White Kimber L, Sheth Christopher M, Peachee Vanessa L
Virginia Commonwealth University, Richmond, Virginia 23298, USA.
J Immunotoxicol. 2007 Apr;4(2):153-8. doi: 10.1080/15476910701337688.
In rodents, the Plaque Assay, T-dependent antibody response to sheep erythrocytes (SRBC), has been reported to be a sensitive and predictive functional immune assay for detecting immunomodulatory compounds. However, various laboratories have chosen to use ELISA-based assays for evaluating the primary immune response in rodents. The ELISA-based assays offer several advantages over the Plaque Assay, which make them attractive for use in immunotoxicological evaluations. Among the most popular antigens used in the ELISA-based assays are SRBC and more recently KLH. While the Plaque Assay and the ELISA-based assays are both capable of evaluating the humoral immune response, they are measuring different endpoints. The Plaque Assay focuses primarily on splenic effects. ELISA-based assays, which use serum from immunized animals, are holistic in nature in that these assays measure effects of antibody production on the spleen, lymph nodes, and bone marrow. Depending on the drug or compound evaluated, different effects and degrees of sensitivity can be seen with the Plaque Assay and ELISA-based assays. One recent finding is that the sensitizing dose of KLH used in the KLH ELISA differentially affects the responses observed in rodents. Even within the same species, different strains of mice and rats produce different magnitudes of responses to the same sensitizing dose. A key component of this discussion focuses on the sensitivity of the Plaque Assay as compared to KLH ELISA-based assays. These assays were evaluated by comparing the response obtained following administration of several known immunosuppressive agents, including cyclophosphamide, azathioprine, cyclosporine A and dexamethasone. The effects on the primary IgM immune response in the B(6)C(3)F(1) mice, the primary immunotoxicological rodents used by National Toxicology Program, and in the Sprague-Dawley rat, the primary rodent models used by industry are addressed.
在啮齿动物中，噬斑测定法，即对绵羊红细胞（SRBC）的T细胞依赖性抗体反应，已被报道是一种用于检测免疫调节化合物的灵敏且具有预测性的功能性免疫测定法。然而，各个实验室已选择使用基于酶联免疫吸附测定（ELISA）的方法来评估啮齿动物的初次免疫反应。与噬斑测定法相比，基于ELISA的方法具有多个优点，这使得它们在免疫毒理学评估中颇具吸引力。基于ELISA的方法中最常用的抗原是SRBC，以及最近使用的钥孔血蓝蛋白（KLH）。虽然噬斑测定法和基于ELISA的方法都能够评估体液免疫反应，但它们测量的是不同的终点。噬斑测定法主要关注脾脏效应。基于ELISA的方法使用免疫动物的血清，本质上是整体性的，因为这些方法测量抗体产生对脾脏、淋巴结和骨髓的影响。根据所评估的药物或化合物不同，噬斑测定法和基于ELISA的方法会观察到不同的效应和敏感度。最近的一项发现是，KLH ELISA中使用的KLH致敏剂量对啮齿动物中观察到的反应有不同影响。即使在同一物种内，不同品系的小鼠和大鼠对相同致敏剂量的反应程度也不同。本次讨论的一个关键部分聚焦于噬斑测定法与基于KLH ELISA的方法相比的敏感度。通过比较给予几种已知免疫抑制剂（包括环磷酰胺、硫唑嘌呤、环孢素A和地塞米松）后获得的反应，对这些测定法进行了评估。文中探讨了对国家毒理学计划使用的主要免疫毒理学啮齿动物B(6)C(3)F(1)小鼠以及业界使用的主要啮齿动物模型斯普拉格 - 道利大鼠的初次IgM免疫反应的影响。
