Usuba O, Fujii Y, Miyoshi I, Naiki M, Sendo F
Department of Parasitology, Yamagata University School of Medicine.
Jpn J Cancer Res. 1988 Dec;79(12):1340-8. doi: 10.1111/j.1349-7006.1988.tb01565.x.
We have established a human-human hybridoma producing a monoclonal antibody against a tumor-associated carbohydrate-specific antigen, Hanganutziu-Deicher (HD) antigen. Human spleen lymphocytes from a patient with esophageal varices complicated with liver cirrhosis were cultured in serum-free medium and co-stimulated with both anti-human mu-chain antibodies and supernatants of concanavalin A-stimulated human spleen cell culture (ConA sup). The activated lymphocytes were subsequently primed in vitro with particulate HD3 antigen and fused with a parent hybrid myeloma cell line, KR-12. A hybridoma, 1F43E31G7 produced anti-HD human monoclonal antibody (IgM lambda). This monoclonal antibody reacted strongly with N-glycolylneuraminyl alpha 2-3 lactosylceramide (HD3) and slightly with N-glycolylneuraminyl alpha 2-3 lactoneotetraosylceramide (HD5), but did not react with N-glycolylneuraminyl alpha 2-3 lactoneohexaosylceramide (HD7), N-acetylneuraminyl alpha 2-3 lactosylceramide (GM3) and other derivatives of HD3 prepared by chemical modification of the sialic acid residue of HD3, which indicates that the monoclonal antibody is directed precisely toward the terminal sialic acid and whole structure of HD3.
我们建立了一种人-人杂交瘤,它能产生针对肿瘤相关碳水化合物特异性抗原汉加纽齐乌-戴歇尔(HD)抗原的单克隆抗体。从一名患有肝硬化并发食管静脉曲张的患者身上获取人脾淋巴细胞,在无血清培养基中培养,并与抗人μ链抗体和伴刀豆球蛋白A刺激的人脾细胞培养上清液(ConA sup)共同刺激。随后,活化的淋巴细胞在体外与颗粒状HD3抗原一起致敏,并与亲本杂交骨髓瘤细胞系KR-12融合。一种杂交瘤1F43E31G7产生了抗HD人单克隆抗体(IgM λ)。这种单克隆抗体与N-羟乙酰神经氨酰α2-3乳糖基神经酰胺(HD3)强烈反应,与N-羟乙酰神经氨酰α2-3乳糖新四糖基神经酰胺(HD5)轻微反应,但与N-羟乙酰神经氨酰α2-3乳糖新六糖基神经酰胺(HD7)、N-乙酰神经氨酰α2-3乳糖基神经酰胺(GM3)以及通过对HD3的唾液酸残基进行化学修饰制备的HD3的其他衍生物不反应,这表明该单克隆抗体精确地针对HD3的末端唾液酸和整个结构。