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Endotoxin-Stimulated Monocytes Release Multiple Forms of IL-1ß, Including a ProIL-1ß Form Whose Detection Is Affected by Export¹

Department of Pulmonary and Critical Care Medicine, Ohio State University, Columbus, OH 43210

The processing and release of 31-kDa proIL-1ß to the mature 17-kDa form of IL-1ß are still poorly understood. To help elucidate the mechanisms involved in IL-1ß processing and release, we measured IL-1ß forms released from endotoxin-stimulated monocytes by immunoprecipitation of [³⁵S]methionine-labeled protein, by Western blots, and by our recently developed ELISA specific for proIL-1ß. Our studies demonstrate that in addition to the 17-kDa mature IL-1ß, IL-1ß is also released as 31-, 28-, and 3-kDa molecules. The 31-kDa-released form of proIL-1ß represented 20–40% of the total released IL-1ß, as measured by SDS-PAGE with densitometry. This released proIL-1ß was susceptible to ICE processing; however, this proIL-1ß was not detectable by either a mature or proIL-1ß-specific ELISA, suggesting that release induces a conformational change. The ELISA inability to detect proIL-1ß was not due to inadequate sensitivity or subsequent degradation in the ELISA. Furthermore, while immunoaffinity-purified cytosolic proIL-1ß could complex the type II IL-1R, released proIL-1ß did not. Finally, the absence of a band shift in nondenaturing gel electrophoresis excluded proIL-1ß binding to another protein. These findings imply that IL-1ß is exported from monocytes as 3-, 17-, 28-, and 31-kDa forms and that the released 31-kDa form differs from cytosolic proIL-1ß.

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