Measurement of chemiluminescence from neutrophils in a 96-well microplate using Lumi Box U-800 II

Kikuchi, Takashi; Suzuki, Katsuhiko; Abé, Tatsuya; Satoh, Hideki; Endoh, Tetsu; Hasegawa, Hiroko; Nakaji, Shigeyuki; Sugawara, Kazuo; Kumae, Takashi

doi:10.1002/(sici)1099-1271(199705/06)12:3<149::aid-bio440>3.0.co;2-o

Cited by 30 publications

(20 citation statements)

References 10 publications

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“…Opsonization of particles. Zymosan from Saccharomyces cerevisiae, which is a well-known activator of the alternative pathway of complement system (19,23), was employed for opsonized particles. Zymosan A (Sigma, USA) was suspended in Hanks' balanced salt solution (HBSS) at a concentration of 5 mg/mL and then opsonization was performed by adding the final concentration of 13% of serum samples and incubating at 37°C for 30 min.…”

Section: Measurement Of Soamentioning

confidence: 99%

Depressed humoral immunity after weight reduction in competitive judoists

et al. 2002

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We studied changes in serum opsonic activity (SOA) in male judoists who were engaged in active weight reduction. Serum immunoglobulins, complements and SOA, measured by neutrophil-associated chemiluminescence responses, were investigated 20 days, 7 days and 1 day before a competition and 5 days after the competition. In addition, muscle strength and anaerobic work capacity, as well as body composition, were also determined. A dietary survey was conducted daily during the observation period. Body weight decreased by 4.2 kg over 19 days. SOA significantly decreased 5 days after the competition, as well as the concentrations of serum immunoglobulins, complements and total proteins. These trends were noted in the marked weight reduction group (i.e. reduction weight of body fat/body fat weight before weight reduction > or = 25%) more than the slight reduction group (<25%). Depressed SOA was closely correlated with the decreased concentrations of immunoglobulins and complements. These results suggest that the decrease in immunoglobulins and complements following weight reduction is associated with reduced SOA, which might cause susceptibility to infections. This study demonstrated that such immunosuppression appeared in the recovery period after the competition rather than immediately before the competition.

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Section: Measurement Of Soamentioning

confidence: 99%

Depressed humoral immunity after weight reduction in competitive judoists

et al. 2002

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“…The application of charge-coupled devices (CCDs) in the bio-sciences has been reported as a high-sensitivity technique for measuring luminescence (9,10). We have previously reported this measurement technique for chemiluminescence, using a parallel luminometer with a cooled CCD camera (11), and a new software was developed to decrease the background noise for more sensitive measurement (12). The parallel luminometer can measure 96 samples simultaneously with excellent reproducibility when a 96-well microplate is used, and we use a standardized light emission microplate (Luminometer Check Plate, Berthold Japan Co.) to control the inter-measurement variation.…”

Section: Introductionmentioning

confidence: 99%

Assessment of training effects on activity levels of alveolar macrophage in matured rats using chemiluminescent technique

Kumae¹,

Arakawa²

2003

Luminescence

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Chemiluminescence responses have been used for the evaluation of phagocyte function. In this study, to evaluate effects of training started after maturation on pulmonary immunity, the activity levels of rat alveolar macrophages (AMs) were assessed as reactive oxygen species (ROS) generating capacity, measured by lucigenin- and luminol-dependent chemiluminescence, using a parallel luminometer. One group of male Wistar rats started training at 11 weeks old and another group at 17 weeks old. The experimental period was 12 weeks, and about half of the rats were sacrificed after 6 weeks. The forced and voluntary exercises affect the mean levels of body weights and cell populations in the bronchoalveolar lavage fluid in younger animals; however, the voluntary exercise group in younger animals seemed to adapt after 12 weeks. By contrast, chemiluminescence responses in older rats observed after 6 weeks suggest that AMs are primed, and the maximum releasing activities of ROS are reduced. These changes in AM activity may be caused by the exercise and separation stresses and the rats may adapt to those stressors after 12 weeks. The chemiluminescent technique is thought to be useful to evaluate the changes of AM activity.

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“…Neutrophils were isolated from peripheral blood collected from the median cubital vein of 10 healthy male adults aged 21-28 (mean = 25) by a gradient centrifugation method (9,10). The neutrophil count was measured using a Sysmex F-300 automatic blood cell counter (TOA Medical Electronics, Kobe, Japan).…”

Section: Isolation Of Human Neutrophilsmentioning

confidence: 99%

“…The stimulants used were opsonized zymosan (OZ) and 12-myristate 13-acetate phorbol (PMA; Sigma, St. Louis, USA). OZ was prepared by opsonizing Zymosan A (Sigma) using pooled sera according to the previously described steps (9,10). OZ was suspended in HBSS at concentrations of 5 mg/mL for LgCL and 1 mg/mL for luminol-dependent LmCL before use (10).…”

Section: Neutrophil-stimulating Factorsmentioning

confidence: 99%

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Effect of interferon‐α on production of reactive oxygen species by human neutrophils

et al. 2001

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In order to examine that the effects of interferon alpha (IFN-alpha) on the production of reactive oxygen species from human neutrophils, we investigated in vitro effects of IFN-alpha on neutrophil chemiluminescence responses. Human neutrophils were incubated with 0-1 x 10(5) IU/mL IFN-alpha for 10-60 min just before stimulation with opsonized zymosan and phorbol 12-myristate 13-acetate. Luminol-dependent (LmCL) and lucigenin-dependent (LgCL) chemiluminescence responses were measured using a Lumi Box H-1000. The LmCL response was inhibited by pre-incubation with high concentrations of IFN-alpha (more than 1 x 10(4) IU/mL). In contrast, LgCL response was increased when neutrophils were pre-incubated with high concentration of IFN-alpha. Effects of pre-incubation of IFN-alpha were similar between CL responses stimulated with OZ and PMA. The increase of LgCL might be due to the inhibitory effect of IFN-alpha on myeloperoxidase degranulation, in view of the decreased LmCL responses.

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Measurement of chemiluminescence from neutrophils in a 96-well microplate using Lumi Box U-800 II

Cited by 30 publications

References 10 publications

Depressed humoral immunity after weight reduction in competitive judoists

Depressed humoral immunity after weight reduction in competitive judoists

Assessment of training effects on activity levels of alveolar macrophage in matured rats using chemiluminescent technique

Effect of interferon‐α on production of reactive oxygen species by human neutrophils

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