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Research Article

Chemiluminescence in Human Whole Blood: Modulation by the Cocarcinogens Phorbol Diester and Polychlorobiphenyls

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Pages 417-435 | Published online: 28 Sep 2008
 

Abstract

A human whole blood chemiluminescence (CL) assay was established using zymosan as cell activator. Aroclor 1254 was found to inhibit this CL response in a direct linear relation to its concentration, (50% inhibitory dose, (ID50) equal to 5 × 10−4 M) in diluted blood samples of 10 normal human subjects. In comparison the ID50 of other inhibitors was 1.3 × 10−3 M for ethylenediamine tetraacetic acid, 3.3 × 10−3 M for ascorbic acid, 4 × 10−3 M for reduced glutathione, 1.2 × 10−3M for ethanol, 2.5 × 10−1 for methanol and 3.7 × 10−1 M for dimethyl sulfoxide. Using 12-o-tetradecanoyl-phorbol-13-acetate (TPA) as cell activator the CL response was likewise inhibited by Aroclor 1254 with an ID50 of 4.5 × 10−4 M. However, it was found that Aroclor 1254 alone has a stimulatory CL effect on otherwise unactivated cells. To compare the mechanisms involved in the CL elicited by the three stimulants zymosan, TPA and Aroclor 1254, the CL signal was measured in the presence of cytochalasin B. Cytochalasin B inhibited zymosan-induced CL, had a smaller inhibitory effect on TPA-induced CL but it could augment the CL response initiated by Aroclor 1254. This pattern of responses implicates Aroclor 1254 in the activation of eicosanoid metabolism as it matches the differential responses reported for arachidonic acid.

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