The peripheral blood of Aβ binding RBC as a biomarker for diagnosis of Alzheimer's disease

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Abstract
Background:in vitro, it has been reported that amyloid β (Aβ) is bound to red blood cells (RBCs) and this process damages the red cell. Also, a possible relationship between RBCs and Alzheimer's disease (AD) is supported by the findings of RBC impairment in AD. Therefore, Aβ fibrils bounding RBC are of great interest as potential biomarkers. Methods: in this study, we focused on Aβ amyloid fibrils and/or aggregation on the peripheral RBC from 50 subjects with AD and 50 healthy controls (HCs) through thioflavin T (ThT) staining followed by immunofluorescence assay to confirm the presence of Aβ amyloid fibrils and/or aggregation on the RBC. Then we optimised fluorescence staining and imaging conditions and analysed the images obtained by image processing software. Results: we have analysed RBC morphology in blood from 50 subjects with AD and 50 HCs found that 16.8% of the RBCs are elongated as compared with 6.7% in normal controls (P < 0.01), and there is a negative correlation between the two parameters (P < 0.05). Our study showed that 98% of AD peripheral RBCs were amyloid binding-positive (ranging from 2 to 30%), while only 38% that of RBCs (ranging from 2 to 3.4%) were in HCs. We also found four modified morphologies of RBCs triggered by Aβ binding, which may serve as an ancillary investigation and indicate the progression of AD. Conclusion: we first directly prove the existence of Aβ binding RBCs in peripheral blood. In addition, we observed new modified morphologies of RBC triggered by Aβ binding, all of those can serve as a biomarker for the diagnosis and progression of AD.

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