Human Interleukin-1β Research Articles

Sandwich enzyme immunoassay for human interleukin-1β (hIL-1β) in urine

A highly sensitive sandwich enzyme immunoassay for human interleukin-1β (hIL-1β) in urine is described. Rabbit anti-hIL-1/gb IgG-coated polystyrene balls were incubated with urine samples and subsequently with affinity-purified rabbit anti-hIL-1/gb Fab'-horseradish peroxidase conjugate. After washing, the peroxidase activity bound to the polystyrene ball was assayed by fluorimetry using 3-(4-hydroxyphenyl) propionic acid as a substrate. The detection limit of hIL-1β was 0.5 pg/tube (30 amol/tube) or 5 ng/l of urine when 0.1 ml of urine sample was used. The molecular weight of hIL-1β in normal urine was shown to be 17000 by gel filtration, and levels of urine hIL-1β in healthy subjects aged 19–87 yr were 0–146 ng/l or 0–88 ng/g of creatinine.

In vitro production of human interleukin 1 alpha and interleukin 1 beta by peripheral blood mononuclear cells examined by sensitive sandwich enzyme immunoassay.

The in vitro production of human interleukin 1 alpha (hIL 1 alpha) and interleukin 1 beta (hIL 1 beta) by peripheral blood mononuclear cells was examined by sensitive sandwich enzyme immunoassays which could discriminate hIL 1 alpha and hIL 1 beta without cross-reaction with human IL2. In culture supernatants of mononuclear cells, two components were detected by sandwich enzyme immunoassay for hIL 1 alpha or hIL 1 beta. The molecular weight of one component was shown to be equal to that of recombinant hIL 1 alpha or hIL 1 beta by gel filtration. The elution volume of the other component corresponded to a molecular weight of about 30,000. The sum of the two components for both hIL 1 alpha and hIL 1 beta in culture supernatants of peripheral blood mononuclear cells from healthy subjects increased 1.7 to 38-fold by Escherichia coli lipopolysaccharide. The sum of the two components for hIL 1 beta was 13 to 97-fold larger than that for hIL 1 alpha.

Synthesis and Cloning of a Gene Encoding Human Interleukin-1β (IL-1β)

Abstract The design, synthesis and cloning of a 466 base-pair DNA duplex coding for IL-lp is described.

Stimulation of arachidonic acid metabolism: Differences in potencies of recombinant human interleukin-1α and interleukin-1β on two cell types

Recombinant human interleukin-1 (rIL-1) α and β, which have 26% homology in their amino acid sequence, stimulated arachidonic acid metabolism by squirrel monkey smooth muscle cells and rat liver cells; their relative effectiveness, however, varied with the two cells. Recombinant IL-1α was 3 times more effective than rIL-1β at stimulating arachidonic acid metabolism by the primate smooth muscle cells. Recombinant IL_1α was 3 time less effective than rIL-1β when measured by their capacity to synergistically stimulate arachidonic acid metabolism of rat liver cells in the presence of palytoxin and anti-diuretic hormone (ADH). The rIL-1α and rIL-1β also stimulated the release of radiolabelled arachidonic acid from the smooth muscle cells prelabelled with [ 3H]arachidonic acid. The two recombinant IL-1s have different heat stabilities, again when measured by their capacity to stimulate arachidonic acid metabolism; IL-1α was more heat stable than IL-1β.

A 1H-NMR study of human interleukin-1 beta. Sequence-specific assignment of aromatic residues using site-directed mutant proteins.

Complete identification of spin systems in the aromatic region of recombinant human interleukin-1 beta has been achieved using two-dimensional homonuclear Hartmann-Hahn spectroscopy. In addition, sequence-specific assignments for the four tyrosine residues have been carried out with the help of a series of mutant proteins, obtained by site-directed mutagenesis of the cloned gene. It is shown that, for the mutant proteins investigated, either none or only local structural changes occur. The use of NMR spectroscopy to determine the structural identity of site-directed mutant proteins with respect to the wild-type protein is discussed.