Interleukin 2 (IL-2) is a pleiotropic cytokine produced primarily by mitogenor antigen-activated T lymphocytes (1, 2). Human IL-2 (also known as T-cell growth factor) is produced by T-cells in response to antigenic or mitogenic stimulation. IL-2 is a potent lymphoid cell growth factor which exerts its biological activity primarily on T cells promoting proliferation and maturation.
Additionally, IL-2 has been found to stimulate growth and differentiation of B cells, NK cells, LAK cells, monocytes, and oligodendocytes. IL-2 is involved in treatment of cancers such as melanoma and renal cell cancer. It plays a key role in promoting the clonal expansion of antigen-specific T cells. In addition, IL-2 has also been shown to mediate multiple immune responses on a variety of cell types.
The sequence of mouse IL-2 cDNA predicts a 169 amino acid (aa) residue precursor glycoprotein containing a 20 aa residue signal peptide that is cleaved to form the mature protein (3, 4). At the amino acid sequence level, mature mouse IL-2 is approximately 60% identical to human IL-2 (1, 2, 5). Whereas human IL-2 is active on mouse cells, mouse IL-2 is species-specific and is inactive on human cells (6). The gene for IL-2 has been mapped to mouse chromosome 3 (7).
The biological effects of IL-2 are mediated by specific cell surface receptor complexes. The functional high-affinity receptor for IL-2 is composed of three distinct polypeptide chains (8, 9).
IL-2 stimulates the proliferation of thymocytes; stimulates the proliferation and differentiation of activated B cells; promotes the growth, differentiation and cytocidal activity of monocytes; induces the growth of natural killer cells and stimulates cytokine production by these cells as well as the cytolytic activity of these cells; enhances the production of lymphocyte-activated killer (LAK) cells; and induces the proliferation and differentiation of oligodendrocytes (1).
For the quantitative determination of mouse interleukin 2 (IL-2) 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-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-2 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-2 is added to the wells and binds to the combination of capture antibody-IL-2 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 coloured product is formed in proportion to the amount of IL-2 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-2 standard dilutions and IL-2 sample concentration determined.
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