Elevated levels of autoantibodies against DNAJC2 in sera of patients with atherosclerotic diseases

Abstract

Background Serum antibody markers have been increasingly identified not only for cancer and autoimmune diseases but also for atherosclerosis-related diseases such as acute ischemic stroke (AIS), acute myocardial infarction (AMI), diabetes mellitus (DM), and chronic kidney disease (CKD). Biomarkers for transient ischemic attack (TIA) and non-ST segment elevation acute coronary syndrome (NSTEACS) are potentially useful for detection of early phase of atherosclerotic changes against AIS and AMI, respectively. Methods We utilized serological identification of antigens by recombinant cDNA expression cloning (SEREX) using a human aortic endothelial cell cDNA phage library and sera from patients with TIA or NSTEACS. Serum antibody levels were measured by amplified luminescent proximity homogeneous assay-linked immunosorbent assay (AlphaLISA) using purified recombinant antigens. Results Screening of sera from patients with TIA identified DnaJ heat shock protein family (Hsp40) member C2 (DNAJC2) as a candidate antigen, which was also isolated by SEREX screening using sera of patients with NSTEACS. The validation cohort revealed significantly higher DNAJC2 antibody (DNAJC2-Ab) levels in the sera of patients with TIA or AIS than those in healthy donors (HDs). Multivariate logistic regression analysis indicated that the predictive odds ratios (OR) of DNAJC2-Ab levels for TIA and AIS were 2.54 (95% confidence interval [CI]: 1.36–4.74, p = 0.0034) and 2.14 (95% CI: 1.39–3.30, p = 0.0005), respectively. Serum DNAJC2-Ab levels were also higher in patients with AMI, DM, and CKD than those in HDs. Conclusion Serum DNAJC2-Ab level may be useful for early detection of atherosclerotic lesions, which lead to AIS and AMI.

Anti-KIAA0513 Antibody as a Broad-spectrum Biomarker of Mortal Atherosclerotic and Cancerous Diseases

“…We incubated the reaction mixture at room temperature for 6-8 h, then added anti-human IgG-conjugated acceptor beads (2.5 µL at 40 µg/mL) and glutathione-conjugated donor beads (2.5 µL at 40 µg/mL), and incubated the mixture at room temperature in the dark for 1-14 days. We measured the chemical emissions using an EnSpire Alpha microplate reader (PerkinElmer), as described previously [5][6][7][8][9][10][11][12]. We calculated the speci c reactions by subtracting the emission counts of the GST control from the counts of GST-fusion proteins.…”

“…The membranes were washed 5 times with TBS-T and treated with HRP-conjugated secondary antibodies for 20 min. After 5 washes with TBS-T, the addition of Immobilon (Merck KGaA, Darmstadt, Germany) produced luminescence, which was detected by LuminoGraph II (Atto, Tokyo, Japan), as described previously [5,6,11]. Ampli ed luminescence proximity homogeneous assaylinked immunosorbent assay (AlphaLISA)…”

“…In terms of biomarker species, studies have reported enzyme, antigen, and, in recent years, nucleotide markers. However, there are still few reports on antibody markers, which include HSP60 [1], RPA2 [2], SOSTDC1 [3], PDCD11 [4], MMP1, CBX1, CBX5 [5], and DNAJC2 [6] for acute ischemic stroke (AIS); ATP2B4 [7], BMP-1 [2,7], DHPS [8], SH3BP5 [9], prolyl carboxypeptidase [10], LRPAP1 [11], and ASXL2 [12] for atherosclerosis; nardilysin/NRD1 [13] for acute cardiac syndrome; and TUBB2C [14], insulin [15], glutamic acid decarboxylase [16], adiponectin [17], and GADD34 [18] for diabetes mellitus (DM).…”

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

“…Previously, we searched for antibody markers using serological identification of antigens by cDNA expression cloning (SEREX) and the protein array method, and we reported on autoantibodies against Trop2/TACSTD2 [ 25 ], TRIM21 [ 26 ], Makorin 1 [ 27 ], and ECSA [ 28 ], for esophageal squamous cell carcinoma; FIR/PUF60 for colon cancer [ 29 ]; SH3GL1 [ 30 ] and filamin C [ 31 ] for glioma; EP300-interacting inhibitor of differentiation 3 for nonfunctional pancreatic neuroendocrine tumors [ 32 ]; proline-rich 13 for ulcerative colitis [ 33 ]; talin-1 for multiple sclerosis [ 34 ]; PSMA7 for amyotrophic lateral sclerosis [ 35 ]; NBL1/DAN [ 36 ] and SNX16 [ 37 ] for obstructive sleep apnea (OSA); and EXD2 for chronic thromboembolic pulmonary hypertension (CTEPH) [ 38 ]. We also reported on autoantibody markers for atherosclerosis-related diseases, e.g., RPA2 [ 39 ], PDCD11 [ 40 ], MMP1 [ 41 ], and DNAJC2 [ 42 ] for AIS; ASXL2 [ 43 ] for atherosclerosis; and nardilysin for acute coronary syndrome [ 44 ]. Here, we report on antibodies against death-inducer obliterator 1 (DIDO1), forkhead box J2 (FOXJ2), and cleavage and polyadenylation specificity factor (CPSF2) peptides, which are highly associated with AIS and could be useful as predictive markers.…”

Serum Anti-AP3D1 Antibodies Are Risk Factors for Acute Ischemic Stroke Related With Atherosclerosis

“…In this study, we were able to nd that the serum antibody levels against some of the SEREX antigens were elevated in patients compared with healthy donors (HDs) and reported TROP2, SLC2A1, TRIM21, and myomegalin as antibody biomarkers for esophageal squamous cell carcinoma (ESCC) [9][10][11][12] . SEREX was also introduced in screening the biomarkers for atherosclerosis and identi ed antibody biomarkers such as RPA2 9 , TUBB2C 13 , SH3BP5 14 , DHPS 15 , PDCD11 16,17 , MMP1, CBX1, CBX5 18 , DNAJC2 19 , ASXL2 20 , and LRPAP1 21 for atherosclerotic diseases including AIS and CVD. Notably, the antibody levels against DHPS, ATP2B4, BMP-1, ASXL2, and LRPAP1 were also elevated in patients with ESCC, which suggests the presence of multiple common biomarkers for atherosclerosis and cancer.…”