The purification and properties of human T cell growth factor.
The purification and properties of human T cell growth factor.
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人T细胞生长因子的纯化及性质。
DOI：
10.4049/jimmunol.128.3.1122
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发表时间：
1982
影响因子：
4.4
通讯作者：
R. Gallo
中科院分区：
文献类型：
--
作者：
J. Mier;R. Gallo
用DEAE-Sepharose离子交换层析、Ultrogel AcA 54凝胶过滤和制备型SDS-聚丙烯酰胺凝胶电泳，从无血清淋巴细胞条件培养液中纯化了人T细胞生长因子（TCGF）800倍以上。这种促有丝分裂蛋白在暴露于凝集素（PHA）后由T细胞释放到孵育培养基中，具有m.w.通过SDS-PAGE测定为13，000，通过凝胶过滤测定为20，000至25，000，并且具有6.8的等电点。从丙烯酰胺凝胶中提取的物质在分析SDS凝胶上显示为单一条带（或两条几乎重叠的条带）。部分纯化的材料即使在-70 ° C下也不稳定，并且需要添加牛血清白蛋白或聚乙二醇以保持生物活性。它对蛋白水解消化敏感，但对核酸酶和巯基还原剂如二硫苏糖醇有抗性。人TCGF可用尿素或SDS可逆变性。从丙烯酰胺凝胶中提取的材料已被证明在组织培养中维持T淋巴母细胞。与凝集素、某些抗原和粗淋巴细胞条件培养基相反，纯化的TCGF不启动淋巴细胞胚细胞生成，但对先前通过暴露于凝集素或抗原而活化的T细胞是高度选择性的有丝分裂原。
Human T cell growth factor (TCGF) has been purified more than 800-fold from serum-free lymphocyte conditioned media by utilizing ion exchange chromatography with DEAE-Sepharose, gel filtration with Ultrogel AcA54, and preparative SDS-polyacrylamide gel electrophoresis. This mitogenic protein, which is released by T cells into the incubating media after exposure to a lectin (PHA), has a m.w. of 13,000 as determined by SDS-PAGE and 20,000 to 25,000 on gel filtration and has an isoelectric point of 6.8. The material extracted from acrylamide gels is a single band (or two nearly superimposed bands) when rerun on analytical SDS gels. Partially purified material is unstable even at -70 degrees C and requires the addition of bovine serum albumin or polyethylene glycol to maintain biologic activity. It is sensitive to proteolytic digestion but resistant to nucleases and thiol-reducing agents such as dithiothreitol. Human TCGF can be reversibly denatured with urea or SDS. Material extracted from acrylamide gels has been shown to sustain T lymphoblasts in tissue culture. In contrast to lectins, certain antigens, and crude lymphocyte conditioned media, purified TCGF does not initiate lymphocyte blastogenesis but is a highly selective mitogen for T cells previously activated by exposure to lectins or antigens.