Askonas B A, Williamson A R, Wright B E
Proc Natl Acad Sci U S A. 1970 Nov;67(3):1398-403. doi: 10.1073/pnas.67.3.1398.
A single antibody-forming cell clone has been selected from primed mice by sequential transfer of limited numbers of spleen cells into irradiated syngeneic mice. The original spleen cell donors had been immunized with dinitrophenylated bovine gamma globulin. Specific antibody molecules in sera of recipient mice were separated by isoelectric focusing on polyacrylamide gels and visualized by (131)I-hapten binding and autoradiography. This method provided a marker for antibody-forming cells derived from a single cell clone. This report describes the history of one clone of cells (E9) producing IgG antibody to dinitrophenyl. Clone E9 is long-lived and has been maintained for five transplant generations (over 6 months) by serial transfer of spleen cells into irradiated syngeneic mice. Clone E9 has the following properties: (1) Antibody production strictly depends on antigen, presented either in vivo or in vitro; (2) Induction of E9 anti-dinitrophenyl shows specificity for the carrier protein; (3) Antibody is produced in amounts (2-3 mg/ml serum) comparable with myeloma-protein production by murine plasmacytomas; (4) In the absence of antigen, memory cells have a lifetime exceeding 28 days.
通过将有限数量的脾细胞依次转移到经照射的同基因小鼠体内，从已致敏的小鼠中筛选出了一个单一的抗体形成细胞克隆。最初的脾细胞供体已用二硝基苯基化牛γ球蛋白进行了免疫。受体小鼠血清中的特异性抗体分子通过在聚丙烯酰胺凝胶上进行等电聚焦分离，并通过（131）I-半抗原结合和放射自显影进行可视化。该方法为源自单个细胞克隆的抗体形成细胞提供了一种标记。本报告描述了一个产生抗二硝基苯基IgG抗体的细胞克隆（E9）的历程。克隆E9寿命长，通过将脾细胞连续转移到经照射的同基因小鼠体内，已维持了五代移植（超过6个月）。克隆E9具有以下特性：（1）抗体产生严格依赖于抗原，无论是在体内还是体外呈现；（2）E9抗二硝基苯基的诱导对载体蛋白具有特异性；（3）产生的抗体量（2-3mg/ml血清）与小鼠浆细胞瘤产生的骨髓瘤蛋白量相当；（4）在没有抗原的情况下，记忆细胞的寿命超过28天。
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