Van Loveren H, Verlaan A P, Vos J G
National Institute of Public Health and Environmental Protection, Bilthoven, The Netherlands.
Int J Immunopharmacol. 1991;13(6):689-95. doi: 10.1016/0192-0561(91)90181-6.
In order to investigate the immune system as a possible target for toxicity by chemicals, functional assays of the immune system are required. Sheep red blood cells (SRBC) are widely used as an antigen to study the immune competence of animals. Since routine toxicology includes data mainly provided with rat toxicity studies, there is a need for easy, specific, and reproducible assays of antibody responses to sheep erythrocytes in the rat. This report describes an enzyme-linked immunosorbent assay (ELISA) of anti-SRBC antibodies of the classes M, G and A in the serum of rats. Antigen preparations, prepared from ghosts of sheep erythrocytes by extraction with KCl, were used to coat the bottom of wells in 96 well microtiter plates. Serum samples from rats, that were immunized with sheep erythrocytes, were titrated on these plates, using specific polyclonal antibodies to rat immunoglobulins M and G, to which peroxidase was conjugated. Immunoglobulin A was assayed, using monoclonal anti-rat IgA antibodies, and polyclonal rat anti-mouse IgG that was conjugated with peroxidase. The results indicate that the ELISA of serum titers of IgM, G and A to sheep erythrocytes is an easy and reliable method, that can be used to detect the effects of chemicals on the immune system of the rat.
为了研究免疫系统作为化学物质毒性的一个可能靶点，需要对免疫系统进行功能检测。绵羊红细胞（SRBC）被广泛用作抗原以研究动物的免疫能力。由于常规毒理学主要包括大鼠毒性研究提供的数据，因此需要一种简单、特异且可重复的方法来检测大鼠对绵羊红细胞抗体反应。本报告描述了一种检测大鼠血清中M、G和A类抗SRBC抗体的酶联免疫吸附测定（ELISA）。用KCl提取绵羊红细胞的细胞膜制备抗原制剂，用于包被96孔微量滴定板的孔底。用绵羊红细胞免疫的大鼠血清样本在这些板上进行滴定，使用与过氧化物酶偶联的抗大鼠免疫球蛋白M和G的特异性多克隆抗体。使用单克隆抗大鼠IgA抗体和与过氧化物酶偶联的多克隆大鼠抗小鼠IgG检测免疫球蛋白A。结果表明，检测血清中IgM、G和A对绵羊红细胞的滴度的ELISA是一种简单可靠的方法，可用于检测化学物质对大鼠免疫系统的影响。
