Locniskar M, Zumla A, Mudd D W, Isenberg D A, Williams W, McAdam K P
Department of Clinical Tropical Medicine, London School of Hygiene and Tropical Medicine, U.K.
Immunology. 1988 Jun;64(2):245-51.
Human monoclonal antibodies (mAb) were produced by hybridomas derived from fusion of the GM4672 lymphoblastoid cell line and peripheral blood mononuclear cells from leprosy patients. Hybridoma supernatants were screened for immunoglobulin (Ig) secretion, binding to Mycobacterium leprae, phenolic glycolipid-I (Phen GL-I), the unique M. leprae glycolipid and single-stranded(ss)DNA by ELISA. On the basis of direct-binding ELISAs, two IgMk mAb (PR4 and TH3) were selected for characterization. PR4 and TH3 bound to M. leprae, Phen GL-I and ssDNA; PR4 also bound to M. avium and M. kansasii and TH3 to M. kansasii. Inhibition assays demonstrated that these antibodies did not bind to the terminal disaccharide of Phen GL-I. In addition, both PR4 and TH3 bound to several autoantigens: ssDNA, double-stranded(ds)DNA and poly(ADP-ribose) but not RNA. PR4 and TH3 were used for preparation of rabbit anti-idiotype antisera. Inhibition studies demonstrated that the affinity purified rabbit anti-idiotype antisera were specific for their respective idiotype and that both Phen GL-I and ssDNA inhibited binding of idiotype to its anti-idiotype. PR4, but not TH3, was found to be similar but not identical to the 16/6 idiotype originally identified on a human monoclonal anti-DNA antibody derived from a patient with systemic lupus erythematosus (SLE).
人单克隆抗体（mAb）由GM4672淋巴母细胞系与麻风病人外周血单个核细胞融合产生的杂交瘤分泌。通过酶联免疫吸附测定（ELISA）筛选杂交瘤培养上清液，检测其免疫球蛋白（Ig）分泌情况以及与麻风分枝杆菌、酚糖脂-I（Phen GL-I）、麻风分枝杆菌独特的糖脂和单链（ss）DNA的结合能力。基于直接结合ELISA，选择了两种IgMκ mAb（PR4和TH3）进行特性鉴定。PR4和TH3与麻风分枝杆菌、Phen GL-I和ssDNA结合；PR4还与鸟分枝杆菌和堪萨斯分枝杆菌结合，TH3与堪萨斯分枝杆菌结合。抑制试验表明，这些抗体不与Phen GL-I的末端二糖结合。此外，PR4和TH3均与几种自身抗原结合：ssDNA、双链（ds）DNA和聚（ADP-核糖），但不与RNA结合。PR4和TH3用于制备兔抗独特型抗血清。抑制研究表明，亲和纯化的兔抗独特型抗血清对各自的独特型具有特异性，并且Phen GL-I和ssDNA均能抑制独特型与其抗独特型的结合。发现PR4与最初在一名系统性红斑狼疮（SLE）患者来源的人单克隆抗DNA抗体上鉴定出的16/6独特型相似但不完全相同，而TH3则不同。
