<原著>ヒト好中球および単球における活性酸素産生能の基礎的および臨床的研究(1) : 活性酸素産生能測定法の基礎的検討
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Formation of oxygen radicals in human neutrophils and monocytes was investigated by performing the measurement of chemiluminescence (CL), the assessment of superoxide anion generation through the reduction of cytochrome c, and the application of flow cytometry (FCM) to assess oxidative products generated in phorbol myristate acetate (PMA)-stimulated neutrophils and monocytes as a novel attempt. CL in neutrophils was detected following stimulation with PMA, zymosan, opsonized zymosan, A23187,concanavalin A, n-formyl-methionyl-leucyl-phenylal-anine and aggregated IgG. Of these, PMA stimulation induced CL of 4.15×10^6 cpm, showing that it was the most effective method for CL. PMA-induced CL showed a severe reduction in neutrophils obtained from a male case of chronic granulomatous disease (CGD), namely 0.2% of the response observed in the control neutrophils. CL in his mother was also reduced to 28.4% of the control data after the stimulation with PMA. These results suggested that CL in neutrophils was a useful clue to the diagnosis or screening of CGD and its' carrier. Superoxide anion generation in neutrophils was measured by superoxide dismutase (SOD)-inhibitable cytochrome c reduction to evaluate other oxygen radical-induced responses than CL. In 6 healthy subjects, the values were 2.03±0.23 nmol/min/10^6 cells (mean±SE). The FCM technique was applied to monitor the formation of oxygen radicals in neutrophils or monocytes evaluated as cumulative fluorescence intensity (CFI). CFI observed in a healthy subject showed 172.9 in neutrophils and 24.9 in monocytes. The results of FCM showed a good correlation (r=0.92) with those of PMA-stimulated CL in neutrophils. Therefore, the two assays were considered to determine the same oxygen species. It was also suggested that FCM would be very useful to measure the oxidative products generated specifically in neutrophils or in monocytes. Furthermore a significant correlation was found between PMA-induced CL and % Fc receptor (FcR ) positive cells in neutrophil suspensions. FcR-negative cells showed low levels in both CL responsiveness to PMA and generation of oxidative products measured with FCM compared with the positive ones. These data indicate that FCR on neutrophils has a close relationship with the oxygen radical formation in them.
- 近畿大学の論文
- 1987-03-25
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