Yokota T, Takahashi M, Ishihara T, Yamashita Y, Gondo T, Kawamura S, Hoshii Y, Koga M, Iwata T, Uchino F
First Department of Pathology, Yamaguchi University School of Medicine, Japan.
Acta Pathol Jpn. 1991 Nov;41(11):798-805. doi: 10.1111/j.1440-1827.1991.tb01621.x.
Neutrophils and spleens were prepared from mice after treatment to induce amyloid deposition. The deposition of amyloid was accelerated in normal recipients by intravenous injection of more than 2 x 10(4) neutrophils, and intraperitoneal injection of supernatants obtained by homogenization and centrifugation of the neutrophils and spleens. The supernatants were subjected individually to DEAE ion-exchange chromatography. Amyloid enhancing factor (AEF) activity was present in peaks eluted at an NaCl concentration of 0.17 M. The fractions containing AEF were subjected to high-performance liquid chromatography (HPLC), and AEF was eluted at a position corresponding to about 15 KDa. Purified AEF was analyzed by amino acid sequencing and gas chromatography. The N-terminal amino acid was blocked, and AEF contained some saccharides including glucose, mannose, galactosamine and sialic acid, and an undefined substance (probably derived from certain proteins). Immunoelectron microscopy by the pre-embedding method using an anti-AEF antiserum demonstrated that the cytosol, but not primary and specific granules in neutrophils from the spleen of amyloid-laden mice, reacted with the antiserum. These findings suggest that AEF is a glycoprotein associated with neutrophils.
在诱导淀粉样蛋白沉积的处理后，从小鼠制备中性粒细胞和脾脏。通过静脉注射超过2×10⁴个中性粒细胞，以及腹腔注射通过对中性粒细胞和脾脏进行匀浆和离心获得的上清液，可加速正常受体中淀粉样蛋白的沉积。将上清液分别进行DEAE离子交换色谱分析。在NaCl浓度为0.17 M时洗脱的峰中存在淀粉样增强因子（AEF）活性。将含有AEF的级分进行高效液相色谱（HPLC）分析，AEF在对应于约15 kDa的位置洗脱。通过氨基酸测序和气相色谱对纯化的AEF进行分析。N端氨基酸被封闭，AEF含有一些糖类，包括葡萄糖、甘露糖、半乳糖胺和唾液酸，以及一种未定义的物质（可能源自某些蛋白质）。使用抗AEF抗血清通过包埋前方法进行的免疫电子显微镜显示，来自淀粉样蛋白负荷小鼠脾脏的中性粒细胞中的胞质溶胶，而非初级和特异性颗粒，与抗血清发生反应。这些发现表明AEF是一种与中性粒细胞相关的糖蛋白。
