Anti-Apoptotic Effect, Mediated by Elevated Intracellular Cyclic AMP Levels, in Neutrophils of Sickle Cell Disease Patients.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2343-2343
Author(s):  
Nicola Conran ◽  
Carolina Lanaro ◽  
Fernanda G. Pereira ◽  
Tohru Ikuta ◽  
Sara T.O. Saad ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Autologous Serum ◽  
Apoptotic Cells ◽  
Cell Disease ◽  
Survival Factors ◽  
Chronic Inflammatory Condition ◽  
Control Serum ◽  
Apoptotic Effect ◽  
Inflammatory State

Abstract Sickle cell disease (SCD) is now widely regarded as a chronic inflammatory condition, characterized by high base-line leukocyte counts, alterations in inflammatory cytokine levels, vascular endothelial injury and increased red and white cell adhesiveness. We hypothesized that this inflammatory state may be exacerbated by alterations in the apoptotic process in inflammatory cells, such as neutrophils. Neutrophils were isolated from the peripheral blood of healthy controls and individuals with SCD in steady state by separation over a ficoll-paque gradient. Following washing and lysis of contaminating red cells, neutrophils were cultured in DMEM supplemented with 10% (v/v) autologous serum and antibiotics for 20 h (4 x 106 cells/ml, 37oC, 5% CO2). Apoptosis was assessed by flow cytometry using a fluorescein isocyanate-labeled recombinant annexin V antibody and propidium iodide staining. Morphological analysis confirmed the apoptotic state of cells. After 20 h in culture, the percentage of non-apoptotic neutrophils from SCD patients (SCD neutrophils; 13.34 ± 1.03 %, n=8) was significantly higher than the percentage of non-apoptotic normal neutrophils (7.68 ± 1.59, n=7; P<0.01). Apoptosis is mediated by a number of signaling pathways, with the cAMP-dependent pathway being known to have an important anti-apoptotic role in neutrophils. Accordingly, measurement of cAMP in isolated normal and SCD neutrophils by ELISA demonstrated levels of cAMP to be significantly increased in the neutrophils of SCD individuals (4.55 ± 0.38 pMol/1x 106 cells compared to 2.18 ± 0.39 pMol/1x 106 cells in normal individuals, n>14, P<0.001). Co-incubation of SCD neutrophils during 20h-culture with a cAMP-dependent protein kinase (PKA) inhibitor, KT5720 (3μM), significantly decreased the percentage of non-apoptotic cells (12.88 ± 1.49 %, decreased to 6.18 ± 0.51 %, n=6; p=0.01) to levels similar to those seen in normal neutrophil cultures, indicating that the anti-apoptotic effect seen in SCD neutrophils is probably mediated by a cAMP-PKA dependent mechanism. Interestingly, when SCD neutrophils were cultured in medium containing fetal calf serum instead of autologous serum, the number of non-apoptotic cells at 20 h of culture was not different to the number of normal non-apoptotic neutrophils (8.91 ± 2.55 % compared to 9.08 ± 2.11 %, respectively, n= 7, P>0.05). This finding may indicate that this anti-apoptotic effect may be maintained by survival factors contained in serum; indeed incubation of normal neutrophils with SCD patient serum (10 % v/v, 30 min, 37oC), but not control serum, significantly augmented intracellular cAMP levels by 117.9 ± 10.5 % (n=8, p<0.01). In conclusion, we have observed an increased survival of neutrophils from SCD individuals under culture conditions, probably mediated by an up-regulated cAMP-PKA pathway and possibly dependent on cAMP-elevating survival factors contained in the serum. Since neutrophil apoptosis is imperative for the resolution of inflammation, increased neutrophil survival may be an important contributing factor to the chronic inflammatory state that characterizes SCD.

HMGB1 Expression in Sickle Cell Disease: A Pro-Inflammatory Cytokine and Potential Therapeutic Target.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3803-3803
Author(s):  
Myra F. Barginear ◽  
Anu G. Chandok ◽  
Cristina Sison ◽  
Lihong Yang ◽  
Daniel R. Budman ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Therapeutic Target ◽  
Vascular Endothelial Cells ◽  
Therapeutic Benefit ◽  
Cell Disease ◽  
Extracellular Milieu ◽  
Hmgb1 Expression ◽  
Attractive Therapeutic Target ◽  
Inflammatory State

Abstract Background: HMGB1 is traditionally known as a DNA binding protein involved in gene transcription. When released into the extracellular milieu, HMGB1 activates vascular endothelial cells and macrophages/monocytes to express proinflammatory cytokines, chemokines, and adhesion molecules. Therefore, HMGB1 may be a good marker for disease activity in Sickle Cell Disease (SCD), a chronic inflammatory state. Methods: After informed consent as per Institutional guidelines, plasma was obtained, frozen, and batch run for HMGB1 levels by ELISA technique in patients with SCD during non-crisis and crisis states. The detection limit of HMGB1 was achieved at concentrations above 1 ng/ml. Each experiment was repeated at least five times to ensure reproducibility. Results: 12 patients enrolled, 6 patients have gone into crisis. HMGB1 levels were measured on consecutive days for 5 five days during a crisis. We established mean HMGB1 expression levels of 32.83 ± 27.13ng/ml during a non-crisis state and 52.57± 15.22 ng/ml during a crisis, respectively (p< .028). In noninflammatory states, HMGB1 is < 5 ng/ml. Conclusion: We have demonstrated, for the first time, a persistent expression of HMGB1 in SCD patients. Although recruitment is on going, this data is consistent with pre-clinical data indicating that HMGB1 may be an important diffusible signal of necrosis-mediated inflammation. Additionally, our laboratory has previously demonstrated therapeutic benefit by neutralization of HMGB1 in sepsis. Thus if confirmed in a larger cohort, anti-HMGB1 therapy to reduce the effects of inflammation may be an attractive therapeutic target in sickle cell disease.

scholarly journals Red blood cell alloimmunization is influenced by recipient inflammatory state at time of transfusion in patients with sickle cell disease

2014 ◽  
Vol 168 (2) ◽  
pp. 291-300 ◽  
Author(s):  
Ross M. Fasano ◽  
Garrett S. Booth ◽  
Megan Miles ◽  
Liping Du ◽  
Tatsuki Koyama ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Blood Cell ◽  
Sickle Cell ◽  
Red Blood Cell ◽  
Cell Disease ◽  
Inflammatory State

scholarly journals Circulating Microparticles in Children With Sickle Cell Anemia in a Tertiary Center in Upper Egypt

2019 ◽  
Vol 25 ◽  
pp. 107602961982883 ◽  
Author(s):  
Asmaa M. Zahran ◽  
Khalid I. Elsayh ◽  
Khaled Saad ◽  
Mostafa M. Embaby ◽  
Mervat A. M. Youssef ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Pediatric Patients ◽  
Healthy Children ◽  
Cell Disease ◽  
Thrombotic Risk ◽  
Chronic Inflammatory Condition ◽  
Tertiary Center ◽  
Circulating Microparticles

Sickle cell disease (SCD) is a genetically inherited hemolytic anemia increasingly appreciated as a chronic inflammatory condition and hypercoagulable state with high thrombotic risk. It is associated with disturbed immune phenotype and function and circulating microparticles (MPs) derived from multiple cell sources. This study was carried out to determine MPs profiles in patients with sickle cell anemia (either on hydroxyurea (HU) therapy or those with no disease-modifying therapy) and to compare these profiles with healthy children. Moreover, our study assesses the potential impact of HU on other aspects of circulating MPs. We performed a cross-sectional study on 30 pediatric patients with SCD divided by treatment into 2 groups (those receiving HU or no therapy) attending Hematology Clinic and 20 age-matched healthy children. The blood samples obtained were analyzed for MPs by flow cytometry. Sickle cell disease group with no therapy showed elevated levels of total, platelet, and erythroid MPs. In contrast, therapy with HU was associated with normalization of MPs. This study provided additional evidence that HU is an effective treatment option in pediatric patients with SCD, as it seems that it decreases the abnormally elevated MPs in those patients.

scholarly journals Loss of Bone in Sickle Cell Trait and Sickle Cell Disease Female Mice Is Associated With Reduced IGF-1 in Bone and Serum

Endocrinology ◽  
2016 ◽  
Vol 157 (8) ◽  
pp. 3036-3046 ◽  
Author(s):  
Liping Xiao ◽  
Biree Andemariam ◽  
Pam Taxel ◽  
Douglas J. Adams ◽  
William T. Zempsky ◽  
...  
Keyword(s):  
Sickle Cell Disease ◽  
Bone Formation ◽  
Sickle Cell ◽  
Bone Mineral ◽  
Sickle Cell Trait ◽  
Femoral Diaphysis ◽  
Cell Disease ◽  
Mineral Density ◽  
Control Serum

Characterization of the bone phenotype of 24-week-old female transgenic sickle cell disease (SCD), sickle cell trait (SCT) revealed significant reductions in bone mineral density and bone mineral content relative to control with a further significant decreased in SCD compared with SCT. By microcomputed tomography, femur middiaphyseal cortical area was significantly reduced in SCT and SCD. Cortical thickness was significantly decreased in SCD vs control. Diaphysis structural stiffness and strength were significantly reduced in SCT and SCD. Histomorphometry showed a significant increase in osteoclast perimeter in SCD and significantly decreased bone formation in SCD and SCT compared with control with a further significant decrease in SCD compared with SCT. Collagen-I mRNA was significantly decreased in tibiae from SCT and SCD and osterix, Runx2, osteoclacin, and Dmp-1 mRNA were significantly decreased in tibiae of SCD compared with control. Serum osteocalcin was significantly decreased and ferritin was significantly increased in SCD compared with control. Igf1 mRNA and serum IGF1 were significantly decreased in SCD and SCT. IGF1 protein was decreased in bone marrow stromal cells from SCT and SCD cultured in osteogenic media. Crystal violet staining revealed fewer cells and significantly reduced alkaline phosphatase positive mineralized nodules in SCT and SCD that was rescued by IGF1 treatment. We conclude that reduced bone mass in SCD and SCT mice carries architectural consequences that are detrimental to the mechanical integrity of femoral diaphysis. Furthermore reduced IGF1 and osteoblast terminal differentiation contributed to reduced bone formation in SCT and SCD mice.

Sickle blood contains tissue factor–positive microparticles derived from endothelial cells and monocytes

Blood ◽  
2003 ◽  
Vol 102 (7) ◽  
pp. 2678-2683 ◽  
Author(s):  
Arun S. Shet ◽  
Omer Aras ◽  
Kalpna Gupta ◽  
Mathew J. Hass ◽  
Douglas J. Rausch ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Steady State ◽  
Sickle Cell Disease ◽  
Tissue Factor ◽  
Sickle Cell ◽  
Endothelial Activation ◽  
Cell Disease ◽  
Control Subjects ◽  
Inflammatory State ◽  
And Control

Abstract Blood microparticles (MPs) in sickle cell disease (SCD) are reportedly derived only from erythrocytes and platelets. Yet in SCD, endothelial cells and monocytes are activated and abnormally express tissue factor (TF). Thus, sickle blood might contain TF-positive MPs derived from these cells. With the use of flow cytometry to enumerate and characterize MPs, we found total MPs to be elevated in crisis (P = .0001) and steady state (P = .02) in subjects with sickle cell disease versus control subjects. These MPs were derived from erythrocytes, platelets, monocytes, and endothelial cells. Erythrocyte-derived MPs were elevated in sickle crisis (P = .0001) and steady state (P = .02) versus control subjects, as were monocyte-derived MPs (P = .0004 and P = .009, respectively). Endothelial and platelet-derived MPs were elevated in sickle crisis versus control subjects. Total TF-positive MPs were elevated in sickle crisis versus steady state (P = .004) and control subjects (P < .0001) and were derived from both monocytes and endothelial cells. Sickle MPs shortened plasma-clotting time compared with control MPs, and a TF antibody partially inhibited this procoagulant activity. Markers of coagulation were elevated in patients with sickle cell disease versus control subjects and correlated with total MPs and TF-positive MPs (P < .01 for both). These data support the concept that SCD is an inflammatory state with monocyte and endothelial activation and abnormal TF activity. (Blood. 2003;102:2678-2683)

scholarly journals Higher Prevalence of Autoimmune Diseases in Patients with Sickle Cell Disease

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 982-982
Author(s):  
Man Wai Tang ◽  
Erfan Nur ◽  
Charlotte F.J. Van Tuijn ◽  
Bart J. Biemond
Keyword(s):  
Sickle Cell Disease ◽  
General Population ◽  
Autoimmune Diseases ◽  
Chronic Inflammation ◽  
Sickle Cell ◽  
Research Funding ◽  
Patient Characteristics ◽  
Organ Damage ◽  
Cell Disease ◽  
Inflammatory State

Abstract Introduction Sickle cell disease (SCD) affects millions of people throughout the world. This is caused by a mutation in the hemoglobin gene resulting in abnormal red blood cells. Patients with SCD are in a state of chronic inflammation that is driven by ongoing hemolysis and ischemia-reperfusion injury due to recurrent vaso-occlusions.(1) It is also known that infections occur more frequently in patients with SCD. Abundant literature support the role of infections, interacting with the immune system as the so-called second hit in the cascade towards development of autoimmune diseases.(2) The chronic inflammation together with genetic predisposition and environmental factors can potentially lead to an auto-inflammatory state and/or disease. Recently, a study has shown a prevalence of autoimmune diseases (AID) of 1,3% in SCD patients between 7 and 17 years of age, although not compared to the general population.(3) Our hypothesis is that due to the chronic inflammation the prevalence of autoimmune diseases in adult patients with SCD are much higher compared to the general population. The aim of the study was to evaluate the proportion of patients with and without an AID. Methods Between 2004 and 2021, patients with SCD aged above 18 were seen at the outpatient clinic at the Amsterdam University Medical Center in Amsterdam. We performed a retrospective study in adult SCD patients to assess the prevalence of AID in SCD. AID was defined as: disease with the presence of autoantibodies and/or auto-reactive lymphocytes becoming involved in inflammation, which develop pathological autoimmunity and finally leads to tissue damage. We have selected 35 most common autoimmune diseases. A total of 338 patients with SCD were eligible and included in the study. The patient characteristics were summarized in Table 1. The previously reported prevalence of the AID in the African-American population was used to compare the prevalence in our study cohort. In addition, risk factors for AID and relation with organ damage was analyzed. Results AID was diagnosed in 36/338 patients with SCD. The prevalence of AID in this cohort is 10,7% compared to 4.7% in the general population (see table 2). There was no difference in patient characteristics (age, sex, genotype) between the SCD patients with or without AID. The BMI was higher in the group of patients with autoimmune diseases, although not statistically significant. In patients with SCD, the most frequent (>1%) diagnosed AID were: sudden deafness (1.8%), hyper- and hypothyroidism (3%) and sarcoidosis (1.2%). With respect to organ damage, a significantly high rate of retinopathy was observed in SCD patients with AID as compared to SCD patients without AID (53% and 29% respectively, p=0.005). Furthermore, a trend towards more frequently microalbuminuria was found in SCD patients with AID 14/36 (39%) as compared to patients without AID 69/302 (23%). Conclusions This study showed for the first time a higher prevalence of autoimmune diseases in adult patients with SCD compared to previous reported in the general population. In patients with AID, a trend towards more microalbuminuria and significantly higher rate of retinopathy were observed. These findings support the hypothesis that the chronic inflammatory state in SCD patients may be related to the development of AID. Further research is needed to find strategies to target the chronic inflammatory state in order to prevent the development of AID. References 1. Nader E, Romana M, Connes P. The Red Blood Cell-Inflammation Vicious Circle in Sickle Cell Disease. Front Immunol. 2020;11:454. 2. Ercolini AM, Miller SD. The role of infections in autoimmune disease. Clin Exp Immunol. 2009;155(1):1-15. 3. Li-Thiao-Te V, Uettwiller F, Quartier P, Lacaille F, Bader-Meunier B, Brousse V, et al. Coexistent sickle-cell anemia and autoimmune disease in eight children: pitfalls and challenges. Pediatr Rheumatol Online J. 2018;16(1):5. Figure 1 Figure 1. Disclosures Nur: Celgene: Speakers Bureau; Roche: Speakers Bureau; Novartis: Research Funding, Speakers Bureau. Biemond: Global Blood Therapeutics: Honoraria, Research Funding, Speakers Bureau; Sanquin: Research Funding; CSL Behring: Honoraria; Novo Nordisk: Honoraria; Celgene: Honoraria; Novartis: Honoraria, Research Funding, Speakers Bureau.

scholarly journals Asthma Management in Sickle Cell Disease

2013 ◽  
Vol 2013 ◽  
pp. 1-12 ◽  
Author(s):  
Esteban Gomez ◽  
Claudia R. Morris
Keyword(s):  
Sickle Cell Disease ◽  
Sickle Cell ◽  
Early Recognition ◽  
Asthma Management ◽  
Standard Of Care ◽  
Pulmonary Complications ◽  
Acute Chest Syndrome ◽  
Cell Disease ◽  
Inflammatory State ◽  
Genetic And Environmental Factors

Asthma is a common comorbid factor in sickle cell disease (SCD). However, the incidence of asthma in SCD is much higher than expected compared to rates in the general population. Whether “asthma” in SCD is purely related to genetic and environmental factors or rather is the consequence of the underlying hemolytic and inflammatory state is a topic of recent debate. Regardless of the etiology, hypoxemia induced by bronchoconstriction and inflammation associated with asthma exacerbations will contribute to a cycle of sickling and subsequent complications of SCD. Recent studies confirm that asthma predisposes to complications of SCD such as pain crises, acute chest syndrome, and stroke and is associated with increased mortality. Early recognition and aggressive standard of care management of asthma may prevent serious pulmonary complications and reduce mortality. However, data regarding the management of asthma in SCD is very limited. Clinical trials are needed to evaluate the effectiveness of current asthma therapy in patients with SCD and coincident asthma, while mechanistic studies are needed to delineate the underlying pathophysiology.

scholarly journals Cytokines Polymorphisms and Relationship with Cytokine Expression in Sickle Cell Disease

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4910-4910 ◽  
Author(s):  
Simone CO Gilli ◽  
Fernando V Pericole ◽  
Bruno Deltreggia Benites ◽  
Lilian Castilho ◽  
Marcelo Addas-Carvalho ◽  
...  
Keyword(s):  
Steady State ◽  
Sickle Cell Disease ◽  
Inflammatory Cytokines ◽  
Sickle Cell ◽  
Conflicts Of Interest ◽  
Rflp Analysis ◽  
Cell Disease ◽  
Genotype Frequencies ◽  
Number Of Patients ◽  
Chronic Inflammatory Condition

Abstract Sickle cell disease (SCD) is a chronic inflammatory condition, even in steady state, as indicated by elevated levels of inflammatory cytokines and increased Th17 responses, compared with healthy controls. These inflammatory pathways may be directly regulated by genetic polymorphisms and could be associated to different outcomes of the disease. High levels of a number of different circulating cytokines were found in SCD patients in several studies, and changes in the cytokine balance in SCD patients are an important risk factor for the occurrence of clinical events. Moreover, inter-patient variations in cytokine levels could be attributed to gene polymorphisms. To investigate cytokine polymorphisms and their association with cytokines expression we evaluated the IL4 intron3 VNTR (genotypes 1.1, 1.2, 2.2, 2.3), IL4T590C/T, IL6174G/C and TNFA308A/G polymorphisms and their correlation with TGFB, IL-4, IL-6 and IL-10 expression in steady-state SCD patients. Methods. Fourth-nine patients (24 male and 25 female; 39.8 ± 9.59 years) with SCD and 28 (22 male and 6 female; 35.5 ± 10.2 years) healthy blood donors were evaluated. The polymorphisms were performed by PCR-RFLP analysis described as [individuals (genotype frequencies)] and the expression of TGFB, IL-4, IL-6 and IL-10 by q-PCR expressed as [median (max-min)]. Results. A higher frequency of 1.2, 2.2 and 2.3 genotypes was found in SCD patients compared with normal controls [34(0.69) vs 12 (0.44), P=0.03]; higher expression of IL-4 was found in the ones carrying the 1.1 genotype [0.31 (2.53-0.01) vs 0.05 (0.95-0.0), P=0.047] and although no differences were found in the IL4T590C/T, IL6174G/C and TNFA308A/G polymorphism frequencies, a significantly greater expression of TGFB, IL6 and IL10 was observed in the patients cohort compared to normal individuals [1.55 (9.02-0.0) vs 0.97 (5.46-0.0), P=0.019]; [0.18 (45.45-0.0) vs 0.0 (8.14-0.0), P=0.03] and [0.98 (22.84-0.0) vs 0.0 (9.95-0.0), P<0.001, respectively]. All the genotype frequencies are consistent with Hardy-Weinberg equilibrium. Conclusion: A unique distribution of IL-4 genotypes was observed in our cohort of patients and controls, probably related to the miscellaneous ethnic background of our population. The highest prevalence of the IL4intron3 polymorphism in sickle cell patients suggests a less secretory phenotype associated with increased expression of inflammatory cytokines. IL-4 plays an important role in tissue adhesion and inflammation, including induction of adhesion molecules on vascular endothelial cells and could be responsible for a more “inflammatory” phenotype. Despite the small number of patients enrolled, our study brings insights and new data regarding the deregulation in immune system affecting SCD patients and this information must be investigated in larger cohorts, and may help to better characterize individual variations in immune responses and new markers for disease morbidity. Disclosures No relevant conflicts of interest to declare.

scholarly journals Elevated Urinary Leukotriene E4 Levels Are Associated with Hospitalization for Pain in Children with Sickle Cell Disease.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3405-3405
Author(s):  
Jeanine E. Jennings ◽  
Thiruvamoor Ramkumar ◽  
Jingnan Mao ◽  
Jessica Boyd ◽  
Mario Castro ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Sickle Cell Disease ◽  
Incidence Rate ◽  
Sickle Cell ◽  
Blood Cells ◽  
Medical Records ◽  
White Blood Cells ◽  
Cell Disease ◽  
Inflammatory Stimuli ◽  
Inflammatory State

Abstract Introduction: Sickle cell disease (SCD) is associated with an inflammatory state. Luekotrienes are inflammatory mediators derived from arachidonic acid and produced by white blood cells in response to inflammatory stimuli. We tested two hypotheses among children with SCD: Baseline leukotriene levels are elevated in SCD when compared to controls children without SCD, Baseline leukotriene levels are associated with an increased incidence rate of hospitalization for pain. Methods: During routine clinical visits, baseline urinary leukotriene E4 (LTE4) levels were measured in children with SCD (cases) and age, and ethnic, matched children without SCD (controls). Medical records of the cases were reviewed to assess the frequency of hospitalization for pain within three years of entering the study. Results: LTE4 levels were obtained in 71 cases and 22 controls. LTE4 levels were higher in the cases compared to controls (median LTE4: 100 vs.57 pg/mg creatinine); P < 0.001. After adjustment for age and diagnosis of asthma, a greater incidence rate of hospitalization for pain was observed among children with SCD in the highest tertile group when compared to the lowest tertile group (114 vs. 52 episodes per 100 patient-years, P=0.038). Conclusion: LTE4 levels are elevated in children with SCD when compared to controls without SCD and are associated with an increased rate of hospitalizations for pain.

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