Immunoglobulin Analysis of Kawasaki Disease Using E. Coli Protome Microarrays

Kawasaki disease (KD) is a systemic vasculitis of unknown etiology that primarily affects children less than 5 years of age. The biological markers specific for the diagnosis of KD is still unavailable till now. We wish to find some diagnostic markers for clinician and clinical use. From our previous results, we found that both T helper (Th) 2 and Th1 immune reaction associated with CAL and intravenous immunoglobulin (IVIG) treatment response in KD. Genome wide association study (GWAS) reported immune genes including ITPKC, CD40, BLK and FCGR2A playing important role in the pathogenesis of KD and the formation of CAL (Nature Genetics 2011, 2012). Immunologic response showed more predominate than infectious agent in KD. E. coli, a kind of common intestine bacteria in humans, has been known as one of the first foreign substances that a newborn infant encounters. Proteome microarray has recently emerged as a powerful tool for high-throughput protein interaction studies, and has been applied in many kinds of systematic investigations in biological and biochemistry researches. In Dr. Chen Jason (from Cheng Kung University, Tainan, Taiwan) lab, they have the entire expression library of E. coli K12, which are about 4200 genes. (Nature Methods 2008) He used the library to purify almost all the 4,200 proteins and printed them on glass slides to form E. coli proteome chips. We cooperated with Prof. Chen and used these chips to analyze the E. coli proteome interactions with children serological immunoglobulin by incubating the proteome chips with serum from KD patients. In our preliminary data (that was published on Mol Cell Proteomics. 2018 and award of National Innovation Award), we found several sets of E. coli proteins which can classify into highly and lowly immunogenic groups by the recognition to children serological immunoglobulin. Thus, we hypothesize that certain immunoglobulin profile is responsible for the susceptibility and/or CAL formation in KD and had difference profile pattern in different stage of KD. Based on the hypothesis above, we attempt to reach the 3 specific aims in the following 3 years below: 1. In the first year, we will enroll more cases to compare the immunoglobulin profiles between Kawasaki disease children, febrile control and normal children even from area in China. Certain profile of immunoglobulin will help the differential diagnosis of KD in the early stage of fever. Our aim is to set up the protein array chip specific for KD diagnosis. 2. In the second year, we will compare the immunoglobulin profiles from different stage of KD (before IVIG, after IVIG, 1 month later, 1 year later and 2 year later) and analyze the association with coronary artery lesions and IVIG treatment response. Immune response of Th1/Th2/Th17 will also analyzed. 3. In the third year, we will further compare the immunoglobulin profiles from different age of the same patients including childhood, early youth, adolescent and youth (through call back the previous KD patients) and analyze different immunoglobulin profile. Gut microbiota will also enrolled for further analysis.

Project ID:PC10901-1752
External Project ID:MOST108-2314-B182-037-MY3