Stackpole C W, De Milio L T, Hämmerling U, Jacobson J B, Lardis M P
Proc Natl Acad Sci U S A. 1974 Mar;71(3):932-6. doi: 10.1073/pnas.71.3.932.
Redistribution of surface immunoglobulins, H-2(b), Thy-1.2, and TL.1,2,3 alloantigens, and concanavalin A receptors on mouse lymphoid cells induced by hybrid rabbit F(ab')(2) antibody (anti-mouse immunoglobulin/anti-visual marker or anti-concanavalin A/anti-visual marker) was studied by immunofluorescence. When used directly to label surface immunoglobulin, and indirectly to label alloantigens and concanavalin A receptors, hybrid antibodies induced similar displacement of all surface components from a uniform distribution into "patches" and "caps" at 37 degrees . One hybrid antibody preparation, antimouse immunoglobulin/anti-ferritin, contained negligible amounts of bivalent anti-mouse immunoglobulin antibody, and was therefore "monovalent" for the antimouse immunoglobulin specificity. This observation suggests that factors other than multivalent crosslinking are responsible for hybrid antibody-induced redistribution of cell-surface components. Cap formation induced by hybrid antibody was enhanced markedly by attachment of the visual marker, either ferritin or southern bean mosaic virus, at 37 degrees . At -5 degrees , hybrid antibody does not displace uniformly distributed H-2(b) alloantigen-alloantibody complexes, but patches of label develop when ferritin attaches to the hybrid antibody. These results explain the patchy distribution of cell-surface components, which is a temperature-independent characteristic of labeling with hybrid antibodies and visual markers for electron microscopy.
通过免疫荧光研究了杂交兔F(ab')(2)抗体（抗小鼠免疫球蛋白/抗可视标记物或抗伴刀豆球蛋白A/抗可视标记物）诱导的小鼠淋巴细胞表面免疫球蛋白、H-2(b)、Thy-1.2以及TL.1、2、3同种异体抗原和伴刀豆球蛋白A受体的重新分布。当直接用于标记表面免疫球蛋白，间接用于标记同种异体抗原和伴刀豆球蛋白A受体时，杂交抗体在37℃诱导所有表面成分从均匀分布类似地移位到“斑块”和“帽”中。一种杂交抗体制剂，抗小鼠免疫球蛋白/抗铁蛋白，含有可忽略量的二价抗小鼠免疫球蛋白抗体，因此对于抗小鼠免疫球蛋白特异性是“单价的”。该观察结果表明，除多价交联外的其他因素是杂交抗体诱导的细胞表面成分重新分布的原因。在37℃时，可视标记物（铁蛋白或南方菜豆花叶病毒）的附着显著增强了杂交抗体诱导的帽形成。在-5℃时，杂交抗体不会使均匀分布的H-2(b)同种异体抗原-同种异体抗体复合物移位，但当铁蛋白附着于杂交抗体时会形成标记斑块。这些结果解释了细胞表面成分的斑块状分布，这是杂交抗体和用于电子显微镜的可视标记物标记的与温度无关的特征。
