Cardillo F, Mengel J, Garcia S B, Cunha F Q
Department of Pharmacology, Faculty of Medicine of Ribeirão Preto, University of São Paulo, Brazil.
J Immunol Methods. 1995 Dec 15;188(1):43-9. doi: 10.1016/0022-1759(95)00200-6.
The production of monoclonal antibodies to protein antigens which can only be obtained in tiny amounts has been a major task, since classical in vivo immunization procedures are not always efficient. In order to circumvent this problem, two methods have been developed: (1) in vitro immunization, in which the immunogen is presented directly to spleen cell cultures; (2) intrasplenic immunization, a technique in which the immunogen is deposited in the spleen tissue. The latter approach requires less laboratory work and the risk of contamination, often a problem with in vitro cultures (Nilsson and Larsson, Immunol. Today (1990) 11, 10), is greatly reduced. Here, we describe a novel method of grafting neonatal spleens in the pinna of the mouse ear. Histological and functional studies show that these spleen grafts have white and red pulp and contain normal percentages of functional T and B cells. The results indicate that this procedure is extremely efficient in priming mice for a secondary humoral immune response, since very small amounts of soluble antigen (ovalbumin) were required. The data are discussed in terms of the advantages of this new technique over current procedures for intrasplenic immunization.
针对只能少量获取的蛋白质抗原生产单克隆抗体一直是一项重大任务，因为传统的体内免疫程序并不总是有效。为了规避这一问题，已开发出两种方法：（1）体外免疫，即将免疫原直接呈递给脾细胞培养物；（2）脾内免疫，一种将免疫原注入脾脏组织的技术。后一种方法所需的实验室工作较少，而且污染风险（体外培养常常存在的问题）（尼尔森和拉尔森，《免疫学今日》（1990年）11卷，10页）大幅降低。在此，我们描述一种将新生脾脏移植到小鼠耳廓的新方法。组织学和功能研究表明，这些脾脏移植物有白髓和红髓，且功能性T细胞和B细胞的比例正常。结果表明，该程序在使小鼠启动二次体液免疫应答方面极其有效，因为只需极少量的可溶性抗原（卵清蛋白）。将根据这项新技术相对于当前脾内免疫程序的优势对这些数据进行讨论。
