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公开(公告)号：US08716216B2
公开(公告)日：2014-05-06
申请号：US12118675
申请日：2008-05-09
CPC分类号： C07K14/4703 , A61K38/00 , Y02A50/471
摘要： A composition of an immunosuppressant protein (HISP) which is achieved by the steps of obtaining supernatant from hNT neuronal cells; exposing the supernatant to preparative polyacrylamide gel; placing the active isoelectric fraction on a Blue Sepharose column to bind albumin; and collecting the free fraction containing the concentrated, isolated 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. 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 act through the T-cell receptor-CD3 complex or via altered accessory signal cells.
摘要翻译： 通过从hNT神经元细胞获得上清液的步骤实现的免疫抑制蛋白（HISP）的组合物; 将上清液暴露于制备型聚丙烯酰胺凝胶; 将活性等电点部分置于蓝色琼脂糖凝胶柱上以结合白蛋白; 并收集含有浓缩，分离的HISP的游离级分。 HISP为阴离子型，分子量为40-100kDa，等电点为约4.8，由hNT细胞的上清液得到。 HISP可以抑制同种异体混合淋巴细胞培养物中应答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），离子霉素和伴刀豆素A的T细胞增殖和IL-2产生。 HISP不通过T细胞受体-CD3复合物或通过改变的附属信号细胞起作用。
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公开(公告)号：US20140234401A1
公开(公告)日：2014-08-21
申请号：US14220714
申请日：2014-03-20
IPC分类号： C07K14/47
CPC分类号： C07K14/4703 , A61K38/00 , Y02A50/471
摘要： A method of treating inflammation by administering a therapeutically effective amount of a human immunosuppressant protein (HISP) to a subject is presented. The inventors have discovered a novel immunosuppressive protein purified from the supernatant of hNT cell culture. The immunosuppressant protein has a molecular weight of about 40-100 kDa, an isoelectric point of about 4.4, a net ionic charge and is capable of suppressing T-cell activation, T-cell proliferation and the production of IL-2. This protein can be used in treating inflammation, preventing graft rejection after transplantation, treating autoimmune diseases and suppressing allergic responses as well as other uses.
摘要翻译： 提出了通过向受试者施用治疗有效量的人免疫抑制蛋白（HISP）来治疗炎症的方法。 本发明人发现了从hNT细胞培养物的上清液中纯化的新型免疫抑制蛋白。 免疫抑制蛋白具有约40-100kDa的分子量，约4.4的等电点，净离子电荷，并且能够抑制T细胞活化，T细胞增殖和IL-2的产生。 该蛋白质可用于治疗炎症，预防移植后的移植排斥反应，治疗自身免疫疾病和抑制过敏反应以及其他用途。
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公开(公告)号：US09051386B2
公开(公告)日：2015-06-09
申请号：US14220714
申请日：2014-03-20
CPC分类号： C07K14/4703 , A61K38/00 , Y02A50/471
摘要： A method of treating inflammation by administering a therapeutically effective amount of a human immunosuppressant protein (HISP) to a subject is presented. The inventors have discovered a novel immunosuppressive protein purified from the supernatant of hNT cell culture. The immunosuppressant protein has a molecular weight of about 40-100 kDa, an isoelectric point of about 4.4, a net ionic charge and is capable of suppressing T-cell activation, T-cell proliferation and the production of IL-2. This protein can be used in treating inflammation, preventing graft rejection after transplantation, treating autoimmune diseases and suppressing allergic responses as well as other uses.
摘要翻译： 提出了通过向受试者施用治疗有效量的人免疫抑制蛋白（HISP）来治疗炎症的方法。 本发明人发现了从hNT细胞培养物的上清液中纯化的新型免疫抑制蛋白。 免疫抑制蛋白具有约40-100kDa的分子量，约4.4的等电点，净离子电荷，并且能够抑制T细胞活化，T细胞增殖和IL-2的产生。 该蛋白质可用于治疗炎症，预防移植后的移植排斥反应，治疗自身免疫疾病和抑制过敏反应以及其他用途。
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公开(公告)号：US07388076B2
公开(公告)日：2008-06-17
申请号：US10621061
申请日：2003-07-16
CPC分类号： C07K14/4703 , A61K38/00 , Y02A50/471
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-1 monocytic 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： 纯化免疫抑制蛋白（HISP）的方法具有从hNT细胞获得上清液的步骤; 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，包括活性等电点＃10; 将活性等电点部分置于蓝色琼脂糖凝胶柱上以结合白蛋白; 并收集含有浓缩，分离的HISP的游离级分。 还公开了使用有效量的HISP治疗炎症的方法。 HISP是阴离子型的，分子量为40-100kDa，等电点约为4.8，从hNT细胞的上清液获得，而不是来自NCCIT胚胎癌细胞，T98G胶质母细胞瘤细胞或THP-1单核细胞白血病细胞。 HISP可以将T细胞维持在静止状态，而不降低其活力。 当用热，pH2，pH11处理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可以抑制同种异体混合淋巴细胞培养物中应答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），离子霉素和伴刀豆素A的T细胞增殖和IL-2产生。 HISP不结合肝素 - 琼脂糖CL-B凝胶; 或白蛋白结合树脂蓝色琼脂糖。 HISP集中在YM10超滤。 HISP不通过T细胞受体-CD3复合物或通过改变的附属信号细胞起作用。 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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公开(公告)号：US20080280812A1
公开(公告)日：2008-11-13
申请号：US12118675
申请日：2008-05-09
CPC分类号： C07K14/4703 , A61K38/00 , Y02A50/471
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-1 monocytic leukemia cells. HISP can maintain T cells in a quiescent G.sub.0/G.sub.1 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： 纯化免疫抑制蛋白（HISP）的方法具有从hNT细胞获得上清液的步骤; 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，包括活性等电点＃10; 将活性等电点部分置于蓝色琼脂糖凝胶柱上以结合白蛋白; 并收集含有浓缩，分离的HISP的游离级分。 还公开了使用有效量的HISP治疗炎症的方法。 HISP是阴离子型的，分子量为40-100kDa，等电点约为4.8，从hNT细胞的上清中获得，但不是来自NCCIT胚胎癌细胞。 T98G胶质母细胞瘤细胞或THP-1单核细胞白血病细胞。 HISP可以将T细胞维持在静止的G.sub.0 / G.sub.1状态，而不降低其活力。 当用热，pH2，pH11处理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可以抑制同种异体混合淋巴细胞培养物中应答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），离子霉素和伴刀豆素A的T细胞增殖和IL-2产生。 HISP不结合肝素 - 琼脂糖CL-B凝胶; 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附属信号细胞起作用。 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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