Fujino A, Ishibashi A, Koshiba K, Hoshiai O, Aiso S, Yasuda K, Sogo T
Department of Urology, School of Medicine, Kitasato University, Kanagawa, Japan.
Nihon Hinyokika Gakkai Zasshi. 1990 Jan;81(1):37-44. doi: 10.5980/jpnjurol1989.81.37.
Gamma seminoprotein (gamma Sm), a glycoprotein isolated from human seminal plasma with a molecular weight of 29,000 and possibly a serine protease, has been demonstrated to be one of the prostate organ-specific antigens. We established a murine monoclonal antibody (MoAb) to gamma-Sm in order to prove the presence and localization of this protein in the prostate. The hybrid clones were obtained by fusing mouse SP2/O-Ag-14 myeloma cells with splenocytes from Balb/c mouse immunized with the major fractions of gamma-Sm. The enzyme-linked immunosorbent assay was done for antibody screening. After cloning twice in soft agarose, the stable clone, termed 43-21-1-1, was finally chosen. This MoAb, IgG1(kappa), recognized gamma-Sm specifically, which was verified by an immunoblotting assay. The specificity of the MoAb was further evaluated by immunohistochemical study by the avidin biotin complex method. Periodate-lysine-paraformaldehyde-fixed surgical specimens, including the prostate associated with fibromuscular hyperplasia, seminal vesicles, bladder, testis and epididymis, were examined. Formaldehyde (10%)-fixed surgical specimens from patients with adenocarcinoma of the prostate and primary transitional cell carcinoma arising from the periurethral prostatic ducts were also examined. Positive reactions of gamma-Sm were recognized only in the cytoplasm of prostatic glandular epithelial cells and along the luminal surface. Fibrous and muscular tissues always given negative staining. Neither nonprostatic tissues nor transitional cell carcinoma of the prostate were stained positively for gamma-Sm. These results show that this MoAb (43-21-1-1) is quite specific to gamma-Sm and may be useful for the immunohistochemical study with prostatic tissue.
γ-精浆蛋白（γ-Sm）是一种从人精浆中分离出的糖蛋白，分子量为29,000，可能是一种丝氨酸蛋白酶，已被证明是前列腺器官特异性抗原之一。为了证实这种蛋白在前列腺中的存在及定位，我们制备了抗γ-Sm的鼠单克隆抗体（MoAb）。通过将小鼠SP2/O-Ag-14骨髓瘤细胞与用γ-Sm主要组分免疫的Balb/c小鼠的脾细胞融合，获得了杂交克隆。采用酶联免疫吸附测定法进行抗体筛选。在软琼脂糖中进行两次克隆后，最终选择了稳定的克隆，命名为43-21-1-1。这种MoAb，IgG1（κ），特异性识别γ-Sm，这通过免疫印迹测定得到了验证。通过抗生物素蛋白-生物素复合物法进行免疫组织化学研究，进一步评估了该MoAb的特异性。检查了经高碘酸盐-赖氨酸-多聚甲醛固定的手术标本，包括伴有纤维肌增生的前列腺、精囊、膀胱、睾丸和附睾。还检查了来自前列腺腺癌和起源于尿道周围前列腺导管的原发性移行细胞癌患者的10%甲醛固定手术标本。仅在前列腺腺上皮细胞的细胞质中和沿管腔表面识别出γ-Sm的阳性反应。纤维和肌肉组织总是呈阴性染色。非前列腺组织和前列腺移行细胞癌均未被γ-Sm阳性染色。这些结果表明，这种MoAb（43-21-1-1）对γ-Sm具有高度特异性，可能有助于前列腺组织的免疫组织化学研究。
