General methods for the study of cells and serum during the immune response: the response to dinitrophenyl in mice.
General methods for the study of cells and serum during the immune response: the response to dinitrophenyl in mice.
复制标题
研究免疫反应期间细胞和血清的一般方法：小鼠对二硝基苯基的反应。
DOI：
--
复制
发表时间：
1969
影响因子：
4.6
通讯作者：
R. B. Taylor
中科院分区：
文献类型：
--
作者：
N. Klinman;R. B. Taylor
本文报道了两种新的抗体测定方法：一种是测定溶液中的抗体，另一种是测定单个细胞释放的抗体。它们依赖于使用免疫球蛋白结合抗原来吸收抗体，并使用放射性标记的抗免疫球蛋白抗体（抗Ig）来检测抗体。在第一种方法中，将免疫吸附剂加入到含有抗体的溶液中，洗涤除去除结合抗体以外的血清组分，用[131 I]抗Ig处理，并通过γ射线计数评估放射性摄取。通过与平衡透析结果的比较，将该方法用于抗DNP抗体标准化。它被证明是独立的亲和力，低至K 0 = 106 1/M。可检测到0·1 ng的抗体。为了研究淋巴细胞释放抗体，将细胞和免疫吸附剂分散在显微镜载玻片上的琼脂糖凝胶中，在[125 I]抗Ig存在下于37°C下孵育，洗涤并放射自显影。然后，细胞释放的抗体可以被可视化为银颗粒的圆形区域（“点”）。当使用绵羊红细胞作为抗原时，斑点的数量接近于通过添加补体获得的裂解斑的数量。所述方法显示出对于许多抗原通常是有用的。这两种方法都用于研究小鼠对二硝基苯的初次免疫应答过程。免疫后第2天开始检测到抗DNP抗体释放细胞，第9天在注射部位局部淋巴结中达到峰值，为2 × 105。
Two new methods for antibody assay are reported: one for antibody in solution, the other for individual cells releasing antibody. They depend on the use of an immunoabsorbent-bound antigen to absorb the antibody, and radioactively-labelled anti-immunoglobulin antibody (anti-Ig) for its detection. In the first method immunoabsorbent was added to the solution containing antibody, washed free of serum components other than the bound antibody, treated with [131I]anti-Ig, and the uptake of radioactivity assessed by γ-ray counting. This method was standardized for anti-DNP antibody by comparison with results of equilibrium dialysis. It was shown to be independent of affinity, down to K0 = 106 1/M. It would detect 0·1 ng antibody. For the study of lymphoid cells releasing antibody the cells and immunoabsorbent were dispersed in agarose gel on microscope slides, incubated at 37°C in the presence of [125I]anti-Ig, washed, and autoradiographed. Antibody released by a cell could then be visualized as a circular area (`spot') of silver grains. When sheep red blood cells were used as the antigen, the number of spots approximated to the number of lytic plaques obtained by the addition of complement. The methods were shown to be generally useful for a number of antigens. Both methods were used in a study of the course of the primary immune response to dinitrophenyl in mice. Cells releasing anti-DNP antibody were detected from 2 days after immunization and rose to a peak number at 9 days of 2 × 105 in the lymph nodes local to the site of injection.