Teuscher C, Wild G C, Tung K S
J Immunol. 1983 Jan;130(1):317-22.
A soluble aspermatogenic autoantigen (AP2) capable of inducing experimental allergic orchitis (EAO) in the guinea pig has been isolated from the soluble acrosomal contents (SAC) released from guinea pig cauda epididymal sperm during the in vitro Ca+2 ionophore A23187-induced acrosome reaction. AP2 purification was achieved by heat inactivation of SAC enzymatic activity, followed by chaotropic solubilization and ultracentrifugation, gel filtration on Sephadex G-50, preparative isoelectric focusing, and SDS-polyacrylamide gel electrophoresis. Approximately 80 micrograms of AP2 was obtained from 1 to 2 x 10(10) cauda epididymal sperm. AP2 has a m.w. of 9500 +/- 1500 as determined by unreduced SDS-PAGE and an isoelectric point of 5.52 +/- 0.11. Amino acid analysis of AP2 indicates that it is a protein. AP2 has no detectable hexose or hexosamine as determined by gas liquid chromatographic analysis and is therefore probably not a glycoprotein. Five micrograms of AP2 are capable of inducing severe EAO in 100% of guinea pigs tested, whereas 0.5 to 2.0 micrograms induces mild lesions consisting primarily of aspermatogenesis in 66% of guinea pigs tested.
一种能够在豚鼠中诱发实验性过敏性睾丸炎（EAO）的可溶性生精自身抗原（AP2）已从豚鼠附睾尾精子在体外钙离子载体A23187诱导的顶体反应过程中释放的可溶性顶体内容物（SAC）中分离出来。通过对SAC酶活性进行热灭活实现AP2的纯化，随后进行离液剂增溶和超速离心、在葡聚糖G - 50上进行凝胶过滤、制备性等电聚焦以及SDS - 聚丙烯酰胺凝胶电泳。从1至2×10（10）个附睾尾精子中获得了约80微克的AP2。通过未还原的SDS - PAGE测定，AP2的分子量为9500±1500，等电点为5.52±0.11。对AP2的氨基酸分析表明它是一种蛋白质。通过气相色谱分析确定，AP2中未检测到己糖或己糖胺，因此它可能不是糖蛋白。5微克的AP2能够在100%的受试豚鼠中诱发严重的EAO，而0.5至2.0微克则在66%的受试豚鼠中诱发主要由无精子症组成的轻度病变。
