van de Rijn I, Zabriskie J B, McCarty M
J Exp Med. 1977 Aug 1;146(2):579-99. doi: 10.1084/jem.146.2.579.
Heart-reactive antibody (HRA) appears in the sera of experimental animals inoculated with group A streptococci as well as patients with acute rheumatic fever. Adsorption of either serum with group A streptococcal membranes will remove the HRA. Blocking experiments between these two types of HRAs have demonstrated that the antibodies are directed towards different antigenic determinants on either the same or different molecules. To isolate and purify the antigen from the group A streptococcus cross-reactive with sarcolemmal sheaths of cardiac myofibers, it became necessary to purify the HRA from rheumatic fever patients' sera. Isolated gamma globulin containing all of the HRA was adsorbed onto human sarcolemmal sheaths. The specific HRA was released by using potassium iodide. Over 99 percent of the purified HRA was shown to bind the sarcolemmal sheath whereas less than 1 percent of the antibody would bind nonspecifically to other material. Preparations of group A streptococcal membrane will bind HRA purified from the sera of acute rheumatic patients at levels of 97 percent or greater. The cross-reactive antigen solubilized by nonionic detergent was purified 120-fold by column chromatography. On sodium dodecyl sulfate polyacrylamide electrophoresis, the antigen was demonstrated to be composed of four polypeptides with mol wt of 32,000, 28,000, 26,000, and 22,000 daltons, respectively. Only proteolytic enzymes could destroy the antigenic determinant whereas glycosidases and lipases had no effect. The purified antigen blocked the binding of purified HRA to normal human heart sections.
心脏反应性抗体(HRA)出现在接种A组链球菌的实验动物血清以及急性风湿热患者的血清中。用A组链球菌膜吸附任何一种血清都会去除HRA。这两种类型的HRA之间的阻断实验表明,这些抗体针对的是相同或不同分子上的不同抗原决定簇。为了从与心肌纤维肌膜鞘交叉反应的A组链球菌中分离和纯化抗原,有必要从风湿热患者的血清中纯化HRA。含有所有HRA的分离γ球蛋白被吸附到人类肌膜鞘上。使用碘化钾释放特异性HRA。结果显示,超过99%的纯化HRA与肌膜鞘结合,而不到1%的抗体与其他物质非特异性结合。A组链球菌膜制剂将以97%或更高的水平结合从急性风湿患者血清中纯化的HRA。通过柱色谱法将非离子洗涤剂溶解的交叉反应抗原纯化了120倍。在十二烷基硫酸钠聚丙烯酰胺凝胶电泳上,该抗原被证明由四种分子量分别为32000、28000、26000和22000道尔顿的多肽组成。只有蛋白水解酶能破坏抗原决定簇,而糖苷酶和脂肪酶则没有作用。纯化的抗原阻断了纯化的HRA与正常人心脏切片的结合。