Time course of superoxide generation by leukocytes-The MCLA chemiluminescence system

L. Prónai, Hiroe Nakazawa, Kohji Ichimori, Yoshinori Saigusa, Tomoichi Ohkubo, Kazuko Hiramatsu, Shigeru Arimori, J. Fehér

Research output: Contribution to journalArticle

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Abstract

This study was performed to examine the pattern of Superoxide (O2-·) generation from leukocytes using the O2-· specific chemiluminescence (CL) method. Cypridina luciferin analog, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-alpha]pyrazin-3-one (MCLA) was used as a CL probe. The appropriate conditions of the MCLA method was first determined for the evaluation of the time course of O2-· generation by leukocytes. The time course of O2-· generation obtained by the MCLA-CL system was compared with that by the luminol-dependent CL, electron spin resonance (ESR)/spin trapping, and cytochrome c systems. Following stimulation by three different stimulants (PMA, OZ, FMLP), leukocytes continuously generated O2-· for up to 5 h in the MCLA-CL system, irrespective of the kind of stimulation. The curves obtained by generation ceased more rapidly in the luminol-CL, ESR/spin trapping, and cytochrome c systems. A 50% activity of the initial value was observed at 70 min in the MCLA-CL system, but 30, 10 and 35 min in the other systems, respectively. The CL or O2-· generation value decreased to less than 1% (possible termination) at 300, 90, 120 and 180 min, respectively. With the exception of ESR studies with OZ, the cell viability was not significantly affected in any of the trials. These results indicate that leukocytes can generate O2-· much longer than previously estimated and that the MCLA-CL-system is the most suitable system for the measurement of the O2-· generation by leukocytes.

Original languageEnglish
Pages (from-to)437-450
Number of pages14
JournalInflammation
Volume16
Issue number5
DOIs
Publication statusPublished - Oct 1992

Fingerprint

Luminescence
Superoxides
Leukocytes
Electron Spin Resonance Spectroscopy
Spin Trapping
Luminol
Cytochromes c
2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one
Cell Survival

ASJC Scopus subject areas

  • Medicine(all)
  • Immunology
  • Cell Biology

Cite this

Time course of superoxide generation by leukocytes-The MCLA chemiluminescence system. / Prónai, L.; Nakazawa, Hiroe; Ichimori, Kohji; Saigusa, Yoshinori; Ohkubo, Tomoichi; Hiramatsu, Kazuko; Arimori, Shigeru; Fehér, J.

In: Inflammation, Vol. 16, No. 5, 10.1992, p. 437-450.

Research output: Contribution to journalArticle

Prónai, L, Nakazawa, H, Ichimori, K, Saigusa, Y, Ohkubo, T, Hiramatsu, K, Arimori, S & Fehér, J 1992, 'Time course of superoxide generation by leukocytes-The MCLA chemiluminescence system', Inflammation, vol. 16, no. 5, pp. 437-450. https://doi.org/10.1007/BF00918970
Prónai L, Nakazawa H, Ichimori K, Saigusa Y, Ohkubo T, Hiramatsu K et al. Time course of superoxide generation by leukocytes-The MCLA chemiluminescence system. Inflammation. 1992 Oct;16(5):437-450. https://doi.org/10.1007/BF00918970
Prónai, L. ; Nakazawa, Hiroe ; Ichimori, Kohji ; Saigusa, Yoshinori ; Ohkubo, Tomoichi ; Hiramatsu, Kazuko ; Arimori, Shigeru ; Fehér, J. / Time course of superoxide generation by leukocytes-The MCLA chemiluminescence system. In: Inflammation. 1992 ; Vol. 16, No. 5. pp. 437-450.
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