Temple L, Kawabata T T, Munson A E, White K L
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond 23298.
Fundam Appl Toxicol. 1993 Nov;21(4):412-9. doi: 10.1006/faat.1993.1116.
The humoral immune response against sheep red blood cells (SRBC) is one of the most sensitive and frequently used end-points in evaluating the immunotoxicity of drugs and chemicals in experimental animals. Currently, most immunotoxicology studies measure the SRBC IgM antibody response by quantitating the number of SRBC-specific IgM antibody-forming cells using the hemolytic plaque assay. On the other hand, measurement of serum SRBC-specific IgM could be an easier, more cost effective endpoint in evaluating the SRBC antibody response in rodents. A validated method to measure SRBC-specific IgM, however, has not been developed. Thus, the objectives of the studies presented were to develop and validate an enzyme-linked immunosorbent assay (ELISA) for SRBC-specific IgM. Hemoglobin-free, detergent-solubilized membrane preparations were chosen as antigen for the ELISA. Various sources of SRBC were found to be equally useful, and as little as 0.1 micrograms of protein per well was optimal. Kinetic studies of the IgM response showed the peak day to be on Day 5 (mice) and Day 6 (rats). To validate the usefulness of the method for immunotoxicologic studies, serum SRBC-specific IgM levels and number of SRBC-specific plaque-forming cells were compared in rats and mice treated with two well-characterized immunosuppressive agents: benzo[a]pyrene and cyclophosphamide. Administration of these chemicals was found to produce very similar dose-dependent decreases in serum SRBC IgM and IgM-specific plaque-forming cells. These two endpoints were equally sensitive to the effects of the immunosuppressive drugs.(ABSTRACT TRUNCATED AT 250 WORDS)
针对绵羊红细胞（SRBC）的体液免疫反应是评估实验动物中药物和化学物质免疫毒性时最敏感且常用的终点指标之一。目前，大多数免疫毒理学研究通过溶血空斑试验定量SRBC特异性IgM抗体形成细胞的数量来测量SRBC IgM抗体反应。另一方面，测量血清SRBC特异性IgM可能是评估啮齿动物SRBC抗体反应的一种更简便、成本效益更高的终点指标。然而，尚未开发出一种经过验证的测量SRBC特异性IgM的方法。因此，本文所述研究的目的是开发并验证一种用于SRBC特异性IgM的酶联免疫吸附测定（ELISA）方法。选择无血红蛋白、经去污剂溶解的膜制剂作为ELISA的抗原。发现各种来源的SRBC同样有用，每孔低至0.1微克蛋白质最为适宜。IgM反应的动力学研究表明，峰值出现在第5天（小鼠）和第6天（大鼠）。为验证该方法在免疫毒理学研究中的实用性，在分别用两种特征明确的免疫抑制剂：苯并[a]芘和环磷酰胺处理的大鼠和小鼠中，比较了血清SRBC特异性IgM水平和SRBC特异性空斑形成细胞数量。发现给予这些化学物质会导致血清SRBC IgM和IgM特异性空斑形成细胞出现非常相似的剂量依赖性降低。这两个终点指标对免疫抑制药物的作用同样敏感。（摘要截选至250词）
