川崎医科大学 教員情報   
     


  クリバヤシ フトシ   Futoshi Kuribayashi
  栗林 太
   所属   川崎医科大学  医学部 基礎医学 生化学
   職種   教授
論文種別 原著
単著/共著の別 共著
言語種別 英語
査読の有無 査読あり
表題 Cell adhesion markedly increases lucigenin-enhanced chemiluminescence of the phagocyte NADPH oxidase
掲載誌名 正式名:Genes to Cells
略  称:Genes to Cells
巻・号・頁 13(12),1249-1256頁
著者・共著者 Kuribayashi Futoshi, Tsuruta Satoru, Yamazaki Tsuyoshi, Nunoi Hiroyuki, Imajoh-Ohmi Shinobu, Kanegasaki Shiro, Nakamura Michio
発行年月 2008/11
概要 Lucigenin-enhanced chemiluminescence (LECL) is widely used for the detection of reactive oxygen species released from various cells and mitochondria. However, the LECL response varies depending on cell species and assay conditions at least in part by unknown factors. Here we report that cell adhesion is an important factor for increasing LECL of tetradecanoylphorbol acetate (TPA)-stimulated human neutrophils. More than 90% LECL remained even after complete removal of the cell suspension 10 min after TPA stimulation, and ~22.5% of neutrophils were adhered to the reaction tube. These results indicate that LECL by an adhering neutrophil is ~45× higher than that by a non-adhering neutrophil. LECL by leukocyte adhesion deficiency neutrophils was one-fifth of that by normal neutrophils and completely disappeared when the cell suspension was removed, confirming that LECL depends highly on cell adhesion. The oxidase activity of adhering neutrophils measured after permeabilization with Renex 30 together with NADPH addition was similar to that of non-adhering neutrophils, indicating that lucigenin and cell adhesion do not enhance the oxidase activity. Based on these findings, we propose that a mixture of adhering and non-adhering neutrophils can be used for simultaneous screenings of adhering activity and the oxidase activity of neutrophils.
DOI 10.1111/j.1365-2443.2008.01241