Ortho-substituted polychlorinated biphenyls activate respiratory burst measured as luminol-amplified...

The effect of polychlorinated biphenyls (PCBs) on the activation of respiratory burst measured as luminol-amplified chemoluminescence in human granulocytes is elucidated here. Chemoluminescence was stimulated in a concentration-dependent manner (ED50 approximately 10 microM) by ortho-substituted PCB congeners, while meta- and para-substituted congeners had no significant effect. Two ortho-substituted PCB congeners were chosen for the mechanistic studies, namely 2,2',4,4'-TeCB and 2,2'-DCB, since they have been used in previous studies by others. In the absence of extracellular calcium, the respiratory burst in response to 2,2'-DCB and 2,2',4,4'-TeCB was reduced by 63% and 82%, respectively. Bisindolylmaleimide, which inhibits protein kinase C, reduced activated chemoluminescence by 2,2'-DCB, 2,2',4,4'-TeCB, N-formyl-methionyl-leucyl-phenylalanine, and phorbol 12-myristate 13-acetate. Neomycin, which inhibits phospholipase C, had a slight, but significant, effect on the 2,2',4,4'-TeCB-activated chemoluminescence but had a more pronounced effect on the 2,2'-DCB-activated chemoluminescence. 2,2'-DCB and 2,2',4,4'-TeCB significantly increased phospholipase D (PLD) activity measured as the amount of 14C-phosphatidylbutanol formed. Ethanol (1%), a phospholipase D modulator, reduced the response to 2,2'-DCB and 2,2',4,4'-TeCB by 72% and 75%, respectively. Furthermore, wortmannin (25 nM), a phosphatidylinositol 3-kinase, and genistein, a more unspecific tyrosine kinase inhibitor, reduced chemoluminescence in response to PCB. In conclusion, our results indicate that PCB-activated chemoluminescence is dependent on the Ca(2+)-dependent phospholipase D or phospholipase C, phosphatidylinositol 3-kinase, and protein kinase C activation prior to activation of the NADPH oxidase. Defects in neutrophhil functions upon exposure to PCB may render a greater susceptibility in the host to invading microorganisms or evoke inappropriate inflammatory responses leading to tissue injury.