High-Throughput Chip Assay for Investigating Escherichia coli Interaction with the Blood–Brain Barrier Using Microbial and Human Proteome Microarrays (Dual-Microarray Technology)

Bacterial meningitis in neonates and infants is an acute lethal disease and occurs in response to microbial exploitation of the blood–brain barrier (BBB), resulting in the intracranial inflammation. Several pathogens, such as Escherichia coli (E. coli), can cause this devastating disease; however, the underlying molecular mechanisms by which these pathogens exploit the BBB remain incompletely understood. To identify important players on both the pathogen and host sides that govern the E. coli–BBB cell interactions, we took advantage of the E. coli and human proteome microarrays (i.e., HuProt) as an unbiased, proteome-wide tool for identification of important players on both sides. Using the E. coli proteome microarrays, we developed a unique high throughput chip-based cell probing assay to probe with fluorescent live human brain microvascular endothelial cells (HBMEC, which constitute the BBB). We identified several transmembrane proteins, which effectively bound to live HBMEC. We focused on YojI protein ...