Claassen E, Adler L T, Adler F L
J Histochem Cytochem. 1986 Aug;34(8):989-94. doi: 10.1177/34.8.2426338.
After incubation of tissue sections with anti-allotype-enzyme conjugates, the localization of immunoglobulin-allotype-bearing cells in the lymphoid tissues of conventional and chimeric rabbits could be established. The use of anti-allotype sera bearing distinct enzyme labels allowed simultaneous recognition of B cells producing immunoglobulin of one or the other parental types in heterozygous rabbits, or of B cells from the donor and recipient in chimeras. After immunization of chimeric rabbits with trinitrophenyl-keyhole limpet hemocyanin, anti-trinitrophenyl antibody-forming cells could be demonstrated through the use of a trinitrophenyl-alkaline phosphatase conjugate. Simultaneous incubation of sections with this reagent and with horseradish peroxidase coupled to (donor or recipient) anti-allotype sera made possible the determination of the origin (donor or recipient) of the antibody-forming cells. In agreement with the results of plaque assays and analyses of serum antibodies, all the anti-TNP producing cells were of donor origin when the chimeras had been created through injection of spleen or lymph node cells from trinitrophenyl primed donors. With this study we introduce a simple, direct method for the simultaneous identification of cells that produce antibody of a given allotype and a given specificity, applicable to appropriate studies in heterozygous or chimeric rabbits. The procedure has various advantages over previously reported methods.
将组织切片与抗同种异型酶结合物孵育后，可确定常规兔和嵌合兔淋巴组织中携带免疫球蛋白同种异型的细胞的定位。使用带有不同酶标记的抗同种异型血清能够同时识别杂合兔中产生一种或另一种亲代类型免疫球蛋白的B细胞，或嵌合体中来自供体和受体的B细胞。在用三硝基苯基-钥孔戚血蓝蛋白免疫嵌合兔后，通过使用三硝基苯基-碱性磷酸酶结合物可证明抗三硝基苯基抗体形成细胞。将切片同时与该试剂以及与（供体或受体）抗同种异型血清偶联的辣根过氧化物酶一起孵育，使得确定抗体形成细胞的来源（供体或受体）成为可能。与空斑试验结果和血清抗体分析一致，当通过注射来自三硝基苯基致敏供体的脾细胞或淋巴结细胞产生嵌合体时，所有抗TNP产生细胞均来自供体。通过这项研究，我们引入了一种简单、直接的方法，用于同时鉴定产生给定同种异型和给定特异性抗体的细胞，适用于对杂合兔或嵌合兔的相关研究。该方法相对于先前报道的方法具有多种优势。
