Abstract
Cerebral infarction (CI), cardiovascular disease (CVD), diabetes mellitus (DM) and chronic kidney disease (CKD) are atherosclerosis-related diseases, which are major causes of health damage. For early and sensitive diagnosis, development of novel biomarkers is expected and of significant practical importance. First screening was carried out by phage expression cloning to identify antigen proteins recognized by serum IgG antibodies in patients with atherosclerosis. RPA2, LRPAP1, EEF1A1, SPOCK1, LOC729260, tubulin beta 2C (TUBB2C) and KIAA0020 markers were identified. We then compared the serum antibody levels against the candidate proteins between healthy donors (HD) and patients with CI, CVD, DM, or CKD by Alpha (amplified luminescent proximity homogeneous assay)-LISA method. The results showed that the serum TUBB2C antibody levels were significantly higher in patients with CI, DM, or CKD than those in HD. Using the average + 2SD of HD as the cut-off value, the positive thresholds of TUBB2C antibody markers were 14.8% in CI, 25.8% in DM, and 18.3% in CKD. TUBB2C antibody levels were well correlated with artery stenosis degrees such as plaque score, maximum intima-media thickness and cardio ankle vascular index. Consequently, TUBB2C antibody markers are useful to diagnose atherosclerosis, DM, and CKD, and can be applied to the prediction of the onset of CI. The serum anti-TUBB2C antibody markers are useful for the diagnosis of DM and CKD. Keywords: Atherosclerosis; Diabetes mellitus; Chronic kidney disease; Cerebral infarction; Cardiovascular disease; Antibody biomarker
Highlights
Cerebrovascular disease occupies approximately 10% of the cause of mortality in Japan
N-terminal biotinylated 14mer peptides corresponding to the amino acid sequences of the positive spots of peptide arrays were synthesized and used for AlphaLISA
lipoproteinrelated protein-associated protein 1 (LRPAP1), eukaryotic translation elongation factor 1 alpha 1 (EEF1A1), LOC729260 and tubulin beta 2C (TUBB2C) proteins were expressed as GST-fusion proteins in E. coli and purified by affinity-chromatography using GSTSepharose as described previously [9,10,11,21]
Summary
Cerebrovascular disease occupies approximately 10% of the cause of mortality in Japan. After the onset of a stroke, patients have suffered from prognostic symptoms for a long time, and the disease is a major cause of being bedridden. For the diagnosis of CI, various risk factors have been developed including body mass index (BMI), low density lipoprotein (LDL) cholesterol, uric acid, smoking habits and family history [4,5,6]. Diabetes mellitus (DM) is a major risk factors of CI and cardiovascular diseases (CVD) [7]. Prevention of CI and CVD has been attempted by using DM markers, which are blood glucose, glycoalbumin and glycohemoglobin (HbA1c) [8]. Despite frequent application for diagnosis, prevention of the onset remains unsuccessful.
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