Interleukin 1 (IL-1) is a name that designates two proteins, IL-1?1and IL-1?, which are the products of distinct genes, but which recognize the same cell surface receptors. With the exception of skin keratinocytes, some epithelial cells, and certain cells of the central nervous system, IL-1 is not produced by the cells of healthy individuals. However, in response to stimuli such as those produced by inflammatory agents, infections, or microbial endotoxins, a dramatic increase in the production of IL-1 by macrophages and various other cell types is seen(1-3).
IL-1? 1and IL-1? are structurally related polypeptides that show approximately 25% homology at the amino acid level (2). Both are synthesized as 31 kDa precursors that are subsequently cleaved into proteins with molecular weights of approximately 17.5 kDa (4, 5).
Intracellular IL-1?consists exclusively of the 31 kDa precursor form (6). Extracellular IL-1? consists of a mixture of both unprocessed and mature IL-1?. These results indicate that processing takes place subsequently to secretion and is not tightly coupled to secretion (7). The specific protease apparently responsible for the processing of IL-1?, designated interleukin-1?-converting enzyme (ICE), has been described (7).
IL-1 possesses a wide variety of biological activities. IL-1 also plays an important role in immune functions, having effects on macrophages/ onocytes, T-lymphocytes, B lymphocytes, NK cells, and LAK cells. It acts on macrophages/ monocytes, inducing its own synthesis as well as the production of TNF and IL-6 (8,9). It activates T cells, resulting in IL-2 production and expression of IL-2 receptors (10). IL-1 also induces the production of GM-CSF and IL-4 from activated T cells (11). It induces B cell proliferation and maturation and increased immunoglobulin synthesis (12, 13).
For the quantitative determination of mouse interleukin 1 beta (IL-1?) concentrations in cell culture supernates, serum, and plasma.
Principle of the assay: This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-1? has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-1? present is bound by the immobilized antibody. Following incubation unbound samples are removed during a wash step, and then a detection antibody specific for IL-1? is added to the wells and binds to the combination of capture antibody-IL-1? in sample. Following a wash to remove any unbound combination, and enzyme conjugate is added to the wells. Following incubation and wash steps a substrate is added. A colored product is formed in proportion to the amount of IL-1? present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven IL-1? standard dilutions and IL-1? sample concentration determined.
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