Ochi H, Ohtsuka H, Yokota S, Uezumi I, Terashima M, Irie K, Noguchi H
Laboratory of Biotechnology, Takarazuka Research Center, Sumitomo Chemical Co., Ltd., Hyogo, Japan.
Infect Immun. 1991 Feb;59(2):550-4. doi: 10.1128/iai.59.2.550-554.1991.
Three stable hybridoma cell lines, IN-2A8, IN-5D6, and ZI-3A8, that secrete human monoclonal antibodies (MAbs) specific for b-type flagella of Pseudomonas aeruginosa were established by fusing peripheral blood lymphocytes from healthy volunteers with murine myeloma P3X63-Ag8.653 cells. The immunoglobulin M MAbs reacted specifically with flagellin (Mr, 52,000) by Western blotting (immunoblotting) analysis and bound specifically to clinical isolates belonging to Homma serotypes A, B, H, I, and M at frequencies of 58, 50, 46, 30, and 35%, respectively, but did not bind to any serotype E or G isolates. Overall, the MAbs bound to 31% of the clinical isolates. MAb IN-2A8 strongly protected burned mice challenged with P. aeruginosa bearing b-type flagella from death following parenteral administration of 0.1 microgram per mouse. This MAb also inhibited P. aeruginosa colony spreading in soft agar at a concentration of more than 1 microgram/ml but only slightly enhanced opsonophagocytosis by human polymorphonuclear leukocytes. A line of evidence suggests that the potent in vivo activity of MAb IN-2A8 in the burned-mouse model is likely to be caused by its inhibition of bacterial motility after binding to flagella.
通过将健康志愿者的外周血淋巴细胞与鼠骨髓瘤P3X63 - Ag8.653细胞融合，建立了三种稳定的杂交瘤细胞系IN - 2A8、IN - 5D6和ZI - 3A8，它们分泌对铜绿假单胞菌b型鞭毛具有特异性的人单克隆抗体（MAb）。通过蛋白质印迹（免疫印迹）分析，免疫球蛋白M单克隆抗体与鞭毛蛋白（分子量52,000）发生特异性反应，并分别以58%、50%、46%、30%和35%的频率特异性结合属于本间血清型A、B、H、I和M的临床分离株，但不与任何血清型E或G的分离株结合。总体而言，这些单克隆抗体与31%的临床分离株结合。单克隆抗体IN - 2A8在以每只小鼠0.1微克的剂量经肠胃外给药后，能强烈保护受到带有b型鞭毛的铜绿假单胞菌攻击的烧伤小鼠免于死亡。该单克隆抗体在浓度高于1微克/毫升时还能抑制铜绿假单胞菌在软琼脂中的菌落扩散，但仅略微增强人多形核白细胞的调理吞噬作用。一系列证据表明，单克隆抗体IN - 2A8在烧伤小鼠模型中的强大体内活性可能是由于其与鞭毛结合后抑制细菌运动所致。
