Rolink A G, Melchers F, Palacios R
Basel Institute for Immunology, Switzerland.
J Exp Med. 1989 May 1;169(5):1693-701. doi: 10.1084/jem.169.5.1693.
The rat mAbs R52.120 and R52.625 inhibit the action of IL-5 on both IL-5-sensitive cell lines and freshly isolated splenic B lymphocytes. Neither antibody inhibits the proliferative cell responses promoted by IL-2, IL-3, or IL-4. Purified R52.120+ lymphoid spleen cells contain 15-20-fold higher numbers of B lymphocytes responding to IL-5 in the form of maturation into antibody-producing cells. By immunofluorescence staining and flow fluorocytometry, the R52.120 and R52.625 antibodies bound to all 12 IL-5-sensitive cell lines tested. Both antibodies react with 2-4% cells in the spleen, 5% lymphoid cells, and 10-15% myeloid cells in the bone marrow, and 10-14% in the peritoneum of C57BL/6, DBA/2, and BALB/c adult mice. No positive cells for either antibody were detected in the thymus and lymph nodes of these mice. Both R52.120 and R52.625 antibodies specifically inhibit the binding of radiolabeled IL-5 to its receptor. Finally, R52.120 and R52.625 antibodies precipitate from 35S-methionine-labeled IL-5-R+ cell lysates three proteins with Mr 46,000, 130,000, and 140,000. Taken together from these results, we conclude that the R52.120 and R52.625 mAbs recognize epitopes on the IL-5-R complex very close or identical to the IL-5 binding sites.
大鼠单克隆抗体R52.120和R52.625可抑制白细胞介素-5(IL-5)对IL-5敏感细胞系以及新鲜分离的脾B淋巴细胞的作用。这两种抗体均不抑制由IL-2、IL-3或IL-4促进的增殖性细胞反应。纯化后的R52.120 + 淋巴样脾细胞中,以成熟为抗体产生细胞的形式对IL-5产生反应的B淋巴细胞数量要高15至20倍。通过免疫荧光染色和流式荧光细胞术检测发现,R52.120和R52.625抗体与所有12种被测试的IL-5敏感细胞系均有结合。两种抗体与C57BL/6、DBA/2和BALB/c成年小鼠脾脏中的2-4%细胞、5%淋巴样细胞、骨髓中10-15%的髓样细胞以及腹膜中10-14%的细胞发生反应。在这些小鼠的胸腺和淋巴结中未检测到两种抗体的阳性细胞。R52.120和R52.625抗体均能特异性抑制放射性标记的IL-5与其受体的结合。最后,R52.120和R52.625抗体从35S-甲硫氨酸标记的IL-5-R + 细胞裂解物中沉淀出三种蛋白质,其分子量分别为46,000、130,000和140,000。综合这些结果,我们得出结论,R52.120和R52.625单克隆抗体识别的IL-5-R复合物表位与IL-5结合位点非常接近或相同。