从人恶性T细胞中纯化出两种抑制因子并进行生化特性分析。

Purification to homogeneity and biochemical characterization of two suppressor factors from human malignant T-cells.

作者信息

Ponzoni M, Montaldo P G, Lanciotti M, Castagnola E, Cirillo C, Cornaglia-Ferraris P

机构信息

Pediatric Oncology Research Laboratory, G. Gaslini Children's Hospital, Italy.

出版信息

Biochem Biophys Res Commun. 1988 Jan 29;150(2):702-10. doi: 10.1016/0006-291x(88)90448-2.

DOI:10.1016/0006-291x(88)90448-2

PMID:3257694

Abstract

A cell clone (GI-CO-T-9) derived from a long term T-cell culture (PF-382), established from a patient affected by acute T-lymphoblastic leukemia (T-ALL), was selected for the presence in the culture medium of factors suppressing T-cell proliferation. The crude supernatant has been subjected to a multi-step chromatographic fractioning, including: preparative gel permeation, anion exchange, and hydrophobic interaction High Performance Liquid Chromatography (HPLC). The highly purified material was characterized by polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE), revealing single bands of 115 Kd and 80 Kd. The isoelectric points (pI), determined by flat-bed isoelectric-focusing, were 7.4 for High Molecular Weight Suppressor Factor (HMWSF) and 3.5-3.6 for Low Molecular Weight Suppressor Factor (LMWSF).

摘要

从一名急性T淋巴细胞白血病（T-ALL）患者建立的长期T细胞培养物（PF-382）中获得的细胞克隆（GI-CO-T-9），因其培养基中存在抑制T细胞增殖的因子而被选中。粗提上清液经过多步色谱分级分离，包括：制备性凝胶渗透、阴离子交换和疏水相互作用高效液相色谱（HPLC）。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）对高度纯化的物质进行表征，显示出115 Kd和80 Kd的单一条带。通过平板等电聚焦测定的等电点（pI），高分子量抑制因子（HMWSF）为7.4，低分子量抑制因子（LMWSF）为3.5 - 3.6。