Barclay S L, Smith A M
Proc Natl Acad Sci U S A. 1986 Jun;83(12):4336-40. doi: 10.1073/pnas.83.12.4336.
Two immunization procedures were compared for their ability to yield monoclonal antibodies that react with plasma membrane-bound differentiation antigens of Dictyostelium. In the first method, hybridomas prepared from BALB/c mice immunized with aggregating amoebae produced monoclonal antibodies that recognized antigens present on both growing and aggregating Dictyostelium amoebae. None of the monoclonal antibodies reacted with only the injected aggregation-stage cell type. In contrast, monoclonal antibodies that reacted with differentiation antigens were easily obtained by primary immunization of BALB/c mice with living aggregation-stage cells, followed by secondary immunization with a preparation of plasma membrane from aggregating cells or intact aggregating cells mixed with polyclonal BALB/c antiserum raised against undifferentiated cells. By this method, approximately 20% of all anti-Dictyostelium monoclonal antibodies obtained in a fusion are specific for differentiation antigens. The properties and developmental regulation of several of these antigens are described. The possible uses of this immunological method to detect unique determinants on other kinds of cells and the likely immune mechanisms that make it successful are discussed.
比较了两种免疫程序产生与盘基网柄菌质膜结合分化抗原发生反应的单克隆抗体的能力。在第一种方法中，用聚集变形虫免疫BALB/c小鼠制备的杂交瘤产生了识别生长和聚集的盘基网柄菌变形虫上存在的抗原的单克隆抗体。没有一种单克隆抗体仅与注射的聚集期细胞类型发生反应。相比之下，通过用活的聚集期细胞对BALB/c小鼠进行初次免疫，然后用来自聚集细胞的质膜制剂或与针对未分化细胞产生的多克隆BALB/c抗血清混合的完整聚集细胞进行二次免疫，很容易获得与分化抗原发生反应的单克隆抗体。通过这种方法，在融合中获得的所有抗盘基网柄菌单克隆抗体中约20%对分化抗原具有特异性。描述了其中几种抗原的特性和发育调控。讨论了这种免疫方法用于检测其他类型细胞上独特决定簇的可能用途以及使其成功的可能免疫机制。
