Yoshida Yoichi, Zhang Xiao-Meng, Wang Hao, Machida Toshio, Mine Seiichiro, Kobayashi Eiichi, Adachi Akihiko, Matsutani Tomoo, Kamitsukasa Ikuo, Wada Takeshi, Aotsuka Akiyo, Iwase Katsuro, Tomiyoshi Go, Nakamura Rika, Shinmen Natsuko, Kuroda Hideyuki, Takizawa Hirotaka, Kashiwado Koichi, Shin Hideo, Akaogi Yuichi, Shimada Junichiro, Nishi Eiichiro, Ohno Mikiko, Takemoto Minoru, Yokote Koutaro, Kitamura Kenichiro, Iwadate Yasuo, Hiwasa Takaki
Department of Neurological Surgery, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan.
Department of Biochemistry and Genetics, Graduate School of Medicine, Chiba University, Chiba 260-8670, Japan.
Heliyon. 2020 Aug 19;6(8):e04661. doi: 10.1016/j.heliyon.2020.e04661. eCollection 2020 Aug.
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.
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.
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, = 0.0034) and 2.14 (95% CI: 1.39-3.30, = 0.0005), respectively. Serum DNAJC2-Ab levels were also higher in patients with AMI, DM, and CKD than those in HDs.
Serum DNAJC2-Ab level may be useful for early detection of atherosclerotic lesions, which lead to AIS and AMI.
血清抗体标志物不仅越来越多地被发现可用于癌症和自身免疫性疾病，还可用于动脉粥样硬化相关疾病，如急性缺血性卒中（AIS）、急性心肌梗死（AMI）、糖尿病（DM）和慢性肾脏病（CKD）。短暂性脑缺血发作（TIA）和非ST段抬高型急性冠状动脉综合征（NSTEACS）的生物标志物分别可能有助于检测针对AIS和AMI的动脉粥样硬化改变的早期阶段。
我们利用重组cDNA表达克隆（SEREX）技术，使用人主动脉内皮细胞cDNA噬菌体文库以及TIA或NSTEACS患者的血清进行抗原的血清学鉴定。使用纯化的重组抗原，通过放大发光邻近均相分析-酶联免疫吸附测定（AlphaLISA）测量血清抗体水平。
对TIA患者血清的筛选确定DnaJ热休克蛋白家族（Hsp40）成员C2（DNAJC2）为候选抗原，使用NSTEACS患者的血清进行SEREX筛选也分离出该抗原。验证队列显示，TIA或AIS患者血清中的DNAJC2抗体（DNAJC2-Ab）水平显著高于健康供体（HDs）。多因素逻辑回归分析表明，DNAJC2-Ab水平对TIA和AIS的预测比值比（OR）分别为2.54（95%置信区间[CI]：1.36-4.74，P = 0.0034）和2.14（95%CI：1.39-3.30，P = 0.0005）。AMI、DM和CKD患者的血清DNAJC2-Ab水平也高于HDs。
血清DNAJC2-Ab水平可能有助于早期检测导致AIS和AMI的动脉粥样硬化病变。
