STUDY ON THE DEVELOP OF IDIOTYPIC ANTIBODY TO ESTROGEN AND ON THE APPLICATION IT TO CLINICAL USE

1974年，尼尔斯·杰恩提出了一种理论，将免疫系统描绘成一个相互作用的独特型网络。这种免疫反应的网络理论强调，需要将单个免疫球蛋白分子视为表达两个关键集合，它们通过这两个集合与免疫系统的其他元素相互作用。每个分子都有一个结合点，通过这个结合点，它可以与传统的抗原决定簇相互作用，也可以与其他免疫球蛋白分子相互作用，这些免疫球蛋白分子带有与抗原交叉反应的独特型决定簇。杰恩将免疫球蛋白的抗原结合部位指定为副表位。传统抗原的抗原决定簇称为表位。每个免疫球蛋白分子表达一个或多个由其可变区的结构决定的抗原决定因素，因此对它来说是独一无二的，或者至少只被相对有限数量的免疫球蛋白分子共享。这些独特型决定因素被称为独特位。在这些…的基础上在更多的情况下，我们试图制造与雌激素受体结合的独特型抗体，我们推测其中一些应该具有雌激素的生物活性。独特型抗体是分析细胞内雌激素受体的良好工具，是临床上有用的药物，是免疫球蛋白而不是类固醇结构。首先，我们制备了抗雌激素的多克隆抗体和单抗。然后将雌激素抗体注射到小鼠体内，产生独特型抗体，即抗雌激素免疫球蛋白。在多次加强免疫后，收集小鼠脾细胞与小鼠骨髓瘤细胞融合，制成产生独特型抗体的杂交瘤细胞。用酶联免疫吸附试验(ELISA法)和~3H-雌激素与雌激素抗体的结合抑制法检测其独特型活性。我们尝试了几种细胞融合，得到了数千个杂交瘤和100多个产生具有雌激素样活性物质的单抗细胞。我们进一步分析了这些物质的生物学、生化和化学性质。体内和体外的生物活性显示出雌激素样的活性，但生化分析表明该物质不是免疫球蛋白，是低相对分子质量的化学物质。结果表明，该杂交瘤细胞在体外具有合成雌激素的能力，这一结果令人振奋。较少

In 1974, Niels Jerne proposed a theory which pictured the immune system as a network of interacting idiotypes. This network theory of immune response emphasized the need to consider individual immunoglobulin molecules as expressing two critical sets through which they interacted with other elements of the immune system. Each molecule bears a combining site through which it interacts with conventional antigenic determinants and with other immunoglobulin molecules which bore idiotypic determinants cross-reacting with antigen. Jerne designated the antigen-combining sites of immunoglobulins as paratopes. The antigenic determinants of conventional antigens are referred to as epitopes. Each immunoglobulin molecule expresses one, or many, antigenic determinants determined by the structure of its variable regions and thus unique to it, or at least shared by only a relatively limited number of immunoglobulin molecules. These idiotypic determinants were termed as idiotopes. on the bases on these … More situation, we tried to make idiotypic antibodies that bind to estrogen receptor.We speculated that some of them should have biological activity as estrogen has. The idiotypic antibodies would be a good tool to analyze estrogen receptors in cells and be a useful medicine, which are immunoglobulin but not steroid structure, for clinical fields. First of all, we made polyclonal and monoclonal antibodies to estrogen. Then the estrogen antibodies were injected to mice to rise idiotypic antibody, that is, anti-anti-estrogen immunoglobulin. After giving several boosters, mouse spleen cells were collected and fused with mouse myeloma cells to make hybridoma producing the idiotypic antibody. The idiotypic activity was checked by the enzyme-linked immunosorbant assay (ELISA) and the binding inhibition of ^3H-estrogen to estrogen antibody. We tried several cell fusion and got thousands of hybridomas and more than one hundred monoclonal cells producing substances having estrogen like activity. We further more analyzed the biological, biochemical and chemical characteristics of these substances. The biological activity using in vivo and in vitro showed estrogen-like activity, however, biochemical analysis showed that the substance was not immunoglobulins and was lower molecular weight chemicals. We finally concluded that the substance was estrogen itself by HPLC and mass spectrography.These finding is very exciting one that high percentages of hybridoma have an ability to synthesize estrogen in vitro. Less