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1. 发明申请

WO2004007693A3 HUMAN IMMUNOSUPPRESSIVE PROTEIN 审中-公开
标题翻译：人类免疫抑制蛋白
公开(公告)号：WO2004007693A3
公开(公告)日：2004-12-02
申请号：PCT/US0322664
申请日：2003-07-16
申请人： UNIV SOUTH FLORIDA
发明人： SANBERG PAUL R , ENGELMAN ROBERT W , GOWER WILLIAM R
IPC分类号： A61K38/00 , C07K14/47 , C07K1/00 , C07K14/52
CPC分类号： C07K14/4703 , A61K38/00
摘要： A method for purifying an immunosuppressant protein (HISP) has the steps of obtaining supernatant from hNT cells; exposing the supernatant to preparative polyacrylamide gel electrophoresis to produce 20 isoelectric fractions, including active isoelectric fraction #10; placing the active isoelectric fraction on a Blue Sepharose column to bind albumin; and collecting the free fraction containing the concentrated, isolated HISP. Also disclosed is a method of treating inflammation, using an effective amount of an HISP. The HISP is anionic, has a molecular weight of 40-100 kDa, an isoelectric point of about 4.8 and is obtained from the supernatant of hNT cells, but not from NCCIT embryonal carcinoma cells, T98G glioblastoma cells or THP-Imonocytic leukemia cells. HISP can maintain T cells in a quiescent G0/G1 state without lowering their viability. HISP loses activity when treated with heat, pH2, pH11, or mixed with trypsin or carboxypeptidase, but not with neuraminidase. HISP can suppress proliferation of responder peripheral blood mononuclear cells in allogeneic mixed lymphocyte cultures; HISP can suppress T-cell proliferation and IL-2 production in response to phorbol 12-myristate 13-acetate (PMA), ionomycin and concanavalin-A. HISP does not bind to heparin-sepharose CL-B gel; or to albumin-binding resin Blue Sepharose. HISP is concentrated with YM10 ultrafiltration. HISP does not act through the T-cell receptor-CD3 complex or via altered accessory signal cells. A method of treating inflammation comprises administering an effective amount of hNT neuronal cells.
摘要翻译：纯化免疫抑制剂蛋白质（HISP）的方法具有以下步骤：从hNT细胞获得上清液; 将上清液暴露于制备型聚丙烯酰胺凝胶电泳以产生20个等电点部分，包括活性等电点部分＃10; 将活性等电点部分置于Blue Sepharose柱上以结合白蛋白; 并收集含有浓缩的分离HISP的游离部分。还公开了使用有效量的HISP治疗炎症的方法。 HISP是阴离子的，具有40-100kDa的分子量，等电点约为4.8，从hNT细胞的上清液获得，但不是从NCCIT胚胎癌细胞，T98G成胶质细胞瘤细胞或THP-I单核细胞白血病细胞获得。 HISP可将T细胞维持在静止的G0 / G1状态而不降低其活力。当用热，pH2，pH11或与胰蛋白酶或羧肽酶混合时，HISP失去活性，但不与神经氨酸酶混合。 HISP可抑制同种异体混合淋巴细胞培养物中响应者外周血单核细胞的增殖; HISP可以抑制佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA），伊屋诺霉素和伴刀豆球蛋白-A对T细胞增殖和IL-2产生的抑制作用。 HISP不与肝素 - 琼脂糖凝胶CL-B凝胶结合; 或白蛋白结合树脂蓝色琼脂糖。 HISP用YM10超滤浓缩。 HISP不通过T细胞受体-CD3复合物或通过改变的辅助信号细胞起作用。治疗炎症的方法包括施用有效量的hNT神经元细胞。

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