Environmental Toxicology and Pharmacology 5 (1998) 105 – 112 Ortho substituted polychlorinated biphenyls elevate intracellular [Ca 2 + ] in human granulocytes Ø.A. Voie *, F. Fonnum Norwegian Defence Research Establishment, Diision for Enironmental Toxicology, P.O. Box 25, N-2007, Kjeller, Norway Received 23 April 1997; received in revised form 8 October 1997; accepted 13 October 1997 Abstract The perturbation of Ca 2 + -homeostasis in human granulocytes exposed to ortho and non ortho substituted polychlorinated biphenyls (PCB) was investigated. Ortho substituted PCB congeners increased intracellular free calcium, [Ca 2 + ]i, in a concentra- tion-dependent manner. The increase in [Ca 2 + ]i was inversely proportional to the total surface area of the ortho substituted congeners. The effect of ortho substituted PCB congeners was dependent upon external Ca 2 + and phospholipase C activation, except for a tetra-ortho substituted congener, 2,2,6,6-TeCB, that was not phospholipase C-dependent. We suppose that PCBs activate phospholipase C which leads to the production of ins(1,4,5)P3. This will release Ca 2 + from intracellular stores and subsequently activation of Ca 2 + release activated Ca 2 + channels (CRAC) in the plasma membrane. It is also possible that PCBs activate CRAC in a more direct manner. Our findings show that ortho substituted PCB congeners stimulate [Ca 2 + ]i elevation in human granulocytes, and this could in part account for the effects of PCB on the immune system. © 1998 Elsevier Science B.V. All rights reserved. Keywords: Polychlorinated biphenyls (PCB); Granulocytes; Ca 2 + ; Protein kinase C; Phospholipase C 1. Introduction Polychlorinated biphenyls (PCB) are stable lipophilic compounds which possess a strong biomagnification ability and have become widespread environmental pol- lutants. PCBs have been used as isolating and cooling liquids in electrical components. These compounds show a complex spectrum of biological and toxicologi- cal properties (Van den Berg et al., 1993). Effects such as cytochrome P450 1A1 induction, hepatomegaly, thymic atrophy and reduced body weight gain are all mediated by coplanar PCB congeners through a com- mon receptor, the Ah receptor (Nebert, 1989; Safe, 1990). Little attention has been paid to di-, tri- and tetra-ortho PCB congeners. It has been shown that these congeners, which are abundantly present in the environment, cause other biological and toxic effects such as P450 2B induction, hepatotoxicity, carcinogen- ity, neurotoxicity and immunotoxicity (Parkinson et al., 1983; Davis and Safe, 1990; Silberhorn et al., 1990; Seegal et al., 1991). Toxic effects of PCBs on phago- cytic cells of experimental animals have been reported (Loose et al., 1978). Thomas and Hinsdill (1978), have found that exposure of mice to PCBs enhanced suscep- tibility to endotoxin shock, and reduced the ability of the animals to clear the injected microorganisms. A di-ortho PCB congener (2,2,4,4-TeCB) was recently found to alter neutrophil function by both stimulating degranulation and by superoxide production in rats (Brown and Ganey, 1995). Mature granulocytes consist of neutrophils (5 ×10 9 cells/l blood), eosinophils (2 × 10 8 cells/l blood) and basophils (4 ×10 7 cells/l blood) and are developed from a common pluripotent stem cell (Cline, 1975). To achieve its mission of localizing and destroying micro-organisms, the neutrophil must be * Corresponding author. Tel.: +47 63807824; fax: +47 63807811; e-mail: oyvind-albert.voie@ffi.no 1382-6689/98/$19.00 © 1998 Elsevier Science B.V. All rights reserved. PII S1382-6689(97)10064-3