Ndubuisi M I, Patel K, Rayanade R J, Mittelman A, May L T, Sehgal P B
Department of Cell Biology and Anatomy, New York Medical College, Valhalla 10514, USA.
J Immunol. 1998 Jan 1;160(1):494-501.
Transport of IL-6 in blood is fundamental to the biology of this cytokine. In the present study, IL-6 transport, immunological reactivity, and biological availability were investigated in blood from melanoma patients subjected to different active specific immunization regimens (an anti-idiotypic mAb immunization protocol (mAb-keyhole limpet hemocyanin (KLH)-Calmette-Guerin bacillus (BCG), an autologous anti-cancer vaccine protocol (AAAP), or both). Sera were subjected to Sephadex G-200 gel filtration chromatography, and the structure and biological activity of IL-6 complexes in the eluate fractions were probed using five IL-6 ELISAs and two bioassays. Sera from patients administered mAb-KLH+BCG followed by AAAP contained three distinct classes of IL-6 eluting at 30, 200, and 450 kDa, each with its characteristic ELISA reactivity and bioactivity: the 30- and 450-kDa complexes were bioactive in the B9 and Hep3B assays, but the 200-kDa complex was not. The 30- and 450-kDa IL-6 complexes were preferentially reactive in the 7IL6/5IL6 ELISA, the 200-kDa IL-6 complexes were preferentially reactive in the 4IL6/5IL6 ELISA, while the three commercial ELISAs (R&D, Endogen, and Genzyme) detected essentially only the 30-kDa IL-6. In contrast, 1) sera from AAAP patients contained biologically active 30- and 450-kDa IL-6 complexes, while 2) sera from mAb-KLH+BCG patients contained 200-kDa IL-6 complexes inactive in ex vivo bioassays. Both the 450- and 200-kDa complexes included soluble IL-6R, with the 200-kDa complexes additionally containing ligand-occupied anti-IL-6 and anti-soluble IL-6R IgG. The data indicate the existence of specific mechanisms that regulate the transport and function of IL-6 in vivo.
白细胞介素-6（IL-6）在血液中的运输对这种细胞因子的生物学特性至关重要。在本研究中，对接受不同主动特异性免疫方案（抗独特型单克隆抗体免疫方案（单克隆抗体-钥孔戚血蓝蛋白（KLH）-卡介苗（BCG））、自体抗癌疫苗方案（AAAP）或两者）的黑色素瘤患者血液中的IL-6运输、免疫反应性和生物学可用性进行了研究。血清进行了Sephadex G-200凝胶过滤色谱分析，并用五种IL-6酶联免疫吸附测定法（ELISA）和两种生物测定法检测洗脱级分中IL-6复合物的结构和生物学活性。接受单克隆抗体-KLH+BCG然后接受AAAP治疗的患者血清中含有三类不同的IL-6，分别在30、200和450 kDa处洗脱，每类都有其特征性的ELISA反应性和生物活性：30 kDa和450 kDa的复合物在B9和Hep3B测定中有生物活性，但200 kDa的复合物没有。30 kDa和450 kDa的IL-6复合物在7IL6/5IL6 ELISA中优先反应，200 kDa的IL-6复合物在4IL6/5IL6 ELISA中优先反应，而三种商业ELISA（R&D、Endogen和Genzyme）基本上只检测到30 kDa的IL-6。相比之下，1）AAAP患者的血清中含有具有生物活性的30 kDa和450 kDa的IL-6复合物，而2）单克隆抗体-KLH+BCG患者的血清中含有在体外生物测定中无活性的200 kDa的IL-6复合物。450 kDa和200 kDa的复合物都包含可溶性IL-6受体，200 kDa的复合物还额外含有与配体结合的抗IL-6和抗可溶性IL-6R IgG。数据表明存在调节体内IL-6运输和功能的特定机制。
