Schlueter A J, Segre D, Kuhlenschmidt M S, Segre M
Department of Veterinary Pathobiology, University of Illinois, Urbana 61801.
Cell Immunol. 1992 Oct 15;144(2):311-23. doi: 10.1016/0008-8749(92)90247-m.
A minimal requirement in investigations of the behavior of the idiotypic network during immunization is the ability to quantitate both the idiotypic (Ab1) and anti-idiotypic (Ab2) responses. Quantitation of Ab2 in serum is complicated by the simultaneous presence of Ab1, so that Ab1-Ab2 immune complexes escape detection. In contrast, immune complexes should not complicate the enumeration of Ab2-producing lymphocytes in a hemolytic plaque assay. This study utilizes a procedure that allows detection of Ab2-producing cells in such an assay. The procedure relies upon the insertion of the appropriate antibody (Ab1) into the membrane of indicator SRBC through a covalently attached dipalmitoyl phosphatidylethanolamine (DPPE) tail. When the Ab2 response following murine immunization with DNP-Ficoll was analyzed using such an assay, peak plaque-forming cell (PFC) numbers were found to coincide with peak Ab1 PFC numbers in both the primary and secondary response. In addition, this Ab2 response was found to be T independent. The murine immune response to DNP-HGG demonstrated a peak Ab2 PFC response which followed the peak Ab1 PFC response after both primary and secondary immunization. This Ab2 response appeared to be T dependent. The secondary responses to both DNP-Ficoll and DNP-HGG showed increased levels of Ab2 PFC and decreased levels of Ab1 PFC in comparison to the primary responses to the same antigens, suggesting that immunoregulation may occur within these idiotypic networks.
在免疫过程中对独特型网络行为进行研究的一个最低要求是能够对独特型(Ab1)和抗独特型(Ab2)反应进行定量。血清中Ab2的定量因Ab1的同时存在而变得复杂,以至于Ab1-Ab2免疫复合物无法被检测到。相比之下,免疫复合物不应使溶血空斑试验中产生Ab2的淋巴细胞计数变得复杂。本研究采用了一种在这种试验中能够检测产生Ab2细胞的方法。该方法依赖于通过共价连接的二棕榈酰磷脂酰乙醇胺(DPPE)尾将适当的抗体(Ab1)插入指示性绵羊红细胞(SRBC)的膜中。当使用这种试验分析用二硝基苯-聚蔗糖(DNP-Ficoll)免疫小鼠后的Ab2反应时,发现在初次和二次反应中,空斑形成细胞(PFC)的峰值数量与Ab1 PFC的峰值数量一致。此外,发现这种Ab2反应是T细胞非依赖性的。小鼠对二硝基苯-人γ球蛋白(DNP-HGG)的免疫反应在初次和二次免疫后均表现出Ab2 PFC反应峰值,该峰值出现在Ab1 PFC反应峰值之后。这种Ab2反应似乎是T细胞依赖性的。与对相同抗原的初次反应相比,对DNP-Ficoll和DNP-HGG的二次反应均显示Ab2 PFC水平升高,Ab1 PFC水平降低,这表明在这些独特型网络中可能发生免疫调节。