PB1399 - NETosis in Sickle Cell Disease

Background: Inflammation, thrombosis are increased in sickle cell anemia. NETosis is increased in inflammatory and malignant diseases and is associated with thrombosis.

Aims: Aim was to demonstrate the increased inflammation in sickle cell anemia by demonstrating the activity of NETosis and to regulate the prophylaxis necessary for life-threatening thrombosis, which may occur at the end of this process.

Methods: Blood samples of patients and healthy persons were taken. Twenty-five patients and 25 healthy individuals were included in the study. 2 mL venous blood samples of cases, EDTA tube for neutrophil elastase enzyme level, received.. Blood samples taken into EDTA tube for neutrophil isolation were centrifuged at 4000 rpm for 10 minutes and stored at +4 oC.. All blood samples were studied simultaneously. Neutrophils obtained using MACSxpress® Neutrophil Isolation Kit Human were used for NETs formation. In the NETosis ASSAY Kit stage, cells were suspended with NET assay buffer and incubated, then centrifuged to collect neutrophil elastase enzyme activity.The supernatants were incubated with the addition of NET Assay Neutrophil Elastase Substrate and the film on the plate was opened at 405 nm. The concentration absorbance graph was drawn by utilizing the standards and the calculations were calculated. Blood samples from patient and control groups were prepared for appropriate fluorescence and scanning electron microscope images and prepared with appropriate protocols.

Results: Although no statistically significant difference was found, we showed that NETosis formation increases in patients with sickle cell anemia compared to healthy subjects (p=0,329). We also showed that NETosis formation increses with the increasing number of painfull crisis in these patients.

Conclusion(s): It has been shown that NETosis formation increases in patients with sickle cell anemia compared to individuals with health subjects. However, more studies with larger number of cases are needed for understnading NETosis in the ethiopathogenesis of sickle cell disease.