Common Hematological Abnormality Research Articles

Hematological Abnormalities in Culture Positive Neonatal Sepsis.

BackgroundIn neonatal sepsis, anemia, leukocytosis, thrombocytopenia, and a shortened coagulation time are the most common hematologic abnormalities. However, there is inadequate information regarding the hematological abnormalities in neonatal sepsis. Thus, we aimed to determine the magnitude of hematological abnormalities in neonatal sepsis.MethodsThis is a cross-sectional study that included 143 neonates with culture proven sepsis aged 1–28 days from September 2020 to November 2021 at the University of Gondar Specialized Referral Hospital. The sociodemographic data was collected using a pre-tested structured questionnaire, and the clinical and laboratory data was collected using a data collection sheet. A total of 2 mL of venous blood was taken using a vacutainer collection device for the complete blood count (CBC) and blood culture analysis. A univariate and multivariate logistic regression model was used to investigate factors associated with hematological abnormalities in neonatal sepsis. Statistical significance was declared when a p-value was less than 0.05.ResultsThe prevalence of anemia, thrombocytopenia, and leucopenia in neonatal sepsis was 49% (95% CI: 40.89–57.06), 44.7% (95% CI: 36.8–52.9), and 26.6% (95% CI: 22.01–29.40), respectively. On the other hand, leukocytosis and thrombocytosis were found in 7.7% (95% CI: 4.35–13.25) and 11.9% (95% CI: 7.56–18.21), respectively. Being female (AOR: 3.3; 95% CI: 1.20–3.82) and being aged less than 7 days (AOR: 2.44; 95% CI: 1.6–6.9) were found to be significant predictors of anemia.ConclusionThe magnitude of anemia, leucopenia, and thrombocytopenia is high in neonatal sepsis. Furthermore, being female and being younger than 7 days were risk factors for anemia. Thus, the diagnosis and treatment of anemia, leucopenia, and thrombocytopenia prevents further complications in neonatal sepsis.

Immuno-Haematological Abnormalities of HIV-Infected Patients Before and After Initiation of Highly Active Antiretroviral Therapy in the Antiretroviral Therapy Clinics of Six Health Facilities at Dessie Town, Northeast Ethiopia.

BackgroundIn people living with the human immunodeficiency virus, haematological abnormalities have been linked to an increased risk of disease progression and mortality. Hematological parameters may have a positive or negative impact on antiretroviral therapy. The aim of this study was to assess the immuno-haematological abnormalities of HIV-infected patients before and after the initiation of highly active antiretroviral therapy in the antiretroviral therapy clinics of six health facilities in Dessie, Northeast Ethiopia.MethodsA facility-based cross-sectional study was conducted from April to May 30, 2021, at the antiretroviral therapy clinics of six health facilities in Dessie Town. A total of 378 HIV-infected patients taking highly active antiretroviral treatment for at least 6 months by using a consecutive sampling technique were included. A well-organized questionnaire was used to collect socio-demographic and clinical information. Immune-haematological parameters were tested using a Mindray BS-300 hematology analyzer and a BD FACS count CD4 analyzer. Statistical analysis was performed using SPSS version 25 statistical software. Statistical significance was defined as a P-value of 0.05 with a 95% confidence interval.ResultsLeukopenia was found in 26.7% and 16.5%, neutropenia in 16.5% and 9.4%, lymphopenia in 20% and 3.1%, and thrombocytopenia in 25.9% and 7.1% of HIV patients before and after HAART initiation, respectively. There was a significant difference in total white blood cell, absolute neutrophil, red blood cell, hemoglobin value, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, red cell distribution width, platelet and CD4+ T cell counts in HIV patients before and after the initiation of HAART with P < 0.05.Conclusion and RecommendationAnemia, leukopenia, neutropenia, lymphopenia, and thrombocytopenia were the most common haematological abnormalities found in this study before and after HAART initiation. The prevalence of thrombocytopenia, immunosuppression, and viral load was reduced considerably after starting HAART.

Common Hematological Abnormalities in patients with Malaria presenting at Saidu Teaching Hospital, Swat Pakistan

Objectives: To determine the frequency of common hematological abnormalities in patients with malaria. Study Design: A cross-sectional study. Place and Duration of the Study: Department of Medicine, Saidu Teaching Hospital, Swat Pakistan from November 2019 to May 2020. Material and Methods: A total of 137 patients of both genders aged between 18-60 years and who presented with malaria in emergency or outpatient department were analyzed. Ten cc blood was taken from all enrolled patients and sent to institutional laboratory for the detection of anemia, thrombocytopenia, leucopenia and disseminated intravascular coagulation (DIC). Frequency of common hematological abnormalities among patients of malaria was noted. For data analysis, SPSS version 26.0 was used. Results: In a total of 137 patients, 95 (69.3%) were male and 42 (30.7%) female. Overall, mean age was 29.2±13.18 while 84 (61.3%) patients were aged between 18 to 30 years. Mean duration of disease was 1.2±1.09 weeks while 89 (65.0%) patients had duration of disease &lt;1 week. Table-1 is showing baseline characteristics of all patients. Anemia was found in 121(88.3%) patients, thrombocytopenia in 67(48.9%), leucopenia in 20 (14.6%) and 35 (25.5%) had DIC. Conclusion: The frequency of hematological abnormalities in patients with malaria was high. Anemia was found in 88.3% malaria cases, thrombocytopenia 48.9%, leukopenia 14.6% and DIC 25.5%. Keywords: Anemia, Leukopenia, malaria, thrombocytopenia.

Higher hemoglobin levels are associated with better physical performance among older adults without anemia: a longitudinal analysis

BackgroundAnemia is the most common hematological abnormality among older adults, and it is associated with decreased physical performance. But the role of hemoglobin in the absence of anemia remains unclear. Thus, this study aimed to assess the impact of hemoglobin levels on physical performance in Brazilian older adults without anemia.MethodsThe study is longitudinal in that it relies on two waves of the Saúde, Bem-Estar e Envelhecimento (SABE; Health, Well-being, and Aging) study: 2010 and 2015-2016. Mixed-effects linear regression was used to determine the effects of the hemoglobin concentrations on the Short Physical Performance Battery-SPPB over time among the 1,023 who had complete data and did not have anemia in 2010. In the follow-up, there were 567 without anemia.ResultsIn analyses adjusted for age, education, comorbidities, body mass index, and physical inactivity, we found a differential association between hemoglobin concentration and SBBP by sex, with a positive interaction (β Hb*female= 0.20, 95% CI 0.04,0.37). At lower levels of hemoglobin, women have lower levels of SPPB than men, but at higher levels of hemoglobin concentration, there are no sex differences in physical performance. In addition, higher age was negatively associated with SPPB levels and cardiometabolic diseases, other diseases, and physical inactivity. Education was positively associated with physical performance.ConclusionOur study demonstrates that higher hemoglobin levels were associated with better physical performance among older adults without anemia in Brazil. However, there were sex differences in this association. This finding is important because, in clinical practice, most health professionals focus on the World Health Organization definition of anemia. Our study suggests the importance of hemoglobin levels among older adults, even those without anemia, and highlights sex differences.

The role of platelet parameters for the diagnosis of preeclampsia among pregnant women attending at the University of Gondar Comprehensive Specialized Hospital antenatal care unit, Gondar, Ethiopia.

BackgroundPreeclampsia (PE) is a pregnancy‐related illness characterized by high blood pressure (BP) and proteinuria after the 20th gestational week (GW). Platelet (PLT) parameter changes are the common hematological abnormalities observed in PE patients. The main aim of this study was to assess the role of PLT parameters for PE diagnosis among pregnant women.MethodsA comparative cross‐sectional study was conducted at the University of Gondar Specialized Hospital. A total of 126 pregnant women (63 normotensive [NT] and 63 PE) were recruited using a convenient sampling method. Three milliliter blood was collected from each participant, and PLT parameters were determined using Sysmex XS‐500i analyzer. An independent t‐test supplemented with receiver‐operating characteristics (ROC) were used for comparisons and diagnostic value of PLT parameters between the study groups.ResultsPlatelet count (PC) was significantly lower in the PE group compared to that in the NT group, whereas mean platelet volume (MPV), platelet large cell ratio (P‐LCR), and platelet distribution width (PDW) were significantly higher in PE. MPV had the largest area under the curve (AUC) [0.91: 95% CI; 0.85–0.96] followed by PC [0.79: 95% CI; 0.72–0.87]. MPV can differentiate PE patients from NT pregnant women at cut‐off value ≥12.10 fl (84.1% sensitivity and 87.3% specificity) while PC can indicate PE at a cut‐off value ≤176.5 × 109/L (65.1% sensitivity and 87.3% specificity).ConclusionA decreased PC and an increased MPV, P‐LCR, and PDW can be used as a simple, cost‐effective, quick, and reliable method of PE screening. Of them, MPV is the best indicator of PE.

ASSESSMENT OF BASIC HEMATOLOGICAL CHANGES AND CLINICAL SEVERITY IN CHRONIC LIVER DISEASE WITH PERIPHERAL BLOOD NEUTROPHIL LYMPHOCYTE RATIO.

Introduction: A wide variety of hematological manifestations occur in chronic liver disease. Mild to moderate anemia and thrombocytopenia are the most common hematological abnormalities. Neutrophil-Lymphocyte Ratio, a novel inammatory biomarker can be used to predict mortality independent of Child-Turcotte-Pugh score in liver cirrhosis. Material And Methods: A cross sectional prospective study was conducted in 100 patients to assess hematological abnormalities and to correlate Neutrophil - Lymphocyte Ratio (NLR) with Child-Turcotte-Pugh (CTP) score for prediction of clinical outcome. NLR and CTP score were correlated using Chi square test and Spearman correlation test. Results: Mild anemia was common in females (61%) whereas moderate anemia was common in males (45%). Anemia was seen in 86% and thrombocytopenia in 72% of cases. Hypochromic microcytic anemia was the most common morphological type of anemia seen in chronic liver disease. NLR had a signicant positive correlation with CTP score in this study (r=0.288 and p=0.003). Conclusions: Mild anemia and moderate thrombocytopenia were the most common hematological abnormalities in chronic liver disease. Early diagnosis and correction of these abnormalities will reduce mortality. Neutrophil-Lymphocyte ratio can be used as an independent prognostic tool to predict clinical severity in chronic liver disease.

ROLE OF RBC INDICES IN CATEGORIZATION OF ANEMIA IN ANTENATAL WOMEN

BACKGROUND:Anemia is one of the most common hematological abnormality diagnosed during pregnancy. In India, anemia is the second most common cause of maternal death. This study was undertaken to determine the prevalence of anemia in antenatal women and to analyse the risk factors for anemia in pregnancy. MATERIAL AND METHODS: This study was conducted on 500 pregnant women coming to a tertiary teaching hospital in western Uttar Pradesh, for their rst antenatal visit, over a period of one year. Blood samples were taken in plain vacutainers for serum ferritin estimation and in EDTAvacutainers for red cell indices and peripheral smear examination. RESULTS: Prevalence of anemia was found to be 55.8%. Majority of the women were moderately anemic, normocytic normochromic anemia being the commonest type. Prevalence of anemia was high in second and fourth or more gravida and increased with increase in age of gestation. Serum ferritin levels were found to be decreased in all the trimesters. Prevalence of anemia was more in vegetarian women and in those who were not taking iron and folic acid supplements. CONCLUSION: The prevalence of antenatal anemia is high in India as compared to developed countries. Diet, intake of iron and folic acid, gravidity and age of gestation have signicant role in causation of anemia.

An Unusual Case of Pancytopenia: The Lessons Learnt.

Pancytopenia is a common hematological abnormality encountered in clinical practice. We here report a 36-year-old male who presented to emergency department with complaints of weakness of bilateral lower limbs, burning sensation in all four limbs with history of loose stools, and vomiting 5 days back. The complete blood count of patient showed pancytopenia with no circulating atypical cells. Bone marrow examination performed showed nonspecific but characteristic findings. After excluding the possibility of infective etiology, a possibility of heavy metal toxicity was suspected in multidisciplinary meeting. The urine and blood levels of arsenic done came out very high, and a diagnosis of arsenic poisoning was made. Patient had multisystemic involvement with features characteristic of arsenic poisoning. The present case was a diagnostic challenge in face of nonforthcoming history. This case beautifully highlighted the importance of multidepartmental approach in such cases to arrive at unerring diagnosis and the unique bone marrow findings, although nonspecific were sufficient enough to indicate the possibility of acute insult to the hematopoiesis.How to cite this article: Nair RR, Singh PK, Sharma J, Gambhir I, Khanna S, Jain AK, et al. An Unusual Case of Pancytopenia: The Lessons Learnt. Indian J Crit Care Med 2022;26(1):141–144.

COVID-19-A Trigger Factor for Severe Immune-Mediated Thrombocytopenia in Active Rheumatoid Arthritis.

Thrombocytopenia is defined as a platelet count below 150,000/mm3 for adults. There is still controversy about whether individuals with platelet counts of 100,000/mm3 to 150,000/mm3 should be classified as having genuine thrombocytopenia or borderline thrombocytopenia. Thrombocytopenia is considered mild when the platelet count is between 70,000 and 150,000/mm3 and severe if the count is less than 20,000/mm3. Thrombocytopenia in rheumatoid arthritis is a rare complication, with an incidence estimated between 3 and 10%. The main etiological aspects include drug-induced thrombocytopenia and immune thrombocytopenic purpura. The most common hematological abnormalities in SARS-CoV-2 infection are lymphopenia and thrombocytopenia. It has been observed that the severity of thrombocytopenia correlates with the severity of the infection, being a poor prognosis indicator and a risk factor for mortality. COVID-19 can stimulate the immune system to destroy platelets by increasing the production of autoantibodies and immune complexes. Autoimmunity induced by viral infections can be related to molecular mimicry, cryptic antigen expression and also spreading of the epitope. During the COVID-19 pandemic, it is of great importance to include the SARS-CoV-2 infection in differential diagnoses, due to the increased variability in forms of presentation of this pathology. In this review, our aim is to present one of the most recently discovered causes of thrombocytopenia, which is the SARS-CoV-2 infection and the therapeutic challenges it poses in association with an autoimmune disease such as rheumatoid arthritis.

Circulating Megakaryocytes in Peripheral Blood: A Clue to Satisfactory Platelet Recovery Post Induction Chemotherapy in Acute Myeloid Leukemia

Thrombocytopenia is a common hematological abnormality in newly diagnosed acute myeloid leukemia (AML) which may be linked to the degree of marrow infiltration by leukemic blasts as well as altered cytokine milieu in a subset of AMLs. We report the presence of circulating megakaryocytes in peripheral blood as a sign of satisfactory platelet recovery post induction chemotherapy in a pediatric subject with a favourable risk WHO category I AML with synchronous extramedullary myeloid cell tumor; and present a brief review of literature on the role of platelet recovery in AML prognosis.

Clinicoepidemiological findings of childhood brucellosis in a tertiary care center in Central Anatolia: with the emphasis of hematological findings.

Human brucellosis is one of the most widespread zoonotic diseases that are presented with predominantly hematological manifestations. We aimed to evaluate the hematological findings of childhood brucellosis and to determine the predictive clinical findings and laboratory tests that might be related to hematologic involvement. We retrospectively analyzed the medical records of children with brucellosis between 1 January 2005 and 31 December 2018. We compared predictive clinical and physical examination findings and laboratory tests in patients with and without hematological involvement. A total of 212 patients (127 boys (59.9%)) with a mean age of 9.4±4.7 years were evaluated in this study. Blood cultures were performed in 161 (75.9%) patients and Brucella spp were isolated in 70 (43.4%) of them. Ninety-two (43.4%) patients had hematological involvement at least in one series. Anemia was detected in 66 (31.7%) patients, leukopenia in 22 (10.6%) and thrombocytopenia in 10 (4.8%). Four patients (1.9%) had pancytopenia. Age distrubutions of the patients with and without hematological involvement were similar (p=0.6). In patients presented with fever, hepatomegaly and splenomegaly, hematologic involvement was significantly higher (p < 0.05). Hematological involvement was higher in patients who had elevated aspartate aminotransferase and alanine aminotransferase concentrations (p < 0.05). Hematological involvement was higher in patients with positive blood culture (p=0.005). Six patients (2.8%) were treated with intravenous immunoglobulin at 1000 mg/kg/day for two days in addition to anti-brucellosis treatment. Hematological involvement in brucellosis is a common finding regardless of age, especially in febrile, bacteremic patients and in patients who had hepatosplenomegaly and elevated liver enzymes. Anemia is the most common hematological abnormality.

Magnitude and Associated Factors of Thrombocytopenia among Pregnant Women Attending Antenatal Care Clinics at Dessie Comprehensive Specialized Hospital, Northeast Ethiopia.

BackgroundThrombocytopenia is a common hematological abnormality during gestation. Pregnant women with severe thrombocytopenia may be associated with a higher risk of excessive bleeding during or after delivery. Therefore, the main aim of this study was to assess the magnitude and associated factors of thrombocytopenia among pregnant women attending antenatal care services at Dessie comprehensive and specialized hospital, Northeast Ethiopia.MethodsAn institution-based cross-sectional study was conducted from February to March 2021. Using a systematic random sampling technique, a total of 294 pregnant women were enrolled in the study. Structured interviewer-administered questionnaires were used to collect socio-demographic and clinical data of study participants. Four ml of venous blood were collected from each pregnant woman and a complete blood count was determined using DIRUI BF 6500 automated hematology analyzer. Data were entered into Epidata version 4.6.0 and then exported into SPSS version 24.0. Multivariate logistic regression was used to assess the association between dependent and independent variables. P-value < 0.05 was considered to be statistically significant.ResultsA total of 294 pregnant women who visited antenatal care services at Dessie comprehensive specialized hospital were included. The mean (±SD) age of the study participants was 29.7 (±6.1) years. The prevalence of thrombocytopenia among pregnant women was 9.9% (95% CI: 6.5, 13.6). A mild type of thrombocytopenia is the major type and accounted for 72.4% whereas moderate thrombocytopenia and severe thrombocytopenia accounted for 17.2% and 10.4% respectively among pregnant women. Multivariate logistic regression showed that urban residents (AOR: 0.206,95% CI, 0.055-0.748), gestational ages within the first trimester (AOR: 0.183, 95% CI, 0.057-0.593) and gestational ages within the second trimester (AOR = 0.264, 95% CI, 0.092-0.752) were significantly associated and independent predictors of thrombocytopenia in pregnant women.ConclusionIn this study, the prevalence of thrombocytopenia was 9.9% and the mild type of thrombocytopenia (72.4%) was higher than the other type of thrombocytopenia among pregnant women. In multivariate logistic regression analysis, residence and gestational age (trimester) were significantly associated with thrombocytopenia. Therefore, the platelet count should be routinely determined during the antenatal care visit for proper diagnosis and to minimize bleeding during and or after childbirth.

HIV Myelopathy- A cross-sectional study of constellation of bone marrow findings in HIV/AIDS

Introduction and Aim: Haematological manifestations in HIV disease is common and can happen at any phase during the disease course. Anemia and thrombocytopenia are the most frequent hematologic abnormalities and are associated with high morbidity and mortality. The objective of current study was to observe and analyse various spectrum of bone marrow changes and haematological abnormalities in HIV/AIDS and to correlate findings with CD4 count. Material and Methods: A total of 44 patients over a period of 5 years were included. Clinical findings, hematological profile, bone-marrow aspirate, biopsy findings and CD4 count of these patients were documented. The association between absolute lymphocyte count (ALC) and CD4 count were further established. Results: The most common clinical indication for bone-marrow aspiration and trephine biopsy was pancytopenia (47.3%), pyrexia of unknown origin (15.1%), and unresolving hepatosplenomegaly (13.6%). Anemia (72.7%) was commonest haematological abnormality. Bone marrow aspirate was normocellular in majority of patients. Marrow findings were correlated with CD4 count and were found to be statistically significant. Tri-lineage dysplasia was observed in 9.1% of patients, and megakaryocytic dysplasia being the commonest(61.4%). Histiocytic aggregates (27.3%) were noted among which 6.8% showed acid fast bacilli in Ziehl-Neelsen stain. Fungal stains revealed histoplasmosis in 4.5% patients. Conclusion: There was a strong negative association between presence of anemia and dysplasia and CD4 count. When CD4 was &lt;200/µL and ALC&lt;1000/mm3, presence of anemia and dysplasia affecting various cell lines were commonly observed; therefore, can be used as indicators to assess the severity of the disease.

Molecular detection of Hepatozoon species infections in domestic cats living in Germany.

Three species of protozoal Hepatozoon species (H felis, H canis and H silvestris) are known to infect cats in Europe. The objective of this study was to determine the prevalence of Hepatozoon species in samples from cats living in Germany that were submitted to a veterinary laboratory. The study included cats tested for Hepatozoon species by PCR between 2007 and 2020 by the Laboklin laboratory. Travel history and haematological results were documented for cats with positive test results. From 2018 onwards, a partial 18S rRNA Hepatozoon gene fragment was sequenced from cats with positive PCR results. Sixty-four of 931 cats (7%) tested positive for Hepatozoon species. Sex and age did not have a statistically significant impact. Sequencing was carried out for 16 samples and revealed H felis in all cases. All cats with positive test results and a relevant travel history had been imported from the Mediterranean or south-eastern Europe. There were no autochthonous infections with Hepatozoon species. Leukocytosis, haemoconcentration and anaemia were the most common haematological abnormalities. Although infections with Hepatozoon species in cats are usually subclinical, it may be useful to screen cats imported from the Mediterranean and south-eastern Europe for these pathogens to prevent local transmission cycles. There was no evidence of autochthonous infections in Germany; however, further investigations regarding a possible transmission of Hepatozoon species from infected cats to blood-feeding arthropods in Germany may be of interest. To avoid potential spread of the pathogens, ectoparasite prophylaxis is advisable.

Clinico-Pathological Profile of Cutaneous Lupus Erythematosus Patients: A Report from A Tertiary Care Center of Chattogram

Background : Nearly all epidemiologic studies have involved patients with Systemic Lupus Erythematosus (SLE). Few authors have investigated the characteristics of patients with Cutaneous Lupus Erythematosus (CLE). We aim to describe the epidemiologic, clinical, and immunologic characteristics of a series of patients diagnosed with CLE.&#x0D; Materials and methods : This is a descriptive retrospective cross-sectional study carried out using the register records of total 218 patients attending the ‘Lupus Clinic’ in Chittagong Medical College Hospital during the period 2010 and 2020. The disease activity and damage of CLE were assessed according to the Cutaneous Lupus Erythematosus Disease Area and Severity Index (CLASI).&#x0D; Results : There were 187 females (86%) and 31 males (14%) with the female: male ratio being 6.1:1. The mean age was 28 ± 10.06 (mean ± SD) ranging between 11 and 65 years. The Chronic Cutaneous Lupus Erythematosus (CCLE) patients accounted 154 (71%) followed by Acute Cutaneous Lupus Erythematosus (ACLE) 46 (21%) and Subacute Cutaneous Lupus Erythematosus (SCLE) 18 (8%). On the basis of CLASI score, 91 (42%) patients were in mild form, 85 (39%) in moderate form and 42 (19%) in severe state. In LE specific skin lesions, common manifestation was photosensitivity 198 (91%) followed by discoid rash 154 (71%) and maculo-papular lupus rash 55 (25%). Oral ulcer was seen in 49 (22%) patients and malar rash in 46 (21%) patients. Other observed LE specific skin manifestations were papulo squamous rash 11 (5%), Toxic epidermal necrolysis like lesions 7 (3%) and lichenoid lesions 6 (3%). Among LE nonspecific skin lesions, common manifestation was nonscarring alopecia 123 (56%) followed by Raynaud’s phenomenon 17 (8%) livedo reticularis 17 (8%) Vasculitis 15 (7%) Periungual telangiectasia 7 (3%) erythema multiforme 6 (3%) and leg Ulcer 5 (2%). Antinuclear Antibody (ANA) 132 (61%) was the most common autoantibody followed by anti-ds DNA 91 (42%) anti-Sm antibody 2 (1%) anti-phospholipid antibodies 9 (4%) and anti-RNP 3 (1%). Hematological manifestations were seen in 161 (73.85%) where erythrocyte sedimentation rate was the most common hematological abnormality 161(73.85%) followed by lymphopenia 126 (57.80%) leucopenia 113 (51.80%) thrombocytopenia 107 (49.10%) anemia 92 (42%) monocytopenia 37 (16.97%). Conclusions: CCLE was the most common subtypes of CLE. Patients with different subtypes of CLE have distinct clinical and pathological characteristics. The onset or concurrence of SLE mandates the involvement of other disciplines depending on organ involvement. In the absence of consensus on a definition that makes it possible to differentiate cutaneous forms of LE from SLE, the dermatologist’s role in the correct diagnosis and classification of such patients is fundamental.&#x0D; Chatt Maa Shi Hosp Med Coll J; Vol.20 (2); July 2021; Page 79-84

Overexpression of Human TLR8 Causes Fatal Anemia in SLE-Prone Mice By Altering the Bone Marrow Erythropoietic Niche

Anemia is a common hematologic abnormality in patients with active systemic lupus erythematosus (SLE), a disease characterized by innate immune system activation by nucleic acid containing immune complexes and cell debris. Work in mouse models has shown that overactivation of several cytoplasmic innate immune sensors by either self-DNA or self-RNA specifically leads to erythroid cell death in the fetal liver while sparing other hematopoietic cell lineages.The endosomal receptors TLR7 and TLR8 both recognize ssRNA in humans. TLR7 overexpression in mice causes a lupus syndrome that includes the development of mild to moderate anemia (Hb >10) and thrombocytopenia and is associated with spleen histiocytosis, autoimmune hemolysis, erythrophagocytosis and compensatory stress erythropoiesis in the spleen. A recent study demonstrated that patients with TLR8 gain of function mutations present with immunodeficiency, inflammation and bone marrow failure. However the role of TLR8 in SLE has been difficult to study in mice because it has a 5 amino acid deletion that attenuates its RNA binding capacity.To address the role of TLR8 in SLE, we overexpressed human TLR8 in a lupus mouse model (huTLR8tg.Sle1.Yaa) using a BAC transgene. 50% of homozygous huTLR8tg.Sle1.Yaa mice developed severe anemia (Hb<9) resulting in early mortality starting at 3-4.5 months of age. This phenotype required both the Sle1 and Yaa loci that promote the formation of high titer anti-chromatin and anti-RNA antibodies and onset of nephritis at >6 months of age. There was no difference in autoantibody titers between Sle1.Yaa wt and huTLR8tg mice and early death was not due to premature onset of renal disease. All mice had normal RBC indices prior to 10 weeks of age. Anemia was associated with an increase in bone marrow (BM) trabecular bone and a decrease in erythroblastic islands (EBI) in the BM with compensatory stress erythropoiesis leading to reticulocytosis and vast splenomegaly. RBC half-life was normal even after the development of reticulocytosis, but decreased in severely anemic mice as a pre-terminal event. Using CFSE labeling of RBCs we showed that hemophagocytosis occurred in vivo as a result of expansion of phagocytic red pulp macrophages.Flow cytometry of BMs from transgenic mice showed normal erythroid progenitors but a block at the late CFU-E/early proerythroblast stage leading to overall decreased terminal erythroid differentiation. Single cell RNAseq of EBIs isolated from transgenic BMs confirmed the block in erythropoiesis. The erythroblast cluster proximal to the block had a signature of mitochondrial stress and decreased proliferation. Spleen EBIs from transgenic mice were characterized by a low frequency of early erythroblasts compared with BM EBIs. We found 6 related clusters of EBI central macrophages in the BMs of both wt and transgenic mice. In transgenic mice, most of these displayed an inflammatory and Type 1 IFN signature. One cluster (M2) expressed all the classical central macrophage phenotypic markers in wt mice but was characterized in transgenic mice by downregulation of multiple phagocytic receptors and a 5-fold decrease of VCAM1 expression. Loss of VCAM1 and downregulation of CD169 in central macrophages of transgenic BMs was confirmed by flow cytometry. By contrast spleen central macrophages from transgenic mice retained VCAM and CD169 expression.Together, these results suggest that failure of BM erythropoiesis in huTLR8 transgenic SLE.Yaa lupus-prone mice, in which the acquisition of anti-nucleic acid autoantibodies drives excess innate stimulus through TLR7 and huTLR8, is due to a block in differentiation from CFU-E to the early proerythroblast stage; this is associated with an inflammatory phenotype specifically in BM erythroblastic island central macrophages and down regulation of adhesion and phagocytic receptors. Stress erythropoiesis in the spleens is associated with vast expansion of red pulp macrophages with phagocytic properties and fatal anemia is associated with a decrease in red blood cell half-life, suggesting that excessive RBC phagocytosis, coupled with insufficient erythroblast progenitors, eventually exceeds the capability of stress erythropoiesis to replace the RBC mass. DisclosuresKalfa: Agios Pharmaceuticals, Inc.: Other: Steering Committee, Research Funding; FORMA Therapeutics, Inc: Research Funding. Paulson: Forma Therapeutics: Consultancy. Blanc: Keros Therapeutics, Inc.: Consultancy.

Tandem CD19/CD22 Dual Targets CAR T-Cells Bridging Hematopoietic Stem Cells Transplantation Acquires Robust Remission for Relapsed and Refractory B Acute Lymphoblastic Leukemia Patients

Background: CD19 chimeric antigen receptor T (CAR-T) cells have demonstrated impressive early response rates in relapse and refractory B acute lymphoblastic leukemia (r/r B-ALL). However, a high rate of patients suffered a relapse, which occurred in a large subset of other trials and confers dismal outcomes. Dual targets approaches are proved to optimize the response rate and prevent CD19 - relapse. Alternatively, limited patients accepted the tandem CD19/CD22 CAR-T therapy, but the clinical trials in a large scale to investigate the tandem CAR-T were rare.Methods: We conducted an open label, single center clinical trial at the First Affiliated Hospital of Soochow University to investigate the efficacy and safety of tandem CD19/CD22 dual targets CAR T-cells for r/r B-ALL. All patients received FC (fludarabine, 30 mg/m2, days 1-3 and cyclophosphamide, 300 mg/m2, days 1-3) based chemotherapy as the lymphodepleting regimen. Median infusion dose of CAR-T cells was 1(0.5-2.5) *10 7 cells/kg.Results: From October 2017 to June 2021, a total of 47 patients were treated with CD19/CD22 CAR T-cells and included in our analysis.Among the 47 patients, primary refractory B-ALL patients account for 44.68%. 27 patients (57.4%) had a high disease burden, with 20% or more blasts in BM. Consequently, at day 28 assessment, 47 patients (100%) got hematological CR and the 40 out of 47 patients (85.1%) achieved minimal residual disease complete remission (MRD -CR).The toxicities of CD19/CD22 CAR T-cells therapy were reversible and clinically manageable. Cytokine reverse syndrome of any grades occurred in 41 of 47 patients (87.23%) and was severe (grade>2) in 8 (17.02%). Immune effector cell-associated neurotoxicity happened in one patient. The most common severe hematological abnormalities were grade 3/4 leukopenia (74.47%). Serious thrombocytopenia and anemia occurred in 48.93% and 57.44% patients. The non-hematological toxicity were reversible with tight monitoring and support care.Within a median follow-up of 21.83 months (range, 2.57 to 42.53), the median overall survival (OS) and leukemia free survival (LFS) for the entire cohort have not reached. The OS rate of all the patients was 93.569% (95%CI, 80.97% to 97.832%) at 6 months, 78.721% (95%CI, 60.719% to 87.625%) at 1 year and 74.578% (95%CI, 55.263% to 84.969%) at 2 years in all patients. The LFS rate was 87.031% (95%CI, 73.375% to 93.958%) and 68.297% (95%CI, 51.419% to 80.365%) at 6 month and 1 year, respectively. The 6-months cumulative incidence of relapse (CIR) was 8.96%, while 1-year CIR was 23.254%.Thirty-four of 47 patients (72.34%) proceeded to a bridging allogeneic hematopoietic stem cell transplantation (allo-HSCT). The OS of HSCT group was 94.118% at 6 months and 80.420% at 1 year. The 6-months OS of no-HSCT group was 83.916%, while 1-year OS was 74.592%. The HSCT group had significantly better LFS and lower CIR than the no-HSCT group (LFS, p=0.0459; CIR, p=0.0267). We initially performed multivariable Cox regression analyses, which shows that better long-term survival in patients with MRD -CR status, as well as bridging allo-HSCT.Conclusions: Tandem CD19/CD22 CAR-T cells are safety and highly effective in inducing CR for r/r B-ALL patients. The consolidative allo-HSCT can provide long-term durable disease control in these patients. DisclosuresNo relevant conflicts of interest to declare.

Development and optimization of paclitaxel loaded Eudragit/PLGA nanoparticles by simplex lattice mixture design: Exploration of improved hemocompatibility and in vivo kinetics

Anemia is the most common hematological abnormality of chemotherapy, which is responsible for poor clinical outcomes. To overcome this complication, the present study was aimed for developing a Eudragit/polylactic-co-glycolic acid (PLGA) based nanoparticulate system for a model drug paclitaxel (PTX). The study was planned using a simplex lattice mixture design. PTX nanoparticles (PTXNp) were evaluated in vitro for physicochemical properties, hemolytic effects and cytotoxic effects. Further, the nanoparticles were subjected to in vivo screening using rats for hemocompatibility, pharmacokinetic profile, and biodistribution to the vital organs. The PTXNps were 65.77–214.73 nm in size, showed more than 60% sustained drug release in 360 h and caused less than 8% hemolysis. The parameters like red blood cell count, activated partial thromboplastin time (aPTT), prothrombin time (PT) and C3 complement were similar to the negative control. Cytotoxicity results suggested that all the PTXNp demonstrated drug concentration-dependent cytotoxicity. The in vivo pharmacokinetic study concluded that PTXNp formulations had significantly higher blood AUC (93.194.55–163,071.15 h*ng/mL), longer half-lives (5.80–6.35 h) and extended mean residence times (6.05–8.54 h) in comparison to PTX solution (p < 0.05). Overall, the study provides a nanoparticulate drug delivery system to deliver PTX safely and effectively along with reducing the associated hematological adverse effects.

Magnitude and associated factors of anemia among AZT based HAART experienced adult HIV patients at University of Gondar Comprehensive Specialized Referral Hospital, Northwest, Ethiopia, 2019: a retrospective cohort study

BackgroundAnemia is the most common hematologic abnormalities in AIDS patients usually associated with disease progression and poor clinical outcomes. Zidovudine (AZT), which is one of the nucleoside reverse transcriptase inhibitor drug families of the first line antiretroviral therapy regimen for HIV/AIDS patients, causes anemia due to early long-term of higher-dose therapy. This study was aimed to assess the magnitude and associated factors of anemia among AZT containing HAART experienced adult HIV/ADIS patients at University of Gondar Comprehensive Specialized Referral Hospital, northwest, Ethiopia, 2019.MethodsA retrospective cohort study was conducted among a total of 320 adult AZT based HAART experienced HIV/AIDS patients from January 2016 to December 2018. Systematic random sampling technique was used to select the patients’ charts. All required data for this study were extracted from patients’ medical charts. Data were coded, cleared and entered into Epi Info version 3.5.3, and transformed to SPSS version 20 for analysis. Descriptive statistics, bivariable and multivariable logistic regression models were fitted to identify associated factors of anemia and P-value < 0.05 was considered as statistically significance.ResultsA total of 320 adult AZT based HAART experienced HIV/AIDS patients’ charts were assessed. Of the total patients, 198 (61.9%) were females and 133 (41.6%) were within the age range of 35–45 years. More than half, 237(76.9%) of the patients were from the urban area and 186 (58.1%) were on WHO clinical stage III at the baseline. The prevalence of anemia was 50% (95% CI 44.7–55.0%), 44.1% (95% CI 38.4–50.0%), 35.6% (95% CI 30.3–40.6%), 40% (95% CI 34.4–45.6%), 40.6% (95% CI 35.0–46.3) and 39.1% (95% CI 33.4–44.1%) at baseline, 6 months, 12 months, 18 months, 24 months and 30 months of follow-up period, respectively. The overall prevalence of anemia was 41.6%. Anemia had significant association with WHO clinical stage and base line Hgb values.ConclusionsA significant number of participants were anemic in this study. WHO clinical stage and baseline Hgb value were the contributing factors for anemia among these patients. Therefore, anemia needs an immediate intervention on associated factor to improve the anemic status and living condition of HIV patient.

Clinical profile of young females with systemic lupus erythematosus: an observational study

Background: Systemic lupus erythematosus (SLE) is an autoimmune disorder predominantly affecting women of child bearing age group and is known to require significant lifestyle modifications. The manifestations of SLE are myriad and it may virtually affect every system of affected individual. We Undertook this study to know the clinical profile of young female patients having SLE. The aim of the study was to study clinical features and medical therapy of young female patients having SLE.Methods: This was a prospective study in which 60 female patients diagnosed to be having lupus were included on the basis of a predefined inclusion and exclusion criteria. Investigations relevant to the diagnosis such as Antinuclear antibodies, Antiphospholipid Antibodies, Anti B2GP1 IgG, Anti-dsDNA antibody, Anti Smith Antibody and complement (C3 and C4) levels were done in all the cases. Other investigations such as imaging studies were done in selected cases. Clinical features and medical management being taken by these patients were analyzed. SSPS 21.0 software was used for statistical purpose.Results: The mean duration of the disease in studied cases was found to be 6.96±4.51 years. Malar rash was the commonest type of rash seen in these patients and was present in 53 (88.33%). Arthralgia with or without arthritis was seen in 54 (90%) of the cases. Anemia was the most common hematological abnormality and was seen in 17 (28.33%) patients. Renal involvement in the form of proteinuria was seen in 25 (41.67%) cases. The most common form of pulmonary involvement was pleural effusion which was seen in 4 (6.67%) patients. Cardiovascular manifestations were seen in 11 patients (18.33%).Conclusions: SLE usually affect women of child bearing age group and have a myriad clinical presentation. A thorough knowledge of various clinical presentations and a high index of suspicion is necessary to diagnose SLE particularly in its early stages.