Serum anti-DIDO1, anti-CPSF2, and anti-FOXJ2 antibodies as predictive risk markers for acute ischemic stroke

Abstract

Background Acute ischemic stroke (AIS) is a serious cause of mortality and disability. AIS is a serious cause of mortality and disability. Early diagnosis of atherosclerosis, which is the major cause of AIS, allows therapeutic intervention before the onset, leading to prevention of AIS. Methods Serological identification by cDNA expression cDNA libraries and the protein array method were used for the screening of antigens recognized by serum IgG antibodies in patients with atherosclerosis. Recombinant proteins or synthetic peptides derived from candidate antigens were used as antigens to compare serum IgG levels between healthy donors (HDs) and patients with atherosclerosis-related disease using the amplified luminescent proximity homogeneous assay-linked immunosorbent assay. Results The first screening using the protein array method identified death-inducer obliterator 1 (DIDO1), forkhead box J2 (FOXJ2), and cleavage and polyadenylation specificity factor (CPSF2) as the target antigens of serum IgG antibodies in patients with AIS. Then, we prepared various antigens including glutathione S-transferase-fused DIDO1 protein as well as peptides of the amino acids 297–311 of DIDO1, 426–440 of FOXJ2, and 607–621 of CPSF2 to examine serum antibody levels. Compared with HDs, a significant increase in antibody levels of the DIDO1 protein and peptide in patients with AIS, transient ischemic attack (TIA), and chronic kidney disease (CKD) but not in those with acute myocardial infarction and diabetes mellitus (DM). Serum anti-FOXJ2 antibody levels were elevated in most patients with atherosclerosis-related diseases, whereas serum anti-CPSF2 antibody levels were associated with AIS, TIA, and DM. Receiver operating characteristic curves showed that serum DIDO1 antibody levels were highly associated with CKD, and correlation analysis revealed that serum anti-FOXJ2 antibody levels were associated with hypertension. A prospective case–control study on ischemic stroke verified that the serum antibody levels of the DIDO1 protein and DIDO1, FOXJ2, and CPSF2 peptides showed significantly higher odds ratios with a risk of AIS in patients with the highest quartile than in those with the lowest quartile, indicating that these antibody markers are useful as risk factors for AIS. Conclusions Serum antibody levels of DIDO1, FOXJ2, and CPSF2 are useful in predicting the onset of atherosclerosis-related AIS caused by kidney failure, hypertension, and DM, respectively.

Serum Anti-BRAT1 is a Common Molecular Biomarker for Gastrointestinal Cancers and Atherosclerosis

“…An improved version of the aforementioned method was used to screen clones that were immunoreactive to the sera of patients with TIA ( 31 ). A human aortic endothelial cell cDNA expression library (Uni-ZAP XR Premade Library, Stratagene, La Jolla, CA, USA) was transfected into Escherichia coli ( E. coli ) XL1−Blue MRF′ (Stratagene).…”

“…SEREX screens antigens related to tumors and autoimmune diseases including GI cancers [EC ( 21 , 22 ), GC ( 23 ), LC ( 24 ), and CRC ( 25 ) and PC ( 26 )], multiple sclerosis (MS) ( 27 ), systemic sclerosis (SSc) ( 28 ), systemic lupus erythematosus (SLE) ( 29 ), rheumatoid arthritis (RA) ( 30 ), and others. In earlier studies, we successfully applied SEREX to AS-related diseases and identified antibodies against CPSF2, DIDO1, FOXJ2, MMP1, CBX1, CBX5, and LAMP1 in TIA and CI ( 31 – 33 ). We also identified antibodies against TUBB2C in the sera of DM patients ( 34 ).…”

Serum anti-SERPINE1 antibody as a potential biomarker of acute cerebral infarction

“…The mixture was incubated further for 7–21 days at room temperature in the dark. On days 7, 14, and 21, chemiluminescence was read on an EnSpire Alpha microplate reader (Perkin Elmer) as previously described 12 , 20 , 21 , 27 , 28 , 31 – 34 . The results measured on day 14, when the specific response was highest, were used in the analysis.…”

GADD34 activates p53 and may have utility as a marker of atherosclerosis

“…Sera were collected from patients after provision of written informed consent. Serum samples from healthy donors (HD) and patients with AIS or transient ischemic attack (TIA) were obtained from Sawara Hospital as described previously [15]. HD had no abnormalities on cranial magnetic resonance imaging.…”

“…AlphaLISA (ampli ed luminescence proximity homogeneous assay-linked immunosorbent assay) Antigenic GST-fused GADD34 was expressed in Escherichia coli and puri ed as described previously [12,15]. AlphaLISA was used to measure serum antibody levels.…”