Oxidative inactivation of leukotriene C4 by stimulated human polymorphonuclear leukocytes

Leukotriene C(4) (LTC(4)) was metabolized by human polymorphonuclear leukocytes (PMNs) stimulated with phorbol myristate acetate (PMA) into three sets of products. These products differed in mobility on reverse-phase high-performance liquid chromatography (RP HPLC) from LTC(4) and also from leukotriene D(4) (LTD(4)) and leukotriene E(4) (LTE(4)), the sequential products of peptide cleavage of LTC(4). Products I, II, and III were eluted as doublets with an average retention time for each doublet of 7.5 +/- 0.3, 10.5 +/- 0.6, and 16.3 +/- 1.1 min (mean +/- SD), respectively, as compared with 13.8 min for LTC(4). Doublet I material was biologically inactive and showed <5% of the immunoreactivity of LTC(4), doublet II material had 1% of the spasmogenic activity of LTC(4) on the guinea pig ileum and was equally immunoreactive, and doublet III material was neither biologically active nor immunoreactive. When [14,15-(3)H]LTC(4) and [(35)S]LTC(4) were metabolized, all three doublet products retained the (3)H label, whereas only the doublet I and doublet II products retained the (35)S label. The UV absorbance spectra of the three sets of metabolites were as follows: doublet I, maximum at 280 nm with shoulders at about 270 and 290 nm; doublet II, maximum at 284.5 nm with shoulders at about 275 and 295 nm; and doublet III, maximum at 269 nm with shoulders at about 259 and 279 nm. The metabolism of LTC(4) to the three classes of functionally inactive products by stimulated PMNs was completely blocked by catalase and azide, indicating a requirement for H(2)O(2) and myeloperoxidase. When hypochlorous acid (HOCl)-considered to be a natural product of the interaction of myeloperoxidase, H(2)O(2), and chloride ion-was formed chemically and allowed to react with LTC(4), the resulting products were indistinguishable by UV and HPLC analyses from the doublet II and doublet III metabolites of LTC(4). The doublet II products were identified as the two diastereoisomeric sulfoxides of LTC(4) by comparison with synthetic reference compounds. The doublet III products were shown to be identical with synthetic samples of (5S, 12S)- and (5S, 12R)-6-trans-LTB(4). The formation of two diastereoisomeric LTC(4) sulfoxides and 6-trans-LTB(4) can be explained in terms of an S-chlorosulfonium ion as the initial reactive intermediate, which subsequently undergoes conversion to product II by hydrolysis and product III by carbocation formation.