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

1974年，Niels Jerne提出了一个理论，将免疫系统描绘为一个相互作用的独特型网络。免疫反应的网络理论强调需要考虑单个免疫球蛋白分子表达两个关键集合，通过它们与免疫系统的其他元素相互作用。每个分子带有一个结合位点，通过该位点它与常规抗原决定簇和具有与抗原交叉反应的独特型决定簇的其他免疫球蛋白分子相互作用。Jerne将免疫球蛋白的抗原结合位点称为互补位。常规抗原的抗原决定簇被称为表位。每个免疫球蛋白分子表达一个或多个抗原决定簇，这些抗原决定簇由其可变区的结构决定，因此是其独有的，或者至少仅由相对有限数量的免疫球蛋白分子共享。这些独特型决定子被称为独特位。在这些基础上 ...更多信息 在这种情况下，我们试图制造与雌激素受体结合的独特型抗体，我们推测其中一些抗体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，是一种免疫球蛋白而非甾体结构的药物，可用于临床。首先，我们制备了雌激素的多克隆抗体和单克隆抗体。然后将雌激素抗体注射到小鼠体内，产生独特型抗体，即抗抗雌激素免疫球蛋白。在给予几次加强剂后，收集小鼠脾细胞并与小鼠骨髓瘤细胞融合以制备产生独特型抗体的杂交瘤。用酶联免疫吸附试验（ELISA）和^3H-雌激素对雌激素抗体结合抑制试验检测其独特型活性。我们尝试了几种细胞融合，得到了数千个杂交瘤和一百多个产生具有雌激素样活性的物质的单克隆细胞。进一步分析了这些物质的生物学、生化和化学特性。体内、外生物学活性测定结果显示该物质具有雌激素样活性，但生化分析结果表明该物质不是免疫球蛋白，而是低分子量物质。通过高效液相色谱和质谱分析，我们最终确定该物质为雌激素本身。这些发现是非常令人兴奋的，高比例的杂交瘤细胞具有体外合成雌激素的能力。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