Chemiluminescent detection of induced reactive oxygen metabolite production of human polymorphonuclear leucocytes by anthophyllite asbestos

Our plain-English summary of Chemiluminescent detection of induced reactive oxygen metabolite production of human polymorphonuclear leucocytes by anthophyllite asbestos, by T. Iwata, N. Kohyama, E. Yano (Environmental Research, 2002).

The study

• Authors: T. Iwata, N. Kohyama, E. Yano
• Journal: Environmental Research
• Published: 2002;88(1):36-40
• Cited by: 4 articles, as counted by NIH iCite
• Read it: PubMed · Publisher

Abstract

Incidences of lung cancer and pleural plaque have been reported in relation to exposure to anthophyllite asbestos. To investigate the pathogenic mechanisms of anthophyllite, chemiluminescence (CL) detection of reactive oxygen metabolite (ROM) generation of human polymorphonuclear leucocytes (PMN) stimulated by anthophyllite asbestos was determined and compared with that of other asbestos and mineral fiber samples. When anthophyllite fiber sample was mixed with the luminol-primed PMN, high levels of CL which exhibited a specific time course characterized by two separate peaks were induced. The CL induced by anthophyllite sample was greater than that induced by chrysotile, crocidolite, and amosite asbestos. We further investigated the two peaks of CL using specific inhibitors of signal transduction mechanisms. The two peaks of CL by anthophyllite sample were different in sensitivity to cytochalasin B and genistein; the former relates to the cytoskeleton-dependent mechanism and the latter has been shown to inhibit tyrosine kinase, which resides in the pathway to cause PMN activation. The strong ROM reaction of PMN by anthophyllite suggests that the surface characteristics of the fiber may participate in the pathogenic mechanisms of anthophyllite asbestos.

In plain English

This was a laboratory experiment on human white blood cells (polymorphonuclear leucocytes, or PMN) taken from blood. The authors measured light given off by the cells (chemiluminescence) as a sign of reactive oxygen production after adding anthophyllite asbestos, chrysotile B, amosite, crocidolite or crystalline silica, each at 0.4 mg/ml. Results were checked in at least three separate experiments and shown as typical traces. Anthophyllite produced a sharp peak at about 1 minute and a broader peak at 5 minutes. Chrysotile B, amosite and crocidolite produced a broad peak at about 4 minutes, at a much lower level than anthophyllite or silica, although the chrysotile B response kept increasing even after 20 minutes. In tests with chemical inhibitors, cytochalasin B at 0.3 micrograms per ml suppressed the first anthophyllite peak while the second stayed almost unchanged. Genistein at 3 micromolar markedly suppressed the second peak while the first was unaffected. The authors concluded that the two peaks are generated by distinct biochemical mechanisms. They suggested that the fiber's surface characteristics may take part in the disease-causing mechanisms of anthophyllite asbestos.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory (in vitro) experiment measuring luminol-dependent chemiluminescence from human white blood cells exposed to mineral fibers, with and without signal-transduction inhibitors
• Who was studied: Human polymorphonuclear leucocytes (PMN) isolated from heparinized human blood. The number of blood donors is not stated. Each finding was reproduced in a minimum of three separate experiments, and a typical set of traces is shown in each figure.
• Exposure: UICC reference samples of chrysotile B, crocidolite, amosite and anthophyllite, plus a standard crystalline silica sample (min-u-sil), each added at a final concentration of 0.4 mg/ml. Inhibitors tested on the anthophyllite reaction were cytochalasin B, genistein, staurosporin and wortmannin.
• Main results:
  • Anthophyllite (0.4 mg/ml): chemiluminescence rose after a latent period of several seconds, with a sharp peak at about 1 minute followed by a broader peak at 5 minutes
  • Crystalline silica: a similar initial high peak with a shoulder at 2 to 3 minutes, but no distinct second peak
  • Chrysotile B, amosite and crocidolite: a broad peak at about 4 minutes at a much lower level than anthophyllite or crystalline silica
  • Chrysotile B-induced chemiluminescence continued to increase even after 20 minutes
  • Cytochalasin B at 0.3 micrograms per ml suppressed the first anthophyllite-induced peak, while the second broad peak remained almost unchanged. At the highest dose (5 micrograms per ml), 96% of cells were viable, compared with 98% in controls.
  • Genistein markedly suppressed the second anthophyllite-induced peak at the lowest dose of 3 micromolar, while the first peak was unaffected. At 50 micromolar the second peak was almost completely suppressed, and 96% of cells were viable.
  • Staurosporin at 0.25 micromolar slightly increased the first peak and decreased the second. At 0.5 micromolar both peaks were similarly decreased, and at 2 micromolar both were completely suppressed.
  • Wortmannin suppressed the first peak at 25 nM, with almost complete suppression at 200 nM. The authors observed no clear difference between the two peaks in sensitivity to staurosporin or wortmannin.
  • Anthophyllite from Afghanistan and from Yamaga (Japan) also induced similar levels and time courses of chemiluminescence. The authors did not show these data.
• The authors' conclusion: The authors concluded that anthophyllite induces a strong chemiluminescence reaction in PMN with two distinct peaks that differ in sensitivity to inhibitors, indicating that the two peaks are generated by distinct biochemical mechanisms. In their abstract they suggested that the fiber's surface characteristics may participate in the disease-causing mechanisms of anthophyllite asbestos.

Topics

Medical terms in this study, explained in our Lexicon: Anthophyllite, Lung Cancer, Pleural Plaques, Mineral Fiber, Chrysotile, Crocidolite, Amosite.

Related studies

• Cellular mechanisms of reactive oxygen metabolite generation from human polymorphonuclear leukocytes induced by crocidolite asbestos — T. Ishizaki, E. Yano, et al. (1997)
• Production of reactive oxygen metabolites induced by asbestos fibres in human polymorphonuclear leucocytes — M. Hedenborg, M. Klockars (1987)
• IgG and IgA enhance the chrysotile-induced production of reactive oxygen metabolites by human polymorphonuclear leucocytes — M. Klockars, M. Hedenborg, et al. (1989)
• Crocidolite-induced reactive oxygen metabolites generation from human polymorphonuclear leukocytes — T. Ishizaki, E. Yano, et al. (1994)
• Production of reactive oxygen species by man-made vitreous fibres in human polymorphonuclear leukocytes — M. Ruotsalainen, M.R. Hirvonen, et al. (1999)
• Changes in free intracellular calcium and production of reactive oxygen metabolites in human leukocytes by soluble and particulate stimuli — M. Tuomala, M.R. Hirvonen, et al. (1993)

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