112 Laboratory, School of Chemistry and Biological Engineering, University of Science and Technology Beijing, Beijing, China.
Department of Pediatrics, Beijing Children's Hospital Affiliated to Capital Medical University, Beijing, China.
Clin Exp Immunol. 2018 Dec;194(3):371-379. doi: 10.1111/cei.13204. Epub 2018 Oct 15.
Kawasaki disease (KD) is an immune-mediated vasculitis with symptoms that mimic febrile illness; the immune origin has been suggested but never proved. First, cell chip technology was used to screen immune targets cells. With the indirect immunofluorescence assay we found that the HeLa cell chip could be recognized by KD patient serum samples. The target cell proteome was extracted as antigens, and antigen recognition reaction was performed using the patients' serum as antibodies and the detected target protein was detected and identified as phosphoglycerate kinase 1 (PGK1). Then PGK1 was produced and tested with enzyme-linked immunosorbent assay (ELISA), Western blotting, immunoprecipitation and competitive inhibition immunofluorescence assay. Immunoglobulin (Ig)G against PGK1 was detected in 46% (23 of 50) sera of KD patients, 13% (five of 38) sera in febrile non-KD patients (FC) and 2·6% (one of 38) sera in healthy donors. As an immune target, PGK1 not only helps understanding of the pathogenesis, it also has potential value in facilitating the laboratory diagnosis of KD.
川崎病(KD)是一种免疫介导的血管炎,其症状类似于发热性疾病;免疫起源已被提出,但从未得到证实。首先,我们使用细胞芯片技术筛选免疫靶细胞。通过间接免疫荧光法,我们发现 HeLa 细胞芯片可被 KD 患者血清样本识别。提取靶细胞蛋白质组作为抗原,以患者血清作为抗体进行抗原识别反应,检测到的靶蛋白被鉴定为磷酸甘油酸激酶 1(PGK1)。然后通过酶联免疫吸附试验(ELISA)、Western blot、免疫沉淀和竞争抑制免疫荧光法来检测和验证 PGK1。在 50 例 KD 患者的 46%(23 例)血清中检测到针对 PGK1 的 IgG,38 例发热非 KD 患者(FC)血清中为 13%(5 例),38 例健康供体血清中为 2.6%(1 例)。PGK1 作为一种免疫靶标,不仅有助于了解发病机制,而且在促进 KD 的实验室诊断方面具有潜在价值。