HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE ADVANCED SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Kuribayashi, F. Articles by Nakamura, M. Search for Related Content PubMed Articles by Kuribayashi, F. Articles by Nakamura, M.

¹ Department of Host-Defense Biochemistry, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan
² Division of Pediatrics, Shizuoka City Hospital, 10-93 Ohtemachi, Aoi-ku, Shizuoka 420-8630, Japan
³ Department of Information and Communication Technology, School of High-Technology for Human Welfare, Tokai University, 317 Nishino, Numazu, Shizuoka 410-0395, Japan
⁴ Department of Pediatrics, Faculty of Medicine, Miyazaki University, 5200 Kihara, Kiyotake, Miyazaki 889-1692, Japan
⁵ Department of Basic Medical Sciences, Institute of Medical Science, University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108-8639, Japan
⁶ Central Lab, Effector Cell Institute, Inc., 4-7-7 Aobadai, Meguro-ku, Tokyo 153-0042, Japan

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 ~45x 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.

^* Correspondence: kurikuri{at}nagasaki-u.ac.jp