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Individuals with sickle cell disease (SCD) present both chronic and acute inflammatory events. The TGF-β pathway is known to play a role in immune response, angiogenesis, inflammation, hematopoiesis, vascular inflammation, and cell proliferation. Polymorphisms in the transforming growth factor-beta receptor 3 (TGFBR3) gene have been linked to several inflammatory diseases. This study investigated associations between two TGFBR3 haplotypes and classical laboratory parameters, as well as clinical manifestations, in SCD. We found that individuals with the GG haplotype presented higher levels of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), triglycerides, non-HDL cholesterol, total proteins, and globulin than individuals with non-GG haplotypes. In addition, the GG haplotype was associated with a previous history of pneumonia. Individuals with the CGG haplotype presented increased plateletcrit, TC, LDL-C levels, and non-HDL cholesterol. The CCG haplotype was also associated with a previous history of pneumonia. Our findings suggest that individuals with the GG and CGG haplotypes of TGFBR3 present important alterations in lipid profile.

Introduction. Clinical complications in sickle cell anemia (SCA) are heterogeneous and involve several molecules. It has been suggested that SCA individuals present a dyslipidemic phenotype and that lipid parameters are associated with severe clinical complications, such as pulmonary hypertension. We sought to investigate associations between lipid parameters and clinical manifestations, as well as other laboratory parameters in a population of pediatric SCA patients.Methods. Our cross-sectional evaluation included 126 SCA patients in steady state and who were not undergoing lipid-lowering therapy. Hematological and biochemical parameters were characterized, and previous clinical manifestations were investigated.Results. Total cholesterol and low-density lipoprotein cholesterol (LDL-C) levels were increased in patients with a previous history of pneumonia, which also positively correlated with HbS levels. Decreased LDL-C levels were also associated with leg ulcers and anemia. Elevated high-density lipoprotein cholesterol (HDL-C) levels were associated with pain crises, increased viscosity, and decreased hemolysis. Several studies have determined that lipids play a role in the vascular impairment seen in SCA, which was corroborated by our findings.Conclusions. In sum, our results suggest that total cholesterol, HDL-C, and LDL-C levels are associated with hemolysis and anemia markers and, most importantly, with clinical complications related to vasculopathy in SCA.

Instituto Gonc alo Moniz- Fundacao Oswaldo Cruz (IGM-FIOCRUZ) and the Hospital da Crianc a das Obras Sociais Irma Dulce (HCOSID) approved this study, with protocol number 0016.0.225.000-09.

Individuals with sickle cell anemia (SCA) present chronic anemia, hemolysis, an exacerbated inflammatory response, and heterogeneous clinical complications, which may be modulated by the transforming growth factor beta (TGF-β) pathway. Thus, we aimed to investigate polymorphisms (rs1805110 and rs7526590) of the transforming growth factor beta receptor III gene (TGFBR3) with regard to laboratory biomarkers and clinical manifestations in individuals with SCA. Hematological, biochemical, immunological, and genetic analyses were carried out, as well as serum endothelin-1 measurements. The minor allele (A) of the TGFBR3 rs1805110 polymorphism was associated with increased hemoglobin, hematocrit, reticulocyte counts, total cholesterol, low-density lipoprotein, uric acid, and endothelin levels, as well as decreased platelet distribution width (PDW) and the occurrence of bone alterations. The minor allele (T) of TGFBR3 rs7526590 was associated with increased red cell distribution width, PDW, alkaline phosphatase, aspartate aminotransferase, total and indirect bilirubin, and lactate dehydrogenase levels, as well as lower ferritin levels and the occurrence of leg ulcers. Our data suggest that the minor allele (A) of TGFBR3 rs1805110 is associated with inflammation and bone alterations, while the minor allele (T) of TGFBR3 rs7526590 is related to hemolysis and the occurrence of leg ulcers.

Sickle cell disease (SCD) consists of a group of hemoglobinopathies in which individuals present highly variable clinical manifestations. Sickle cell anemia (SCA) is the most severe form, while SC hemoglobinopathy (HbSC) is thought to be milder. Thus, we investigated the clinical manifestations and laboratory parameters by comparing each SCD genotype. We designed a cross-sectional study including 126 SCA individuals and 55 HbSC individuals in steady-state. Hematological, biochemical and inflammatory characterization was performed as well as investigation of previous history of clinical events. SCA patients exhibited most prominent anemia, hemolysis, leukocytosis and inflammation, whereas HbSC patients had increased lipid determinations. The main cause of hospitalization was pain crises on both genotypes. Vaso-occlusive events and pain crises were associated with hematological, inflammatory and anemia biomarkers on both groups. Cluster analysis reveals hematological, inflammatory, hemolytic, endothelial dysfunction and anemia biomarkers in HbSC disease as well as SCA. The results found herein corroborate with previous studies suggesting that SCA and HbSC, although may be similar from the genetic point of view, exhibit different clinical manifestations and laboratory alterations which are useful to monitor the clinical course of each genotype.

This study investigated the effects of hydroxyurea (HU) on hematological, biochemical and inflammatory parameters in children with sickle cell anemia (SCA) in association with βS haplotype and α-thalassemia. We included 22 children with SCA who were followed for an average of 14.5 months. Laboratory parameters were assessed by electronic methods, and molecular analysis was investigated by PCR-RFLP and allele-specific PCR. Results showed significant increases in hemoglobin, HbF, hematocrit, MCV, MCH, glucose, HDL-C and albumin levels, as well as significant decreases in MCHC and AST levels, WBC, neutrophils, eosinophils, lymphocytes and reticulocytes, in children during HU therapy. HbF levels were positively correlated with hemoglobin, hematocrit, MCV and total protein, yet negatively correlated with MCHC, RDW, AAT and AST during HU therapy (p

We investigated the nasopharynx and oropharynx microbiota in sickle cell disease (SCD) to identify the microorganisms, antibiotic sensitivity, prevalent serotypes, and association of with laboratorial markers. Oropharynx/nasopharynx secretions were investigated in 143 SCD children aging 6 months to 17 years. Pathogens were isolated using standard procedures, and laboratorial markers were performed by automated methods. Staphylococcus aureus (S. aureus) was isolated from nasopharynx and oropharynx of 64 and of 17 SCD children respectively. Streptococcus pneumoniae (S. pneumoniae) was isolated from the nasopharynx and oropharynx of eight SCD patients. Serotypes of S. pneumoniae were 19F, 23F, and 14. All isolates were susceptible to penicillin, and patients whose nasopharynx and oropharynx were colonized by S. pneumoniae had high concentrations of aspartate transaminase, alanine transaminase, and ferritin. S. pneumoniae isolated were not penicillin-resistant serotypes suggesting that the use of penicillin for prophylaxis and/or treatment of infections is safe. Our finding of colonization and laboratory evaluation in SCD patients suggests that microorganisms are involved in the modulation of chronic inflammatory. The association of colonized microorganisms and laboratorial markers suggest a new approach to these patients follow-up, and additional studies of microorganism colonization and their association with SCD patients' clinical outcome will improve control and prevention strategies.

Background The search for sickle cell disease (SCD) prognosis biomarkers is a challenge. These markers identification can help to establish further therapy, later severe clinical complications and with patients follow-up. We attempted to study a possible involvement of levels of high-density lipoprotein cholesterol (HDL-C) in steady-state children with SCD, once that this lipid marker has been correlated with anti-inflammatory, anti-oxidative, anti-aggregation, anti-coagulant and pro-fibrinolytic activities, important aspects to be considered in sickle cell disease pathogenesis. Methods We prospectively analyzed biochemical, inflammatory and hematological biomarkers of 152 steady-state infants with SCD and 132 healthy subjects using immunochemistry, immunoassay and electronic cell counter respectively. Clinical data were collected from patient medical records. Results Of the 152 infants investigated had a significant positive association of high-density lipoprotein cholesterol with hemoglobin (P < 0.001), hematocrit (P < 0.001) and total cholesterol (P < 0.001) and a negative significant association with reticulocytes (P = 0.046), leukocytes (P = 0.015), monocytes (P = 0.004) and platelets (P = 0.005), bilirubins [total bilirubin (P < 0.001), direct bilirubin (P < 0.001) and indirect bilirubin (P < 0.001], iron (P < 0.001), aminotransferases [aspartate aminotransferase (P = 0.004), alanine aminotransferase (P = 0.035)], lactate dehydrogenase (P < 0.001), urea (P = 0.030), alpha 1-antitrypsin (P < 0.001), very low-density lipoprotein cholesterol (P = 0.003), triglycerides (P = 0.005) and hemoglobin S (P = 0.002). Low high-density lipoprotein cholesterol concentration was associated with the history of cardiac abnormalities (P = 0.025), pneumonia (P = 0.033) and blood transfusion use (P = 0.025). Lipids and inflammatory markers were associated with the presence of cholelithiasis. Conclusions We hypothesize that some SCD patients can have a specific dyslipidemic subphenotype characterized by low HDL-C with hypertriglyceridemia and high VLDL-C in association with other biomarkers, including those related to inflammation. This represents an important step toward a more reliable clinical prognosis. Additional studies are warranted to test this hypothesis and the probably mechanisms involved in this complex network of markers and their role in SCD pathogenesis.

Background: Sickle cell disease (SCD) is a group of inherited anemia characterized by heterogeneous clinical outcome, including hemolysis, chronic inflammation, and vaso-occlusive/painful crisis. Aims: We evaluated inflammatory and anti-inflammatory mediators, such as TNFα, IL-10, IL-12, IL-1β, IL-6, and IL-8, TGF-beta, tissue inhibitor of metalloproteinase (TIMP1), matrix metalloproteinase 9 (MMP9), heme, and leukotriene B4 (LTB4), and prostaglandin E2 (PGE2), the last two are products of the eicosanoid synthesis pathways, in SCD patients (in steady-state and in crisis-state) and healthy controls. For the same groups, in order to establish biomarkers of crisis and steady-state, we also investigated association among inflammatory/anti-inflammatory mediators and markers of hemolysis, inflammation, hepatic dysfunction, renal and lipid metabolism. Methods. We assessed 129 SCD steady-state patients (SP), 23 SCD in crisis patients (CP), and 67 healthy individuals (HC) age- and sex-matched with patients groups. Hematological analyzes were performed by automatic cell counter and hemoglobin profile by HPLC. Biochemistry analyses of inflammation and infection markers, as well as lipid, hepatic, and kidney metabolism markers were investigated by immunochemistry assays. Plasma levels of TNFα, IL-10, IL-12, IL-1β, IL-6 and IL-8 were measured using Cytometric Bead Array (CBA) according to the manufacturer's protocol. Plasma concentration of total heme was measured by QuantiChrom Heme Assay Kit. PGE2, LTB4, TGF-beta, TIMP1 and MMP9 levels were estimated in plasma samples and supernatants by ELISA, according to the manufacturer's instructions. Results . Steady-state SCD patients had the highest values of LTB4, PGE2, TIMP1, MMP9, IL-8, and IL-12 concentration compared with CP and HC groups (p < 0.0001). However, crisis-state SCD patients had the highest values of IL-1β, IL-6, IL-10, TNFα compared to HC and SP groups, and had a the lowest levels of LTB4, PGE2,TGF-β, MMP9, IL-8 and IL-12 levels (p < 0.0001 for both analysis). Significant difference of heme levels was not finding among the three groups. The ROC curve showed that LTB4, PGE2 and TGF- β are important markers related to steady state and IL-1β, IL-6, and TNFα are markers of crisis in SCD. Conclusions: The finding of difference in inflammatory markers level in steady state and crisis SCD patients suggest that these markers can be used to monitoring patients and to predict crisis events. The finding of similar heme concentration between steady-state and crisis-state SCD patients may be explained by the hyperhemolysis phenomenon, showing that even in steady-state, these patients continuous to have hemolysis and, consequently, continuous to generate heme and reactive oxygen species. Disclosures No relevant conflicts of interest to declare.

Sickle cell disease (SCD) has heterogeneous clinical picture and there are several pathway involved in SCD pathogenesis, and chronic inflammation and hemolysis are important hallmarks of the disease. Based on such points, there is a search for prognostic markers to establish possible sub-genotypes of the disease. The present study investigated the alpha-1 antitrypsin (AAT) levels, a serum glycoprotein inhibitor of proteases responsible for trigger inflammatory reactions, describing its associations with SERPINA1 gene polymorphisms, hematological and chemistry biomarkers, and clinical history in a group of SCD children in steady-state. The study was approved at FIOCRUZ research board and followed the principle of Declaration of Helsinki. A total of 356 steady state unrelated SCD patients were included at the present study and a control group (CG) compound by 100 unrelated healthy individuals sex and age matched with the patients group, which were from the same geographical origin. Patients age ranging 13.96+9.91 years of age, the AAT levels higher than the 50th percentile (158.0 mg/mL) had significantly lower red blood cells (RBC) count (p=0.003), hemoglobin (Hb) (p=0.0002) and hematocrit (Hct) (0.0002) concentration, and higher white blood cells (WBC) (p=0.004) and neutrophils (p=0.0001) counts, and higher C-reactive protein (CRP) levels (p Disclosures No relevant conflicts of interest to declare.

Background: Sickle cell disease (SCD) has a clinical heterogeneity and vasoocclusive events are an important clinical characteristic among patients. The paraoxonase 1 (PON 1) is a serum glycoprotein associated with the high-density lipoproteins cholesterol (HDL-C) and the variability of the PON1 activity has been associated with the PON1 genotypes. We tested a possible relevance of the PON1 in SCD patients in steady and crisis state, studying its activity and genotype, investigating M55L (rs854560) and R192Q (rs662) polymorphisms, and their association with markers of hemolysis, inflammation and organ dysfunction. Methods: The casuistic of the study was compound by 373 individuals, 154 SCD patients in steady state, 23 SCD patients in crisis, and 196 healthy controls. Chemistry, inflammatory and hematological biomarkers were investigated by chemiluminescence, immunoassay and electronic cell counter respectively. Molecular biology analyses were performed by PCR and PCR-RFLP techniques. Results: The allelic frequency of PON1192R among SCD patients group was 0.38 and of PON1192Q was 0.62. The allelic frequency of the control group individuals was 0.45 to the PON1192R and 0.55 to PON1192Q. The allelic frequency of PON1M55L gene polymorphism was 0.91 to the PON155L and 0.09 to the PON155M among the SCD patients group. The allelic frequency of the PON155L was 0.85 and for the PON155M was 0.15 among individuals of the control group. PON1 gene polymorphisms allelic frequencies were in Hardy-Weinberg equilibrium. Patients with genotype RR of PON1192 (p=0.0015) and MM of PON155 (p=0.0013) showed less PON1 activity than others genotypes. PON1 activity was higher among SCD patients in crisis and steady states than in control group individuals (p Conclusion: The PON1 activity and genetic alteration were not explored among SCD patients and may represent a new insight to explain endothelial dysfunction and oxidative stress reaction among these patients. Also, PON1 may represent an important molecule in SCD pathogenesis and the associated mechanisms need to be evaluated once it seems to be delivery from the HDL-C structure under a very intense hemolytic and inflammatory environment as happened in SCD, and may be explore as a further therapeutic strategy in disease. Disclosures No relevant conflicts of interest to declare.

Introduction Sickle cell disease (SCD) has a heterogeneous clinical picture, characterized by hemolysis, chronic inflammation and vaso-occlusive events and painful crisis. Aims Investigate the levels of Alpha 1 antitrypsin (A1AT), C reactive protein (CRP), hemolysis markers and its association with cytokines profile in SCD patients in steady-state and crisis-state. Methods It was developed a cross sectional study in a total of 72 SCD patients in steady-state (SP) and 23 SCD patients in crisis (CP) age-and-sex-matched. Hematological analysis was performed by automatic cell counter, hemoglobin profile by HPLC, and biochemistry analyses of inflammation and infection markers and lipid, hepatic, and kidney metabolism markers were investigated by immunochemistry assays. Plasma levels of TNFα, IFNγ, IL-10, IL-1β, IL-6 and IL-8 were measured using Cytometric Bead Array - CBA (BD Biosciences Pharmingen, USA) according to the manufacturer’s protocol. Results Statistical analysis showed that SCD steady-state patients group had the highest IL-12 concentration (p Conclusions Immunological aspects of SCD patients have been increasingly studied. The high levels of Th2 cytokines (IL-6 and IL-10) and proinflammatory cytokines (TNFα and IL-1β) in SCD patients in crisis-state, possibly is related to severe clinical manifestations, despite the fundamental role of these cytokines in the pathophysiology is not yet completely clear. The correlation between these cytokines and biomarkers associated with important clinical manifestations in SCD, such as WBC, platelet count, A1AT and CRP, highlights the important role of inflammatory mediators in SCD and its possible association with an expression or activation of adhesion molecules in neutrophils and vascular endothelium. Disclosures: No relevant conflicts of interest to declare.

Abstract 2140 Introduction: Nasopharynx and oropharynx bacterium colonization can be considered as initiation factor for local or systemic disease. The Streptococcus pneumoniae is a pathogen with epidemiological worldwide importance and it has been frequently associated with infection among SCD patients. The Staphylococcus aureus may also colonize the nasopharynx and has been associated with cause of infections in skin and soft tissue, pneumonia, sepsis, osteoarticulation. This study aimed to establish a profile of inflammation and hemolysis biomarkers of SCD individuals in association with the oropharyngeal and nasopharyngeal bacteria colonization. Patients and Methods: Biomarkers related to hemolysis and inflammation, including lipids and liver function, were determined by biochemical colorimetric reaction and also myeloperoxidase (MPO) gene polymorphisms were investigated with PCR and RFLP techniques. Medical history was obtained by patients' record. Thus, it was developed a cross-sectional study composed by 154 SCD children in a steady-state from the Bahia state, in Brazil, 68.2% (105/154) with HbSS genotype, and 31.8% (49/154) HbSC, in attendance of the outpatients clinic of the Foundation of Hematology and Hemotherapy of Bahia (HEMOBA). The study was approved by the Human research board from FIOCRUZ-BA and every official responsible signed an informed consent. Results: Nasopharynx and oropharynx colonization by S. pneumoniae was found in 14 (9.6%) SCD patients and by S. aureus in 81 (56.6%) patients, and both SCD genotypes had similar frequencies of these studied bacteria. It was not observed an increase in pneumococcal penicillin resistance. SCD patients with nasopharynx colonization by S. pneumoniae and S. aureus exhibited higher ferritin serum levels than patients with a normal microbiote respectively (p= Disclosures: No relevant conflicts of interest to declare.

Abstract 4811 Sickle cell disease (SCD) is a hematological disorder characterized by the S hemoglobin homozygosis or the association with others variants of hemoglobin. The SCD is characterized by heterogeneous clinical outcome compound by hemolysis, chronic inflammation and vaso-oclusive and pain episodes. Red Blood cell (RBC) count, hemoglobin concentration (HB), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), total bilirrubin (BT) and indirect bilirrubin (BI) are important biomarkers in SCD patients follow-up. The insulin are essential for regulation and maintenance of glucose homeostasis and the insulin resistance are associated with chronic sub-clinical inflammation and inflammatory markers such as C - reactive protein (CRP) and white blood cells (WBC) count. These markers have been correlated with the severity of SCD. The aim of this study was investigate insulin levels and HOMA-IR in SCD and it correlation with important biomarkers. Here we developed a cross sectional study involving 40 SCD patients in the steady-state of disease aging 13.9+ 8.23 years old. The RBC and WBC count were carried out by automated method and the levels of BT, BI, AST, LDH and insulin levels were investigated by immunochemistry assay method. The mean of insulin levels was 3.2 ± 2.2 and HOMA-IR was 0.61 ± 0.46. SCD patients with insulin levels lowest than the mean had an increase levels of AST (p Disclosures: No relevant conflicts of interest to declare.

Abstract 1547 Poster Board I-570 Introduction Levels of high-density lipoprotein cholesterol have been correlated with anti-inflammatory, anti-oxidative, anti-aggregation, anti-coagulant and pro-fibrinolytic activities. We hypothesized that lipoprotein cholesterol and triglycerides have important roles in sickle cell disease pathogenesis. Patients and Methods A prospective study of biochemical and hematological analyses of 152 steady-state children with sickle cell disease and 132 healthy subjects using immunochemistry, immunoassay and electronic cell counter respectively. Clinical data were collected from patient medical records. Data analyses were performed using Prism 5.01 (Graphpad Software, San Diego, CA), EPIinfo 6.04 (CDC, Atlanta, Georgia) and STATA SE 10 software (StataCorp, Texas, USA). Results There was a significant positive association of high-density lipoprotein cholesterol with hemoglobin (p Conclusion We hypothesize that some SCD patients can have a specific dyslipidemic subphenotype characterized by hypertriglyceridemia, high VLDL-C and low plasma LDL-C and HDL-C in association with other biomarkers, including those related to inflammation. This represents an important step toward a more reliable clinical prognosis. We suggest further studies and continued research into new mechanisms involving this complex network of markers in order to establish their role in SCD pathogenesis. Disclosures No relevant conflicts of interest to declare.