 ng/mL). Pre-protocol patients received cumulative 4.5 mg/kg rATG. Efficacy outcomes were biopsy proven acute rejection (BPAR) and graft loss at 6 months. Safety outcomes were incidence of cytopenia, infection, and all-cause readmission at 6 months. Result(s): Eighty-nine DGF patients met inclusion criteria. Baseline characteristics were similar between groups, with median age (57+/-19) years and majority Hispanic (42.7%) males (61.8%) with a negative crossmatch (100%). Most post-protocol patients received 6 mg/kg cumulative rATG induction (71.4%) and mycophenolate mofetil 1,000 mg bid (80.3%) with therapeutic tacrolimus troughs by discharge (64.3%). Significantly less BPAR was observed post-protocol (7/56, 12.5% vs 10/33, 30.3%;p = 0.04). Of those with BPAR, significantly less post-protocol patients experienced T-cell mediated rejection (TCMR) than pre-protocol (2/7, 28.6% vs 9/10, 90.0%;p = 0.03). However, antibody-mediated (4/7, 57.1% vs 1/10, 10%) and mixed (1/7, 14.3% vs 0%) rejection were more frequent post-protocol (p = 0.10 and 0.41, respectively). Graft loss was similar post- vs pre-protocol (5/56, 8.9% vs 0;p = 0.16). All post-protocol graft losses were due to death (4 from COVID-19 and 1 unknown). Safety outcomes were similar between groups (Table 1). Conclusion(s): An increased net state of immunosuppression in DGF KTR significantly lowered the 6-month incidence of BPAR without significantly affecting safety. TCMR incidence was significantly decreased, but displaced by antibody-mediated and mixed rejection, implying need to conduct further prospective studies of larger sample sizes. Given majority of graft losses were due to COVID-19 pneumonia, studies are needed to evaluate the risk of COVID-19 infections in DGF KTR, especially with the availability of vaccines. (Table Presented).