Torcia M, Lucibello M, Vannier E, Fabiani S, Miliani A, Guidi G, Spada O, Dower S K, Sims J E, Shaw A R, Dinarello C A, Garaci E, Cozzolino F
Department of Experimental Medicine, University of Roma Tor Vergata, Italy.
Exp Hematol. 1996 Jul;24(8):868-74.
We have studied the effects of several interleukin-1 (IL-1) inhibitors--IL-1 receptor antagonist (IL-1ra), soluble IL-1 receptor (sIL-1R) types I and II, and neutralizing monoclonal antibody (mAb) specific for IL-1 receptor type I--on the osteoclast-activating factor (OAF) activity of recombinant IL-1beta and of culture supernatants of unfractionated bone marrow mononuclear cells from multiple myeloma (MM) patients. The latter activity sharply correlated with the IL-1 content of culture supernatants (r = 0.949; p < 0.001). IL-1ra and sIL-1R types I and II had a clear-cut modulating effect on the OAF activity of IL-1beta at saturating doses (2-10 ng/mL); their effect was evident at 2 ng/mL and was dose-dependent over a large range of concentrations. Similarly, the three reagents neutralized the OAF activities of all MM cell supernatants in a dose-dependent fashion and completely abolished them when tested at the fixed concentration of 5 nM. The bone-resorbing activity of tumor necrosis factor-alpha (TNF-alpha) or lymphotoxin (LT), tested alone or added to MM cell supernatants, was affected not at all by IL-1ra and only minimally by sIL-1R types I and II, suggesting that little or no endogenous IL-1 was produced by the rat cells in the assay under TNF-alpha or LT stimulation. Consistent with these findings, PGE2 production elicited by IL-1beta or IL-1-rich supernatants in the rat long-bone assay was abolished by each reagent. Also, mAbs to the IL-1R p80 (type I) chains could modulate the effects of IL-1--recombinant or plasma cell-derived--in the OAF assay, but their activity was markedly less pronounced when compared with the IL-1 inhibitors, since they could never completely abolish bone resorption. Taken together, these findings demonstrate that inhibition of IL-1 interaction with cognate surface receptors on bone cells effectively counteracts its biologic activity. The findings also strongly indicate that OAF activity in conditioned medium of unfractionated myeloma bone marrow cells is predominantly, if not solely, related to IL-1beta.
我们研究了几种白细胞介素-1（IL-1）抑制剂——IL-1受体拮抗剂（IL-1ra）、I型和II型可溶性IL-1受体（sIL-1R）以及针对I型IL-1受体的中和单克隆抗体（mAb）——对重组IL-1β以及多发性骨髓瘤（MM）患者未分级骨髓单个核细胞培养上清液的破骨细胞激活因子（OAF）活性的影响。后者的活性与培养上清液中的IL-1含量显著相关（r = 0.949；p < 0.001）。在饱和剂量（2 - 10 ng/mL）下，IL-1ra以及I型和II型sIL-1R对IL-1β的OAF活性具有明显的调节作用；在2 ng/mL时其作用就很明显，并且在很大浓度范围内呈剂量依赖性。同样，这三种试剂以剂量依赖性方式中和了所有MM细胞上清液的OAF活性，当以5 nM的固定浓度进行测试时，能完全消除其活性。单独测试或添加到MM细胞上清液中的肿瘤坏死因子-α（TNF-α）或淋巴毒素（LT）的骨吸收活性，完全不受IL-1ra影响，仅受到I型和II型sIL-1R的轻微影响，这表明在TNF-α或LT刺激下，实验中的大鼠细胞几乎不产生或不产生内源性IL-1。与这些发现一致，在大鼠长骨实验中，每种试剂都消除了由IL-1β或富含IL-1的上清液引发的前列腺素E2的产生。此外，针对IL-1R p80（I型）链的单克隆抗体在OAF实验中可以调节IL-1（重组的或浆细胞衍生的）的作用，但其活性与IL-1抑制剂相比明显较弱，因为它们永远无法完全消除骨吸收。综上所述，这些发现表明抑制IL-1与骨细胞上同源表面受体的相互作用可有效抵消其生物学活性。这些发现还强烈表明，未分级骨髓瘤骨髓细胞条件培养基中的OAF活性如果不是完全由IL-1β引起，也是主要与之相关。
