scholarly journals THE PHYSICAL STATE OF HEMOGLOBIN IN SICKLE-CELL ANEMIA ERYTHROCYTES IN VIVO

1968 ◽  
Vol 127 (4) ◽  
pp. 711-716 ◽  
Author(s):  
Johanna Döbler ◽  
John F. Bertles
Keyword(s):  
Electron Microscopy ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Physical State ◽  
Venous Blood ◽  
Significant Loss ◽  
Filamentous Form ◽  
Hb S

Venous blood removed anaerobically from patients with sickle-cell anemia was transferred immediately into fixative, thus precluding significant loss or gain of oxygen by the cells. Electron microscopy demonstrated an intraerythrocytic fibrillar fine structure similar to that described in prior studies on erythrocytes sickled by deoxygenation in vitro. Observations reported here lead to these conclusions: (a) explanations of the sickling process derived from in vitro experimentation may with validity be applied to sickling in vivo; and (b) the term "sickled" must be used with caution: a sickle-shaped membrane does not necessarily endose Hb S in filamentous form.

scholarly journals Sickle Cell Anemia and Babesia Infection

Pathogens ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 1435
Author(s):  
Divya Beri ◽  
Manpreet Singh ◽  
Marilis Rodriguez ◽  
Karina Yazdanbakhsh ◽  
Cheryl Ann Lobo
Keyword(s):  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Current Knowledge ◽  
Globin Gene ◽  
Environmental Niche ◽  
Parasite Resistance ◽  
The Usa ◽  
Human Infections

Babesia is an intraerythrocytic, obligate Apicomplexan parasite that has, in the last century, been implicated in human infections via zoonosis and is now widespread, especially in parts of the USA and Europe. It is naturally transmitted by the bite of a tick, but transfused blood from infected donors has also proven to be a major source of transmission. When infected, most humans are clinically asymptomatic, but the parasite can prove to be lethal when it infects immunocompromised individuals. Hemolysis and anemia are two common symptoms that accompany many infectious diseases, and this is particularly true of parasitic diseases that target red cells. Clinically, this becomes an acute problem for subjects who are prone to hemolysis and depend on frequent transfusions, like patients with sickle cell anemia or thalassemia. Little is known about Babesia’s pathogenesis in these hemoglobinopathies, and most parallels are drawn from its evolutionarily related Plasmodium parasite which shares the same environmental niche, the RBCs, in the human host. In vitro as well as in vivo Babesia-infected mouse sickle cell disease (SCD) models support the inhibition of intra-erythrocytic parasite proliferation, but mechanisms driving the protection of such hemoglobinopathies against infection are not fully studied. This review provides an overview of our current knowledge of Babesia infection and hemoglobinopathies, focusing on possible mechanisms behind this parasite resistance and the clinical repercussions faced by Babesia-infected human hosts harboring mutations in their globin gene.

scholarly journals Sickling Cells, Cyclic Nucleotides, and Protein Kinases: The Pathophysiology of Urogenital Disorders in Sickle Cell Anemia

Anemia ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
Mário Angelo Claudino ◽  
Kleber Yotsumoto Fertrin
Keyword(s):  
Protein Kinases ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Cyclic Nucleotides ◽  
Single Point ◽  
Clinical Manifestations ◽  
Single Point Mutation ◽  
Tract Infections

Sickle cell anemia is one of the best studied inherited diseases, and despite being caused by a single point mutation in theHBBgene, multiple pleiotropic effects of the abnormal hemoglobin S production range from vaso-occlusive crisis, stroke, and pulmonary hypertension to osteonecrosis and leg ulcers. Urogenital function is not spared, and although priapism is most frequently remembered, other related clinical manifestations have been described, such as nocturia, enuresis, increased frequence of lower urinary tract infections, urinary incontinence, hypogonadism, and testicular infarction. Studies on sickle cell vaso-occlusion and priapism using bothin vitroandin vivomodels have shed light on the pathogenesis of some of these events. The authors review what is known about the deleterious effects of sickling on the genitourinary tract and how the role of cyclic nucleotides signaling and protein kinases may help understand the pathophysiology underlying these manifestations and develop novel therapies in the setting of urogenital disorders in sickle cell disease.

scholarly journals The effect of fixed oil extracts of Nigella sativa on sickle cells: An in-vitro study in Khartoum state -Sudan

2021 ◽  
Vol 10 (3) ◽  
pp. 317-321
Author(s):  
Eman Ali Abuagla Dafaalla ◽  
Amira Ahmed Khalid Humeida
Keyword(s):  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Antioxidant Activities ◽  
Nigella Sativa ◽  
Venous Blood ◽  
In Vitro Study ◽  
Hemoglobin Concentration ◽  
Inherited Disorders ◽  
P Values

In Sudan, sickle cell anemia (SCA) is one of the most common inherited disorders of hemoglobin that has complications and becomes world problem. Nigella sativa NS (black seed) has been reported to have calcium antagonist and antioxidant activities, both of which play a role in the management of sickle cell anemia. The aim of the current study was to evaluate of the anti-sickling activity of the NS extracts, forty patients with sickle cell anemia were recruited for the study. A total of 3ml of venous blood was collected from each patient after obtaining the consent and the ethical approval, the blood was treated with Nigella sativa (NS) extract, and sickling test was performed. Descriptive study and p-values were used, and the correlation was evaluated, data was analyzed by SPSS. Recent study found that the sickling test after treating blood with Nigella sativa extract showed a negative result in 75% of patients and a 25% of patients showed persistently positive sickling test result. The anti-sickling effect in relation to various hemoglobin concentration, gender and age group showed p values of 0.007, 0.672 and 0.853 respectively indicating significant relationship with Hb concentration. Our study concluded that the fixed oil extract from Nigella sativa has an in-vitro anti-sickling activity on patients with SCA and this finding could indicate the use of this extract in treatment.

scholarly journals Abnormal Cytokine Production By Mast Cell Cultures from Sickle Cell Anemia Patients in Response to Inflammatory Stimuli and to Co-Culture with Eosinophils

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 3566-3566
Author(s):  
Myriam Salazar-Terreros ◽  
Kleber Yotsumoto Fertrin ◽  
Nicolas Moreno Reyes ◽  
Fernando Ferreira Costa ◽  
Carla F. Franco-Penteado
Keyword(s):  
Mast Cell ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Peripheral Blood ◽  
Cytokine Production ◽  
Fold Increase ◽  
Individual Response ◽  
Activated State

Mast cell function (MC) in pathologic states can be studied through their ability to secrete mediators in vitro depending on MC phenotype and the nature of the stimuli. Reports on MC mediators in sickle cell anemia (SCA) patients are scarce, but clinical signs of MC activation syndrome, such as increased plasma histamine in vaso‐occlusive crisis (VOC), and normal or slightly elevated serum tryptase have been reported. However, assessing the biological relevance of MC as a cytokine source is more challenging because it is unclear under which circumstances they secrete those products in vivo, or if the cytokines measured systemically stem from a different cell type. We aimed to investigate the profile of mediators involved in the inflammatory process produced by MC in SCA. Methods:The supernatant of 5-week old MC cultures (17 SCA, 8 HV) obtained from peripheral blood CD34+ cells from 29 SCA patients and 13 healthy volunteers (HV) was analyzed using a multiplex platform and colorimetric assays for endothelin-1 (ET-1) and substance P (SP) (10 SCA, 6 HV). A correlation matrix (Pearson correlations, R software, v. 3.6.1) was generated using laboratory and clinical data chosen based on their value as inflammatory or prognosis markers (hydroxyurea [HU] treatment, fetal hemoglobin [HbF], hemoglobin [Hb], vaso-occlusive crisis [VOC], percentage of peripheral blood neutrophils, eosinophils (Eos), basophils, erythroblasts, and reticulocytes), MC surface expression of CD117, CD48 and CD63, and the supernatant content of 11 cytokines. To investigate MC cytokine release, we tested the supernatants from Eos-MC co-cultures (3:1 ratio), and after stimulation with ET-1 (20 nM), SP (10 µM) and imatinib (20 µg/ml)(n=3 per treatment). Results: Out of 26 cytokines, we found elevated levels of the following in the supernatants of SCA-MC cultures (data represented as mean in pg/ml±SE): TNFα: SCA=88.7±18.4, HV=32.6±3.8; IFNγ: SCA=55.3±11.2, HV=15.7±1.8; MCP1: SCA=555.0±147.2, HV=145.3±35.2; RANTES: SCA=24.7±3.9, HV=10.7±1.8 (p<0.05). However, SCA-MC from patients treated with HU (n = 11) showed higher values of IL-1b, IL-4, IL-5, IL-9, IL-15, and FGF than HV (n=8) and HU-free patients (n=6) (p<0.05). Supernatants from SCA-MC had higher ET-1 production compared to HV-MC (SCA=16.3±1.2, HV=11.93±1.3, pg/ml, p=0.02) but SP production was similar (SCA=27.9±1.3; HV=31.49±0.7 pg/ml). ET-1 stimulation of MC cultures caused 2-fold increase in IL-1AR production on HV-MC, but failed to produce any effect on SCA-MC. Similarly, imatinib reduced FGF only in HV-MC samples (HV: 15.1±3.5, HV-HU: 4.0±1.6, pg/ml). No effect on cytokine production was observed with SP. Conversely, Eos-MC cocultures showed a 10- and 4-fold increase of IL-5 and IL-9, respectively, regardless of the origin of Eos (HV or SCA). SCA-MC/SCA-Eos co-cultures had elevated proinflammatory (IL-1b, IL-12, TNF-α) and angiogenic (FGF, VEGF) cytokines, RANTES, IL-7, IL-4, and IL1-RA compared with SCA-MC/HV-Eos and HV-MC/HV-Eos (p<0.05). Preliminary multiparametric analysis on data from SCA patients showed a strong negative correlation between HU therapy and VGEF production, and between HbF levels and CD63 expression (MC activation marker).We also found a positive correlation between history of VOC and eotaxin-1 produced by SCA-MC. Conclusions: We found that MC responses depend both on the origin of the cultured cell and the stimuli utilized. Despite differences between in vitro and in vivo MC populations, our data show that cultured SCA-MC have a sustained activated state and produce a repertoire of mediators that could contribute to a perivascular microenvironment in favor of leukocyte and endothelium activation. In terms of cytokine production, cultured SCA-MC were more sensitive to stimulation by SCA-Eos than by HV-Eos, which may be relevant to the pathophysiology of airway inflammation in SCA patients with asthma. Differences in cytokine production between SCA-MC cultures from patients treated or not with HU may reflect the variability in adherence to treatment, individual response to each compound, or epigenetic modifications during the MC differentiation process that affect the phenotype of the mature MC. These results support that mediators produced by MC can contribute to the chronic inflammatory state and may be implicated in exacerbated responses to eosinophil activation in SCA. Disclosures Fertrin: Agios Pharmaceuticals, Inc.: Research Funding.

scholarly journals Metabolomic Characterization of Red Blood Cell Differentiation

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 35-35
Author(s):  
Kelsey Temprine ◽  
Amanda Sankar ◽  
Costas Lyssiotis ◽  
Yatrik Shah
Keyword(s):  
Amino Acid ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Sodium Butyrate ◽  
Global Changes ◽  
Ineffective Erythropoiesis ◽  
Irreversible Loss ◽  
In Vivo Models

Background: Erythropoiesis is the highly coordinated multi-step process by which multipotent hematopoietic stem cells differentiate into mature enucleated red blood cells (RBCs). As erythroid cells become more terminally differentiated, they undergo changes in morphology and gene expression, start synthesizing hemoglobin, commit to an irreversible loss of proliferation, and eventually expulse their nuclei and other cytoplasmic organelles. Thus, RBCs must rely on their proteome and metabolome for proper function. The RBC proteome is estimated to contain 2,800 proteins, including a variety of receptors and transporters that allow RBCs to uptake xenobiotics or endogenous metabolites as they circulate for ∼120 days. Furthermore, they are metabolically active with glycolysis, nucleotide catabolism, and glutathione metabolism as the major pathways supporting cell survival and function. However, it is unclear how the metabolome is altered during erythropoiesis, what role metabolites play in normal erythropoiesis, and if dysregulation of metabolites contributes to diseases of ineffective erythropoiesis, such as sickle cell anemia and thalassemia. Methods: Four models of erythropoiesis were used in this study. 1) Mice were treated with phenylhydrazine (Phz) to induce acute hemolysis followed by erythropoietic recovery, leading to an increase in circulating reticulocytes. 2) Mice were lethally irradiated and transplanted with wild-type or sickle cell bone marrow, leading to anemic profiles in sickle cell chimeras. 3) The mouse erythroleukemic (MEL) cell line was treated with DMSO to induce differentiation. 4) The human erythroleukemic (K562) cell line was treated with sodium butyrate to induce differentiation. For the in vivo mouse models, blood was collected from control and treated animals, and complete blood count (CBC) analysis was performed. For the in vitro cell culture models, the mRNA levels of β-globin were measured by Q-RT-PCR in control and differentiated cells, and the degree of hemoglobinization was determined visually and via staining for heme. In addition, metabolites were extracted from the collected RBCs and erythroleukemic cell lines, and a Snapshot LC/MS metabolomic platform was used to identify commonly altered metabolites. Results: We first validated our four models of erythropoiesis. Treatment with Phz decreased the number of total RBCs while increasing the RBC distribution width, indicating an increased number of reticulocytes (more immature RBCs) in circulation. Similar results were seen in the sickle cell chimeras. Treatment of MEL and K562 cells with DMSO and sodium butyrate, respectively, resulted in increased expression of β-globin, increased levels of heme, and increased red color. Then, using our Snapshot metabolomic platform, we identified global changes in RBC metabolism during erythropoiesis. Analyses of the commonly altered metabolites in the in vitro and in vivo models revealed an increase in amino acid, mitochondrial, and urea cycle metabolism during erythropoiesis. L-aspartate levels were particularly upregulated, especially in DMSO-treated MEL cells. We are now investigating the role of aspartate in the regulation of erythropoiesis. Conclusions: We defined how the metabolome was altered in multiple in vitro and in vivo models of erythropoiesis and identified global changes in RBC metabolism between the different models. Specifically, we found that L-aspartate was upregulated during RBC differentiation in all four models. Aspartate is an amino acid that plays a role in many processes in cells, including nucleotide biosynthesis, redox homeostasis, and amino acid biosynthesis. We hypothesize that aspartate metabolism is critical for RBC differentiation and that its dysregulation exacerbates disease of ineffective erythropoiesis, such as sickle cell anemia and β-thalassemia. We are currently testing its role in inducing hemoglobinization and in regulating the commitment of erythroid progenitor cells to an irreversible loss of proliferation. Overall, we believe that understanding the precise mechanisms by which cellular metabolism plays a role in proper RBC differentiation may lead to better therapies for diseases of ineffective erythropoiesis, such as sickle cell anemia and thalassemia. Disclosures No relevant conflicts of interest to declare.

scholarly journals A Transgenic Mouse Model of Hemoglobin S Antilles Disease

Blood ◽  
1997 ◽  
Vol 89 (11) ◽  
pp. 4204-4212 ◽  
Author(s):  
R.A. Popp ◽  
D.M. Popp ◽  
S.G. Shinpock ◽  
M.Y. Yang ◽  
J.G. Mural ◽  
...  
Keyword(s):  
Sickle Cell ◽  
Experimental Studies ◽  
Oxygen Affinity ◽  
High Oxygen ◽  
Lower Solubility ◽  
Lower Oxygen ◽  
Hb S ◽  
High Oxygen Affinity

Abstract Hemoglobin (Hb) S Antilles is a naturally occurring form of sickling human Hb but causes a more severe phenotype than Hb S. Two homozygous viable Hb S Antilles transgene insertions from Tg58Ru and Tg98Ru mice were bred into MHOAH mice that express high oxygen affinity (P50 ∼24.5 mm Hg) rather than normal (P50 ∼40 mm Hg) mouse Hbs. The rationale was that the high oxygen affinity MHOAH Hb, the lower oxygen affinity of Hb S Antilles than Hb S (P50 ∼40 v 26.5 mm Hg), and the lower solubility of deoxygenated Hb S Antilles than Hb S (∼11 v 18 g/dL) would favor deoxygenation and polymerization of human Hb S Antilles in MHOAH mouse red blood cells (RBCs). The Tg58 × Tg98 mice produced have a high and balanced expression (∼50% each) of hα and hβS Antilles globins, 25% to 35% of their RBCs are misshapen in vivo, and in vitro deoxygenation of their blood induces 30% to 50% of the RBCs to form classical looking, elongated sickle cells with pointed ends. Tg58 × Tg98 mice exhibit reticulocytosis, an elevated white blood cell count and lung and kidney pathology commonly found in sickle cell patients, which should make these mice useful for experimental studies on possible therapeutic intervention of sickle cell disease.

scholarly journals Prolongation of sickle cell survival by dimethyl adipimidate is compromised by immune sensitization

Blood ◽  
1984 ◽  
Vol 64 (1) ◽  
pp. 161-165 ◽  
Author(s):  
MS Guis ◽  
WM Lande ◽  
N Mohandas ◽  
R Pennathur-Das ◽  
H Preisler ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Initial Study ◽  
Red Cells ◽  
Adult Males ◽  
Cell Disease ◽  
Sickle Cells

Abstract The effect of dimethyl adipimidate (DMA), an amino-reactive crosslinking reagent with demonstrated antisickling properties in vitro, on the survival of 51Cr-labeled autologous sickle cells was evaluated in five adult males with sickle cell anemia. The survival of cells pretreated with 5 mmol/L DMA (pH 7.4), normal (t1/2 28–33 days) in four subjects and near-normal (t1/2 20 days) in the fifth, was considerably longer than that usually observed in sickle cell disease. In fact, the effect of DMA on the survival of sickle cells in vivo equals or exceeds that of any other agent tested to date. In three subjects, the survival of a second infusion of DMA-treated red cells was much shorter (t1/2 1.8, 3, 4.5 days) than in the initial study. An antibody was detected in the serum of these subjects that was directed to DMA-treated red cells. Modification of the immunogenicity of treated cells will be required before further consideration of DMA for use in the therapy of sickle cell anemia.

scholarly journals Regulation of Fetal and Adult Hemoglobin Formation in Patients with Sickle Cell Disease Transfused to Normal Hematocrits

Blood ◽  
1967 ◽  
Vol 29 (1) ◽  
pp. 126-133 ◽  
Author(s):  
THOMAS G. GABUZDA ◽  
FRANK H. GARDNER ◽  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Control Mechanisms ◽  
Cell Disease ◽  
Short Term ◽  
Hemoglobin Synthesis ◽  
Hb S ◽  
Adult Hemoglobin

Abstract Two patients with sickle cell anemia and one patient with sickle thalassemia were acutely transfused to normal hemoglobin levels. The synthesis of fetal and adult (in this instance, Hb S) hemoglobins was assessed in cohorts of reticulocytes emerging during the period of erythroid suppression which followed. In two patients observations were also made during the period of recovery from the suppression. Hemoglobin synthesis in the reticulocytes was assessed by means of in vitro incorporation of C14-labeled leucine into the separate hemoglobins. The results in all three patients suggested that both fetal and adult hemoglobin synthesis are under similar physiologic control mechanisms during short-term alterations in the state of erythropoiesis; the formation of both was almost completely stopped soon after transfusion, and both were almost simultaneously reactivated as the patients returned to the anemic state.

scholarly journals Prolongation of sickle cell survival by dimethyl adipimidate is compromised by immune sensitization

Blood ◽  
1984 ◽  
Vol 64 (1) ◽  
pp. 161-165
Author(s):  
MS Guis ◽  
WM Lande ◽  
N Mohandas ◽  
R Pennathur-Das ◽  
H Preisler ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Initial Study ◽  
Red Cells ◽  
Adult Males ◽  
Cell Disease ◽  
Sickle Cells

The effect of dimethyl adipimidate (DMA), an amino-reactive crosslinking reagent with demonstrated antisickling properties in vitro, on the survival of 51Cr-labeled autologous sickle cells was evaluated in five adult males with sickle cell anemia. The survival of cells pretreated with 5 mmol/L DMA (pH 7.4), normal (t1/2 28–33 days) in four subjects and near-normal (t1/2 20 days) in the fifth, was considerably longer than that usually observed in sickle cell disease. In fact, the effect of DMA on the survival of sickle cells in vivo equals or exceeds that of any other agent tested to date. In three subjects, the survival of a second infusion of DMA-treated red cells was much shorter (t1/2 1.8, 3, 4.5 days) than in the initial study. An antibody was detected in the serum of these subjects that was directed to DMA-treated red cells. Modification of the immunogenicity of treated cells will be required before further consideration of DMA for use in the therapy of sickle cell anemia.

Various Drugs with or without an Antisickling Effect in the In Vitro Tests Showed a Strong Antisickling Effect in the In Vivo Studies in Transgenic Sickle Mice.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 3190-3190
Author(s):  
Toshio Asakura ◽  
Osheiza Abdulmalik ◽  
Efe Iyamu ◽  
Qiukan Chen ◽  
Jisheng Yang ◽  
...  
Keyword(s):  
Adverse Effects ◽  
Sickle Cell ◽  
In Vivo Studies ◽  
In Vitro Tests ◽  
Reference Laboratory ◽  
Beneficial Effects ◽  
Combined Use ◽  
Hb S

Abstract Since its inception in 1997, the NHLBI Sickle Cell Disease (SCD) Reference Laboratory (Ref Lab) has received over 150 candidate drugs from over 30 researchers, universities and companies for evaluation. All candidate drugs are evaluated for both their beneficial and adverse effects in vitro using more than 10 sophisticated methods. Specifically, each candidate drug is studied to see whether it inhibits cell sickling, prolongs the delay time prior to deoxy-hemoglobin (Hb) S polymerization, increases the solubility of deoxy-Hb S, hydrates red blood cells (RBCs), prevents dehydration of RBCs, shifts the oxygen equilibrium curve (OEC) toward the left, reduces the adhesion of RBCs to endothelial cells, promotes the synthesis of Hb F, causes hemolysis, promotes formation of met-Hb and increases the denaturation of intracellular Hb S. With this new approach, we have found more than 10 new antisickling agents that showed beneficial effects without significant adverse effects in the in vitro tests. Of these, five drugs were further studied in vivo using transgenic mice that produce human Hb S, and all five drugs inhibited the formation of sickled cells in the blood and significantly prolonged their survival time under acute hypoxic conditions. The five drugs are NIPRISAN (plant extracts) [Brit. J. Haematol. (BJH) 122:1001,2003], MX-1520 (prodrug of vanillin) (BJH125: 788,2004), 5-hydroxymethyl-2-furfural (5HMF) (BJH128:552,2005), NS3623 (Blood 97: 1461,2001, in vivo results not shown), and FLOCOR (surfactant) (Blood94:420a,1999). Of these 5 drugs, FLOCOR and NS3623 did not have an antisickling effect in vitro. It is quite interesting that FLOCOR and NS3623 which showed no direct antisickling effect in vitro, showed a strong antisickling effect in vivo. Some of our results suggest that combined use of some of these drugs may have a synergistic effect in vivo. Studies on the combined use of these drugs are planned. Researchers with candidate drugs that may have beneficial effects for SCD are welcome to contact the SCD Ref Lab for free in vitro evaluation of their agents (for more information, please see our website at www.tatcom.com/sickle-cell).

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