scholarly journals Hereditary Spherocytosis

Blood ◽  
1951 ◽  
Vol 6 (11) ◽  
pp. 1099-1113 ◽  
Author(s):  
LAWRENCE E. YOUNG ◽  
RICHARD F. PLATZER ◽  
DONALD M. ERVIN ◽  
MARY JANE IZZO
Keyword(s):  
Peripheral Blood ◽  
Fanconi Syndrome ◽  
Congenital Abnormalities ◽  
Hereditary Spherocytosis ◽  
Osmotic Fragility ◽  
Test Tube ◽  
Red Cells ◽  
Blood Samples ◽  
Selective Retention ◽  
Multiple Congenital Abnormalities

Abstract 1. Three patients with hereditary spherocytosis and 1 patient with the Fanconi syndrome (pancytopenia and multiple congenital abnormalities) were transfused prior to splenectomy with normal erythrocytes of types which could be differentiated serologically from those of the recipients. The proportions of donated and patient’s cells in peripheral blood and in blood washed from the minced spleens were determined by differential agglutination, and the osmotic and mechanical fragilities of the two types of cells in peripheral and splenic blood were measured by differential agglutination of the corpuscles remaining in each test tube after partial hemolysis had occurred. 2. In each case of hereditary spherocytosis the proportion of recipient’s cells was much higher in splenic than in peripheral blood, indicating selective retention of the thicker corpuscles within the spleen. Osmotic fragility of thie patient’s red cells was much greater in samples of splenic mince blood than in peripheral venous samples, while the fragility of the donated red corpuscles was normal or nearly normal in both splenic and peripheral blood. In the patient exhibiting the Fanconi syndrome, on the other hand, neither the patient’s red cells nor donated red cells were retained to any extent in the spleen and the fragility of neither type of cell was altered. 3. Spleens removed surgically from 3 patients with idiopathic thrombocytopenic purpura were perfused with mixtures of normal A or B cells and group O cells drawn from a splenectomized individual with hereditary spherocytosis. During perfusion the spheroidal cells were selectively removed from the mixtures and at the end of each experiment red cells of this type predominated in the blood samples washed from the minced spleens. A fourth excised spleen was perfused with a mixture of two types of normal cells, neither of which was retained to any extent by the spleen during perfusion. The perfusion experiments show that spleens from patients with nonhemolytic disease are also capable of selective trapping of spheroidal cells. 4. The experiments described indicate that the spleen acts as a filter and trap and as an "incubator" in accelerating destruction of red corpuscles in patients with hereditary spherocytosis.

scholarly journals Physical Properties of Red Cells as Related to Effects in Vivo. III. Effect of Thermal Treatment on Survival of Red Cells in the Dog. Role of the Spleen

Blood ◽  
1968 ◽  
Vol 32 (6) ◽  
pp. 872-883 ◽  
Author(s):  
DENNIS J. CARLSON ◽  
THOMAS HALE HAM
Keyword(s):  
Sickle Cell Disease ◽  
Thermal Treatment ◽  
Peripheral Blood ◽  
Hereditary Spherocytosis ◽  
Osmotic Fragility ◽  
Red Cells ◽  
Cell Disease ◽  
Homozygous Sickle Cell Disease

Abstract Dog red cells heated at 49 C. for 15 minutes with no change in osmotic fragility, showed a decreased rate of survival in vivo, and increased sequestration by the spleen, and an increase in the osmotic fragility when recovered from the spleen. The peripheral blood showed normal osmotic fragility at all times. These changes were comparable to those seen in the spleen in homozygous sickle-cell disease and hereditary spherocytosis. There was no hemoglobinemia. Splenectomy decreased the rate of destruction of such heated red cells in vivo. In these studies the rigidity of the red cells with increased viscosity, but with normal osmotic fragility, may have resulted in the removal of the heated red cells by the spleen and subsequent conditioning of these cells in the spleen. In contrast to the 15 minutes heating, dog cells heated at 49 C. for 60 minutes had increased osmotic fragility and were rapidly removed from the circulation. There was sequestration by the spleen or by the liver with hemoglobinemia. The red cells with the greatest increase in osmotic fragility were not preferentially removed in vivo.

Targeted Deletion of γ-Adducin in Mice.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 1723-1723
Author(s):  
Kenneth E. Sahr ◽  
Amy J. Lambert ◽  
Steven L. Ciciotte ◽  
Luanne L. Peters
Keyword(s):  
Western Blotting ◽  
Peripheral Blood ◽  
Osmotic Fragility ◽  
Membrane Skeleton ◽  
Red Cells ◽  
Null Mouse ◽  
Exon 2 ◽  
Rbc Membrane ◽  
Blood Smears ◽  
Peripheral Blood Smears

Abstract The adducins are a family of three closely related proteins (α, β, γ) encoded by distinct genes. α- and γ-adducin are expressed ubiquitously, while β expression is restricted to hematopoietic cells and the brain. In red blood cells (RBCs) adducin localizes to spectrin-actin junctions in the membrane skeleton as αβ heterotetramers. Previously (Gilligan et. al., PNAS, 1999) we showed that deletion of β-adducin results in osmotically fragile, microcytic RBCs and an overall phenotype of hereditary spherocytosis (HS). Notably, α-adducin was significantly reduced in β-adducin null RBCs. We also demonstrated that γ-adducin is present in low amounts in normal mouse RBCs and is upregulated ∼5-fold in β-adducin null RBCs. The increase in γ-adducin suggests that αγ heterotetramers may be compensating for the absence of β-adducin. In an effort to analyze γ-adducin function in RBCs in greater detail, we generated a conditional γ-adducin knockout allele in mice using a Cre-loxP strategy to delete exon 2 containing the start codon. All mice were maintained on a segregating B6.129 genetic background. Western blotting confirmed the absence of γ-adducin in spleen homogenates and RBC ghost preparations from γ-adducin null mice. All other membrane skeleton proteins examined by a combination of SDS-PAGE and western blotting, including α- and β-adducin, are normal in γ-adducin null RBCs (spectrin, ankyrin, band 3, protein 4.1, protein 4.2, dematin). Phenotypically, γ-adducin null mice display normal growth curves and show no overt defects. γ-adducin null RBCs appear normal on Wright’s stained peripheral blood smears and by scanning electron microscopy (SEM). The RBC count, hemoglobin content, hematocrit, MCV, reticulocyte %, osmotic fragility, and all other hematopoietic parameters are normal in γ-adducin null mice vs. wildtype. The apparent compensation by γ-adducin in β-adducin null red cells previously observed was tested by intercrossing mice null for γ- and β-adducin to produce βγ null double homozygotes. The additional loss of γ-adducin did not exacerbate the β-adducin null RBC phenotype as judged by examination of peripheral blood smears and SEM. Moreover, RBC osmotic fragility and complete blood counts in βγ-adducin null mice did not differ from β-adducin null mice. Western blotting of RBC ghost proteins confirmed reduction of α-adducin to ∼20% of normal in β-adducin null mice, as previously described. Strikingly, α-adducin in βγ-null RBC ghosts is reduced to barely detectable levels (<5% of normal). These studies show that (1) loss of γ-adducin alone does not significantly impact RBC membrane skeleton structure and function; (2) α- and β-adducin are stable and present at normal levels in the absence of γ-adducin; (3) the loss of γ-adducin in β-adducin null mice does not further exacerbate the β-adducin null HS phenotype; (4) the exacerbated loss of α-adducin in βγ double null RBCs suggests that up-regulated γ-adducin in β-adducin null mice associates in some way with and stabilizes α-adducin in the RBC membrane, but is unable to compensate functionally for the loss of the β subunit. We conclude that the normal function and stable incorporation of adducin into the peripheral membrane skeleton of red cells requires the presence of heterologous αβ binding subunits. Additional studies of adducin null mouse models, including our recently generated α-adducin null strain, will be useful tools in defining adducin functions and interactions in multiple tissues and organs.

scholarly journals Hereditary Spherocytosis

Blood ◽  
1951 ◽  
Vol 6 (11) ◽  
pp. 1073-1098 ◽  
Author(s):  
LAWRENCE E. YOUNG ◽  
MARY JANE IZZO ◽  
RICHARD F. PLATZER
Keyword(s):  
Body Temperature ◽  
Gene Mutation ◽  
Hereditary Spherocytosis ◽  
Osmotic Fragility ◽  
Red Cells ◽  
Laboratory Findings ◽  
Low Degree ◽  
Qualitative Test ◽  
Chronic Hemolytic Anemia

Abstract Clinical, hematologic and genetic data on 28 cases of hereditary spherocytosis are presented for the purpose of characterizing this disorder as completely as possible. On the basis of this experience it is recommended that the following typical laboratory findings be sought in establishing a diagnosis in suspected cases: (1) Presence of spherocytes or abnormally thick red cells in peripheral blood; (2) greater than normal osmotic fragility of the red cells; in cases in which the fragility of fresh cells is not significantly increased, determinations should be made after sterile incubation of the blood at body temperature for 24 hours; (3) greater than normal mechanical fragility of freshly drawn red cells; (4) negative antiglobulin (Coombs) test; (5) greater than normal lysis of the red cells during sterile incubation at body temperature for 48 hours; and (6) presence of similar abnormalities in relatives. Abnormality of the erythrocyte persisted in all of the 11 patients in this series followed one or more years after splenectomy. An unusual case of chronic hemolytic anemia is described but not included in the numbered series because (1) both parents were hematologically normal and (2) spherocytosis and abnormally great osmotic and mechanical fragility and autohemolysis could not be demonstrated after the fifth postoperative month. Classification of this case is deferred pending further experience. Demonstration in a parent, sibling or offspring of red cells showing the afore-mentioned abnormalities is necessary for an unequivocal diagnosis, but this requirement cannot always be met because relatives may not be available for examination. Moreover, when parents and/or several siblings are examined without positive findings, low gene expressivity, gene mutation and illegitimacy may be considered as explanations. Evidence is cited to suggest the possibility of a low degree of penetrance or expression in some cases and to illustrate the need for still more sensitive laboratory tests that might aid in diagnosis of the mildest forms of this disease. The lower incidence of spherocytosis in siblings of propositi than in offspring of propositi is cited as evidence bearing on the theory of gene mutation in some propositi. A simplified "qualitative" test of osmotic fragility of incubated red cells is described.

A Case of Congenital Red Cell Pyruvate Kinase Deficiency Associated with Hereditary Spherocytosis

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5272-5272
Author(s):  
Cristina Vercellati ◽  
Anna Paola Maria Luisa Marcello ◽  
Elisa Fermo ◽  
Paola Bianchi ◽  
Carla Boschetti ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Pyruvate Kinase ◽  
Hemolytic Anemia ◽  
Peripheral Blood ◽  
Blood Smear ◽  
Hereditary Spherocytosis ◽  
Osmotic Fragility ◽  
Peripheral Blood Smear ◽  
Red Cell ◽  
Red Cell Membrane

Abstract Abstract 5272 Pyruvate kinase (PK) deficiency, transmitted as an autosomal recessive trait, is the most common erythroenzymopathy of glycolytic pathway (prevalence of 1:20,000) associated with chronic non spherocytic hemolytic anemia from mild to severe. More than 180 mutations in the PK-LR gene have been so far reported, and genotype-phenotype correlation has been established for some of them. Hereditary Spherocytosis (HS) is the most common congenital hemolytic anemia in Caucasians, with an estimated prevalence ranging from 1:2000 to 1:5000. The main clinical features are hemolytic anemia from compensated to severe, variable jaundice, splenomegaly and cholelythiasis. The molecular defect is highly heterogeneous, caused by proteins involved in the attachment of cytoskeleton to the membrane integral domain (spectrin, ankyrin, band 3 and protein 4.2). We describe a case of PK deficiency associated with HS. The propositus was a 13 years-old Italian male with neonatal jaundice and need of blood transfusion (Hb 5.8 g/dL) during an infectious episode. At the time of the study Hb was 13.9 g/dL, MCV 81.8 fL, reticulocytes 207×109/L, unconjugated bilirubin 2.16 mg/dL, LDH 605 U/L, haptoglobin <20 mg/dL. The peripheral blood smear examination showed the presence of spherocytes (16%) and some ovalocytes (2%). The study of the most important red cell enzymes revealed reduced PK activity (59% of normal). Direct sequencing of PK-LR gene showed compound heterozygosity for the 994A mutation (Gly332Ser) and the −148T variant localized the erythroid specific promoter region. The presence of spherocytes in peripheral blood smear prompted us to investigate for the coexistence of HS. Erythrocyte osmotic fragility was decreased and SDS–PAGE analysis of red cell membrane proteins revealed a 30% spectrin reduction. Family study demonstrated a heterozygous condition for the 994A mutation in the father, who also displayed comparable enzyme deficiency, whereas promoter variant −148T was detected in the mother and in the brother. No red cell membrane abnormalities were present in the family members, although positive EMA binding test and increased osmotic fragility were found in the father and brother. The co-existence of HS and PK deficiency is very rare event, only few cases are described to date. Clinical, family and molecular studies allowed the determination of the interrelationship between the two RBC abnormalities in the patient and his relatives. The reduced PK activity in the propositus and his father is justified by heterozygous 994A mutation. The more severe clinical picture in the propositus could be caused by the coexistence of HS and by the presence of −148T mutation, that although it seems not to have effects on PK-LR mRNA expression, is often detected in PK deficient subjects with heterozygous PK mutations. Disclosures: No relevant conflicts of interest to declare.

Evaluation of a Flow-Cytometric Osmotic Fragility Test for Hereditary Spherocytosis in Chinese Patients

2015 ◽  
Vol 135 (2) ◽  
pp. 88-93 ◽  
Author(s):  
Yi-Feng Tao ◽  
Zeng-Fu Deng ◽  
Lin Liao ◽  
Yu-Ling Qiu ◽  
Xue-Lian Deng ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cells ◽  
Cell Morphology ◽  
Red Blood Cell ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Hereditary Spherocytosis ◽  
Osmotic Fragility ◽  
Flow Cytometric ◽  
Blood Cell Morphology

Background: Osmotic fragility testing based on flow cytometry was recently introduced for the screening of hereditary spherocytosis (HS). This study was undertaken to evaluate the clinical diagnostic value of a flow-cytometric osmotic fragility test for HS. Methods: Peripheral blood was collected from 237 subjects at the First Affiliated Hospital of Guangxi Medical University, including 56 HS patients, 86 thalassemia patients and 95 healthy controls. The samples were examined by flow-cytometric osmotic fragility test and the percentage of residual red blood cells was used to determine HS. Peripheral blood smears were performed to examine the red blood cell morphology. Results: With clinical diagnosis of HS as the gold standard and the percentage of residual red blood cells <23.6% as the diagnostic threshold in the flow-cytometric osmotic fragility test, the sensitivity of the flow-cytometric osmotic fragility test for HS was 85.71% and the specificity was 97.24%. Conclusion: The flow-cytometric osmotic fragility test combined with a red blood cell morphology test by peripheral blood smear could be a simple, practical and accurate laboratory screening method for HS.

scholarly journals Studies on Spontaneous In Vitro Autohemolysis in Hemolytic Disorders

Blood ◽  
1956 ◽  
Vol 11 (11) ◽  
pp. 977-997 ◽  
Author(s):  
LAWRENCE E. YOUNG ◽  
MARY JANE IZZO ◽  
KURT I. ALTMAN ◽  
SCOTT N. SWISHER
Keyword(s):  
Lactic Acid ◽  
Hereditary Spherocytosis ◽  
Substantial Reduction ◽  
Osmotic Fragility ◽  
Red Cells ◽  
Hemolytic Disease ◽  
Myeloid Metaplasia ◽  
Effect Of Glucose

Abstract Measurements of spontaneous lysis (autohemolysis) of red cells in sterile defibrinated blood after 48 or more hours of incubation at body temperature were found useful in the investigation of certain hemolytic states. Abnormally rapid autohemolysis was demonstrated most consistently in hereditary spherocytosis, but was also found in other types of spherocytosis and in several examples of non-spherocytic hemolytic anemia. Autohemolysis above the normal range can be regarded as strong presumptive evidence of a hemolytic disorder, but a normal rate of autohemolysis by no means excludes the possibility of a hemolytic process in any given case. Abnormalities of hereditary spherocytes causing increased autohemolysis were shown to be correlated with those responsible for spherocytosis as reflected in osmotic fragility tests. There appeared to be closer correlation with abnormalities causing increased osmotic fragility of the cells after 24 hours incubation. The autohemolysis test and the osmotic fragility test on incubated red cells were found to be equally sensitive in their capacity to detect spherocytosis of the hereditary type. Addition of adenosine, guanosine or inosine caused moderate to marked reduction in autohemolysis of nearly all types of red cells tested. Lysis of incubated spherocytes from active cases of autoimmune hemolytic disease was more markedly inhibited by a small amount of adenosine than was the lysis of hereditary spherocytes. Glucose regularly caused marked inhibition of autohemolysis of hereditary spherocytes and of red cells from patients with acute leukemia. The effect of glucose on HS red cells might have been due in part to a lowering of pH by formation of lactic acid since acidification of the blood with lactic, citric or hydrochloric acid also caused substantial reduction in autohemolysis. Glucose caused slight to marked increase in lysis of red cells in certain other cases of spherocytosis, notably in autoimmune hemolytic disease and in myeloid metaplasia. Addition of lactic acid to the blood from several of these patients had similar effect. In 2 atypical cases of chronic spherocytosis, with hematologically normal relatives, autohemolysis was accelerated by addition of glucose to the blood samples. Adenosine had little effect until after splenectomy when both glucose and adenosine inhibited hemolysis, much the same as in blood from typical cases of hereditary spherocytosis. It seems likely that when the abnormalities of carbohydrate metabolism of the red cells in certain hemolytic disorders are better understood, measurements of autohemolysis may be modified so as to enhance their usefulness in detecting and differentiating the various hemolytic states. It also seems likely that further studies on in vitro autohemolysis will help to elucidate some hemolytic mechanisms operating in vivo.

Receptor Mediated Binding of the Fibrinolytic Components, Plasminogen and Urokinase, to Peripheral Blood Cells

1987 ◽  
Vol 58 (03) ◽  
pp. 936-942 ◽  
Author(s):  
Lindsey A Miles ◽  
Edward F Plow
Keyword(s):  
T Cells ◽  
B Cell ◽  
Peripheral Blood ◽  
Binding Sites ◽  
Blood Cells ◽  
High Capacity ◽  
Red Cells ◽  
Peripheral Blood Cells ◽  
Cellular Functions ◽  
Fibrinolytic Proteins

SummaryGlu-plasminogen binds to platelets; the monocytoid line, U937, and the human fetal fibroblast line, GM1380 bind both plasminogen and its activator, urokinase. This study assesses the interaction of these fibrinolytic proteins with circulating human blood cells. Plasminogen bound minimally to red cells but bound saturably and reversibly to monocytes, granulocytes and lymphocytes with apparent Kd values of 0.9-1.4 μM. The interactions were of high capacity with 1.6 to 49 × 105 sites/cell and involved the lysine binding sites of plasminogen. Both T cells and non-rosetting lymphocytes and two B cell lines saturably bound plasminogen. Urokinase bound saturably to gianulocytes, monocytes, non-rosetting lymphocytes and a B cell line, but minimally to T cells, platelets and red cells. Therefore, plasminogen binding sites of high capacity, of similar affinities, and with common recognition specificities are expressed by many peripheral blood cells. Urokinase receptors are also widely distributed, but less so than plasminogen binding sites. The binding ol plasminogen and/ or urokinase to these cells may lead to generation of cell- associated proteolytic activity which contributes to a variety of cellular functions.

scholarly journals Determinants of Erythrocyte Size in Chronic Liver Disease

Blood ◽  
1969 ◽  
Vol 34 (6) ◽  
pp. 739-746 ◽  
Author(s):  
THOMAS M. KILBRIDGE ◽  
PAUL HELLER
Keyword(s):  
Bone Marrow ◽  
Liver Disease ◽  
Chronic Liver Disease ◽  
Peripheral Blood ◽  
Hepatic Cirrhosis ◽  
Alcoholic Cirrhosis ◽  
Clinical Findings ◽  
Red Cells ◽  
Red Cell ◽  
Cell Volumes

Abstract Serial determinations of red cell volumes were made with an electronic sizing device in 30 patients with hepatic cirrhosis. Variations in red cell volumes were correlated with other hematologic and clinical findings. The results of these studies suggest that volume macrocytosis in patients with alcoholic cirrhosis is either due to megaloblastosis of the bone marrow or to an accelerated influx of young red cells into the peripheral blood.

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