Karch H, Leying H, Goroncy-Bermes P, Kroll H P, Opferkuch W
Infect Immun. 1985 Nov;50(2):517-22. doi: 10.1128/iai.50.2.517-522.1985.
We recently described how a fraction of isolated fimbriae from a multifimbriated strain of Escherichia coli O7:K1:H6 (WF96) could be subdivided by sequential disaggregation in disrupting agents into individual subunits with different molecular weights. In this study, antibodies were raised in rabbits against these isolated fimbrial subunits and against purified intact WF96 fimbriae. These sera were tested by Western blot analysis or by enzyme-linked immunosorbent assays for reactivity against the following antigens: intact WF96 fimbriae, dissociated WF96 fimbriae, dissociated and reaggregated WF96 fimbriae, the WF96 21K fimbrial subunit, reaggregated WF96 21K subunits, the WF96 16K subunits, reaggregated WF96 16K subunits, intact fimbriae from four other E. coli strains, and deaggregated fimbriae from these strains. We found that antibody against intact WF96 fimbriae only reacted strongly with intact WF96 fimbriae, depolymerized and reaggregated WF96 fimbriae, or reaggregated fimbrial subunits; no reactions were evident with intact fimbriae from four other E. coli strains. Conversely, antisera prepared against the WF96 16K subunit and against the WF96 21K subunit did not react with intact WF96 fimbriae or with depolymerized and reaggregated WF96 fimbriae, but did react with homologous isolated subunits. One cross-reaction between fimbrial subunits was apparent: anti-WF96 16K subunit bound to a 21K subunit of deaggregated fimbriae, from another E. coli strain. Taken together, the findings indicate that the three-dimensional structure of the fimbrial preparation used to immunize animals determines the specificity of the immune response.
我们最近描述了如何通过在裂解剂中进行顺序解离，将来自多菌毛大肠杆菌O7:K1:H6（WF96）菌株的一部分分离菌毛细分为具有不同分子量的单个亚基。在本研究中，用兔制备了针对这些分离的菌毛亚基和纯化的完整WF96菌毛的抗体。通过蛋白质印迹分析或酶联免疫吸附测定法检测这些血清对以下抗原的反应性：完整的WF96菌毛、解离的WF96菌毛、解离并重新聚集的WF96菌毛、WF96 21K菌毛亚基、重新聚集的WF96 21K亚基、WF96 16K亚基、重新聚集的WF96 16K亚基、来自其他四种大肠杆菌菌株的完整菌毛，以及来自这些菌株的解聚菌毛。我们发现，针对完整WF96菌毛的抗体仅与完整的WF96菌毛、解聚并重新聚集的WF96菌毛或重新聚集的菌毛亚基发生强烈反应；与其他四种大肠杆菌菌株的完整菌毛没有明显反应。相反，针对WF96 16K亚基和WF96 21K亚基制备的抗血清不与完整的WF96菌毛或解聚并重新聚集的WF96菌毛反应，但与同源的分离亚基反应。菌毛亚基之间存在一种交叉反应：抗WF96 16K亚基与来自另一种大肠杆菌菌株的解聚菌毛的21K亚基结合。综上所述，这些发现表明用于免疫动物的菌毛制剂的三维结构决定了免疫反应的特异性。
