T-cell Imbalance or Decreased Th:Tc Ratio in Immune Thrombocytopenia: Is it Clinically Significant?

Background: Immune thrombocytopenic purpura (ITP) is an autoimmune disorder characterized by low platelet count and various contributing factor. The imbalance in T cells may also cause ITP. Therefore, the present study was planned to assess the role of  T-cells in pathogenesis of ITP and also to evaluate any possible link of Th:Tc imbalance to disease severity.Methods: The present study was conducted  with 111 patients of ITP and similar number of controls as case control study with 1:1 ratio of from January 2017 to July 2019. The patients were grouped according to the guideline of ASH as  newly diagnosed/persistent ITP(ND-ITP/P-ITP) and chronic/refractory ITP(C-ITP/R-ITP).The blood samples were obtained, and CBC parameters were observed using advanced hematology analyzer XN-1000.The T cells subset analysis was evaluated by BD FACS Calibur flow cytometer. The Fisher exact test was done to evaluated the difference among the groups with (p=<0.05) by using SPSS version 19.Results: Significantly reduced levels of hemoglobin, platelet counts with elevated IPF were observed in ND-ITP/P-ITP and C-ITP/R-ITP patients (p=<0.001).The significantly low Th:Tc ratio (p=<0.001) predicts imbalance of T cells in ND-ITP/P-ITP (0.86±0.47) as compared to control group (1.73±0.46).The mean of 0.84±0.34 Th:Tc ratio was observed in C-ITP/R-ITP children with ≤16 years. An insignificant difference (p= 0.89) was linked between children with non-severe chronic (0.84±0.42), severe chronic (0.82±0.49) and refractory ITP (0.85±0.51).Conclusion: In ITP patients’ low levels of Th:Tc ratio was observed suggesting dysregulation of immune system. The chronicity of the disease may be linked to elevated production of Tc in children (≤16 years) with C-ITP/R-ITP.Keywords: Immune thrombocytopenia; Platelets counts; Helper T-cells (Th); Cytotoxic T-cells (Tc) 

Bleeding disorders in the tribe: result of consanguineous in breeding

<p>Abstract</p> <p>Objective</p> <p>To determine the frequency and clinical features of bleeding disorders in the tribe as a result of consanguineous marriages.</p> <p>Design</p> <p>Cross Sectional Study</p> <p>Introduction</p> <p>Countries in which consanguinity is a normal practice, these rare autosomal recessive disorders run in close families and tribes. Here we describe a family, living in village Ali Murad Chandio, District Badin, labeled as haemophilia.</p> <p>Patients & Methods</p> <p>Our team visited the village & developed the pedigree of the whole extended family, up to seven generations. Performa was filled by incorporating patients, family history of bleeding, signs & symptoms, and bleeding from any site. From them 144 individuals were screened with CBC, bleeding time, platelet aggregation studies & RiCoF. While for PT, APTT, VWF assay and Factor VIII assay, samples were kept frozen at -70 degrees C until tested.</p> <p>Results</p> <p>The family tree of the seven generations comprises of 533 individuals, 63 subjects died over a period of 20 years and 470 were alive. Out of all those 144 subjects were selected on the basis of the bleeding history. Among them 98(68.1%) were diagnosed to have a bleeding disorder; 44.9% patients were male and 55.1% patients were female. Median age of all the patients was 20.81, range (4 months- 80 yrs). The results of bleeding have shown that majority had gum bleeding, epistaxis and menorrhagia. Most common bleeding disorder was Von Willebrand disease and Platelet functional disorders.</p> <p>Conclusion</p> <p>Consanguineous marriages keep all the beneficial and adversely affecting recessive genes within the family; in homozygous states. These genes express themselves and result in life threatening diseases. Awareness, education & genetic counseling will be needed to prevent the spread of such common occurrence of these bleeding disorders in the community.</p

Molecular analysis of eight severe FV‐deficient patients in Pakistan: A large series of homozygous for frameshift mutations

International audienc

Genotyping of five Pakistani patients with severe inherited factor X deficiency

International audience: Congenital factor X deficiency is a rare coagulation defect characterized by variable bleeding tendency. The aim of the study was to give a first insight of F10 gene mutations in Pakistani probands. Direct sequencing and/or next-generation sequencing was performed on the coding regions, boundaries and 5' and 3' untranslated regions of the F10 gene in five severe factor X-deficient patients from Pakistan. All patients were born from consanguineous marriages and displayed FX:C levels below 2%. Sequencing revealed five different substitutions, including three previously reported p.Ala15Asp, p.Gly406Ser, and p.Gly420Arg missense variants, and also two novel variants: p.Cys57Arg and p.Gln175*. Though one genotype could not be characterized, we were able to confirm the inherited nature of the defect using familial studies. As the copy number variations were ruled out, we hypothesized the presence of deep intronic mutants that might have escaped detection from sequencing or abnormalities in epigenetic regulation. Three patients presented with severe clinical symptoms, in the early days of life, whereas two presented only with trauma-provoked bleeds and bruises later in life. Those patients with milder forms bore the p.Gly406Ser at the homozygous state and F10 unknown alleles, respectively. F10 mutation spectrum in Pakistan is heterogeneous as seen in other populations. Identification of the F10 mutations is important for genetic counseling and prenatal diagnosis in subsequent pregnancies

Type and frequency of hemoglobinopathies, diagnosed in the area of Karachi, in Pakistan

Hemoglobinopathies are one of the major problems in Pakistan. A retrospective analysis of blood samples of 2731 patients from 2010 to 2014 was done at National Institute of Blood Disease & Bone Marrow Transplantation for the workup of anemia or other blood-related disorders. Whole blood samples in EDTA were collected; complete blood counts with peripheral smears were prepared. Hemoglobin (Hb) electrophoresis on Genio was performed at alkaline pH. Samples showing borderline results were further tested by high-performance liquid chromatography or for specific mutation analysis by ARMS-PCR. Out of total 2731, 935 (34.2%) patients had hemoglobinopathies. Out of these 935 patients who had hemoglobinopathies, beta thalassemia minor 51.8%, beta thalassemia major 24.1%, HbD trait 6.7, sickle/beta thalassemia 4.5%, sickle cell disease 3.9%, HbE trait 1.9%, and sickle cell trait 1.7% were most common hemoglobinopathies. Less prevalent were delta/beta thalassemia, HbE homozygous, HbD homozygous, and HbH disease

Validation of the ISTH/SSC bleeding assessment tool for inherited platelet disorders: A communication from the Platelet Physiology SSC

BACKGROUND: Careful assessment of bleeding history is the first step in the evaluation of patients with mild/moderate bleeding disorders, and the use of a bleeding assessment tool (BAT) is strongly encouraged. Although a few studies have assessed the utility of the ISTH-BAT in patients with inherited platelet function disorders (IPFD) none of them was sufficiently large to draw conclusions and/or included appropriate control groups. OBJECTIVES: The aim of the present study was to test the utility of the ISTH-BAT in a large cohort of patients with a well-defined diagnosis of inherited platelets disorder in comparison with two parallel cohorts, one of patients with type-1 von Willebrand disease (VWD-1) and one of healthy controls (HC). PATIENTS/METHODS: We enrolled 1098 subjects, 482 of whom had inherited platelet disorders (196 IPFD and 286 inherited platelet number disorders [IT]) from 17 countries. RESULTS: IPFD patients had significantly higher bleeding score (BS; median 9) than VWD-1 patients (median 5), a higher number of hemorrhagic symptoms (4 versus 3), and higher percentage of patients with clinically relevant symptoms (score &gt; 2). The ISTH-BAT showed excellent discrimination power between IPFD and HC (0.9 &lt; area under the curve [AUC] &lt; 1), moderate (0.7 &lt; AUC &lt; 0.9) between IPFD and VWD-1 and between IPFD and inherited thrombocytopenia (IT), while it was inaccurate (AUC 64 0.7) in discriminating IT from HC. CONCLUSIONS: The ISTH-BAT allows to efficiently discriminate IPFD from HC, while it has lower accuracy in distinguishing IPFD from VWD-1. Therefore, the ISTH-BAT appears useful for identifying subjects requiring laboratory evaluation for a suspected IPFD once VWD is preliminarily excluded