title: Development of immune cellular biosensing system for assessing chemicals on inducible nitric oxide synthase signaling activator.
abstract: An immune cellular biosensing system has been constructed to assess immunomodulating effects of chemicals. Production of nitric oxide in the immune cellular biosensing system was used as readout of an immune cellular response for assessing the immunomodulating effects of chemicals. The macrophage-like cell line RAW264.7, which has signaling pathways of inducible nitric oxide synthase, was employed in the cellular biosensing system. The immune cellular biosensing system consisted of a Pt counter electrode, an Ag/AgCl reference electrode, and a gold electrode onto which a polyion complex layer was coated to allow adherence of the RAW264.7 cells. As the results of evaluating effects of a polyion complex layer on cell viabilities by using WST-8 assay, the polyion complex layer did not affect RAW264.7 cells. The polyion-coated gold electrode could measure NO without the drawback of electrochemical interference that occurs with differential pulse voltammetry. The detection limit of the immune cellular biosensing system was 4.2 nM released NO as measured by double potential step chronoamperometry. The potent immune activating abilities of lipopolysaccharide and interferon-gamma could be assessed by the cellular biosensing system; NO release from cells was detected within 600 ms.
authors.affiliation: Department of Biological Information, Tokyo Institute of Technology, 4259, Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
authors.forename: Ken-ichiro
authors.initials: K
authors.lastname: Kamei
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Haruyama
authors.forename: Masayasu
authors.initials: M
authors.lastname: Mie
authors.forename: Yasuko
authors.initials: Y
authors.lastname: Yanagida
authors.forename: Masuo
authors.initials: M
authors.lastname: Aizawa
authors.forename: Eiry
authors.initials: E
authors.lastname: Kobatake
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology*; D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D002470:Cell Survival; D003124:Colorimetry / Q000379:methods; D004566:Electrodes; D006046:Gold / Q000737:chemistry; D007371:Interferon-gamma / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000737:chemistry / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D009569:Nitric Oxide / Q000032:analysis* / Q000737:chemistry; D019001:Nitric Oxide Synthase / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II; D011107:Polylysine / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D015398:Signal Transduction; D013451:Sulfonic Acids / Q000737:chemistry
chemical_list: D000276:Adjuvants, Immunologic; D008070:Lipopolysaccharides; D011108:Polymers; D013451:Sulfonic Acids; D011107:Polylysine; D009569:Nitric Oxide; D006046:Gold; D007371:Interferon-gamma; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; C077114:styrenesulfonic acid polymer