title: Chemiluminometric immuno-analysis of innate immune response against repetitive bacterial stimulations for the same mammalian cells.
abstract: For monitoring of human cellular response to repetitive bacterial stimulations (e.g., Pseudomonas aeruginosa in a lysate form), we devised a chemiluminescent immuno-analytical system for toll-like receptor 1 (TLR1) as marker present on cell surfaces (e.g., A549). Upon stimulation, TLR1 recognizes pathogen-associated molecular patterns of the infectious agent and are then up-regulated via activation of the nuclear factor-κB (NF-κB) pathway. In this study, the receptor density was quantified by employing an antibody specific to the target receptor and by producing a chemiluminometric signal from an enzyme labeled to the binder. The activated status was then switched back to normal down-regulated stage, by changing the culture medium to one containing animal serum. The major factors affecting activation were the stimulation dose of the bacterial lysate, stimulation timing during starvation, and up- and down-regulation time intervals. Reiterative TLR regulation switching up to three times was not affected by either antibody remained after immunoassay or enzyme substrate (e.g., hydrogen peroxide) in solution. This immuno-analysis for TLRs could be unique to acquire accumulated response of the human cells to repeated stimulations and, therefore, can eventually apply to persistency testing of the cellular regulation in screening of anti-inflammatory substances.
authors.affiliation: Department of Bio-Microsystem Technology, Korea University, 1, 5-ka, Anam-dong, Seongbuk-Gu, Seoul 136-701, Korea.
authors.forename: Jin-Woo
authors.initials: JW
authors.lastname: Jeon
authors.affiliation: Department of Bio-Microsystem Technology, Korea University, 1, 5-ka, Anam-dong, Seongbuk-Gu, Seoul 136-701, Korea.
authors.forename: Il-Hoon
authors.initials: IH
authors.lastname: Cho
authors.affiliation: Department of Biotechnology and Bioinformatics, Korea University, 2511 Sejong-ro, Sejong 339-700, Korea.
authors.forename: Un-Hwan
authors.initials: UH
authors.lastname: Ha
authors.affiliation: Department of Electronics &Information Engineering, Korea University, 2511 Sejong-ro, Sejong 339-700, Korea.
authors.forename: Sung-Kyu
authors.initials: SK
authors.lastname: Seo
authors.affiliation: 1] Department of Bio-Microsystem Technology, Korea University, 1, 5-ka, Anam-dong, Seongbuk-Gu, Seoul 136-701, Korea [2] Department of Biotechnology and Bioinformatics, Korea University, 2511 Sejong-ro, Sejong 339-700, Korea.
authors.forename: Se-Hwan
authors.initials: SH
authors.lastname: Paek
mesh_terms: D045744:Cell Line, Tumor; D006801:Humans; D007113:Immunity, Innate; D008163:Luminescent Measurements; D016328:NF-kappa B / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000472:pathogenicity*; D051194:Toll-Like Receptor 1 / Q000235:genetics / Q000378:metabolism; D051193:Toll-Like Receptors / Q000235:genetics / Q000378:metabolism*; D015854:Up-Regulation
chemical_list: D016328:NF-kappa B; D051194:Toll-Like Receptor 1; D051193:Toll-Like Receptors
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