Rat interleukin-1 beta binding sites in rat hypothalamus and pituitary gland

In this study, radiolabeled recombinant rat interleukin-1 beta (r125I-IL-1 beta) was used to localize and characterize IL-1 beta binding in rat hypothalamus and pituitary gland by quantitative autoradiography. The ability of this ligand to bind to type I IL-1 receptor was first tested on murine lymphoma cells (EL-4). In the rat-tissue sections, high densities of specific r125I-IL-1 beta binding sites were localized in the anterior as well as the posterior pituitary and in the choroid plexus. A fine labeling was observed in meninges and third ventricle walls while no binding was detected in the hypothalamic nuclei. Saturation experiments, in the anterior and posterior pituitary, revealed one specific binding site with an affinity constant (Kd) of 0.5 nM. Competition experiments were achieved using either rat IL-1 beta (rIL-1 beta) or human IL-1s (hIL-1 alpha, hIL-1 beta and IL-1 receptor antagonist: hIL-1a). Affinity constants (Ki) were drastically different according to the ligand used, while Ki values were found similar in anterior and posterior pituitary. Competition with rIL-1 beta revealed one binding affinity (Ki of 0.1 nM range). In contrast, competition with hIL-1 beta revealed two binding affinities: a high (Ki: 0.1 pM range) and a low one (Ki: 1 nM range). Competition with hIL-1ra was obtained for high concentrations only (Ki: 10-100 nM range), whereas human IL-1 alpha (hIL-1 alpha) was unable to compete at 1-100 nM.(ABSTRACT TRUNCATED AT 250 WORDS)