Buchner R R, Vogen S M, Fischer W, Thoman M L, Sanderson S D, Morgan E L
Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037, USA.
J Immunol. 1997 Feb 15;158(4):1670-80.
Site-directed polyclonal Abs specific for a synthetic peptide with sequence homology to the predicted N-terminal sequence of the human kappa opioid receptor [anti-kappa R-(33-52)] are capable of binding to normal human cells and cell lines expressing mRNA specific for the human kappa receptor. Flow cytometric analysis of 1) a neuronal cell line (NT2), 2) blood-derived CD14+ monocytes, 3) monocyte-like cell lines (U937 and THP 1), 4) blood-derived CD3+ T cells and a T cell line, and 5) human B cell lines bound anti-kappa R-(33-52) in a specific manner. Anti-kappa R-(33-52) was also found to specifically neutralize the immunosuppressive activities associated with the kappa R-selective agonist U50,488H. This antiserum was found to block U50,488H-mediated inhibition of 1) Staphylococcus aureus Cowen strain I-induced B and T lymphocyte proliferation, 2) PHA-induced T lymphocyte proliferation, and 3) S. aureus Cowen strain I-induced IgG production. However, this antiserum failed to neutralize mu R-selective agonist (Tyr-D-Ala-Gly-NMe-Phe-Gly-ol)-mediated suppression of IgG synthesis. Finally, the kappa R-selective antagonist nor-binaltorphimine hydrochloride inhibits the binding of anti-kappa R-(33-52) to the U937 cell line. These results suggest that anti-kappa R-(33-52) specifically interacts with the human kappa R molecule. Studies conducted with anti-kappa R-(33-52) indicated that this antiserum effectively blocked U50,488H-mediated immunosuppression, but by itself did not enhance or suppress lymphocyte activation. These data suggest that anti-kappa R-(33-52) 1) does not interact with the effector binding site of the receptor, but sterically interferes with U50,488H binding to the receptor; and/or 2) the antiserum interacts with a secondary binding site that is important for ligand binding, but may not be involved in signal transduction.
针对一种与人类κ阿片受体预测N端序列具有序列同源性的合成肽的位点特异性多克隆抗体[抗κR-(33-52)]，能够与表达人类κ受体特异性mRNA的正常人细胞和细胞系结合。对以下细胞进行流式细胞术分析：1) 一种神经细胞系(NT2)、2) 血液来源的CD14+单核细胞、3) 单核细胞样细胞系(U937和THP 1)、4) 血液来源的CD3+ T细胞和一种T细胞系，以及5) 人类B细胞系，结果显示抗κR-(33-52)以特异性方式与之结合。还发现抗κR-(33-52)能特异性中和与κR选择性激动剂U50,488H相关的免疫抑制活性。该抗血清能阻断U50,488H介导的以下抑制作用：1) 金黄色葡萄球菌考恩I株诱导的B和T淋巴细胞增殖、2) 植物血凝素诱导的T淋巴细胞增殖、3) 金黄色葡萄球菌考恩I株诱导的IgG产生。然而，该抗血清未能中和μR选择性激动剂(酪氨酰-D-丙氨酰-甘氨酰-N-甲基苯丙氨酰-甘氨醇)介导的IgG合成抑制作用。最后，κR选择性拮抗剂盐酸去甲二氢吗啡酮抑制抗κR-(33-52)与U937细胞系的结合。这些结果表明抗κR-(33-52)与人κR分子发生特异性相互作用。用抗κR-(33-52)进行的研究表明，该抗血清有效阻断了U50,488H介导的免疫抑制作用，但自身并未增强或抑制淋巴细胞活化。这些数据表明抗κR-(33-52)：1) 不与受体的效应器结合位点相互作用，而是在空间上干扰U50,488H与受体的结合；和/或2) 该抗血清与对配体结合很重要的二级结合位点相互作用，但可能不参与信号转导。
