The mechanism of interleukin-2 metabolism and its application for the cancer therapy

IL-2代谢机制及其在癌症治疗中的应用

• 批准号: 03454327
• 负责人: YASUTOMI Masayuki
• 金额: $ 2.82万
• 依托单位: Kinki University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03454327/
• 关键词: Interleukin-2; IL-2 clearance; Liver metastasis; Liposome; 肝局所免疫; 肝転移; ガラクト-ス基; インターロイキン2; 生体内代謝; 腎臓; サイトゾール; 半減期; インタ-ロイキン2; マウス; サイトゾ-ル

We reported previously that interleukin-2 (IL-2) metabolizing enzyme exists in the kidney cytosol. The kidney cytosol fraction was purified using ammonium sulfate precipitation, iron exchange chromatography and IL-2 affinity gel chromatography for obtaining high yields of purfied enzyme. The assay using IL-2 as a substrate of purified fraction showed enzymatic activity to be 64-fold higher than a crude fraction. The Km value of the purified fraction showed approximately 50muM, which means highly specific against IL-2. The SDS-PAGE of the purified fraction showed three bands, 12.3 kDa, 25.7kDa and 61.1kDa, and the 61.1kDa band shows a significant high enzymatic activity. By the immunization of this fraction to a rabbit, polyclonal antibody to IL-2 degradation enzyme was detected in sera. However, the administration of sera never prolonged the serum half-life of IL-2 in mice model.Another approach for avoiding the short half-life of IL-2, we have tried a chemically modified IL-2 products using liposome. we noticed galactose-receptors existing adundantly in the liver and prepared galactose-containing liposome-IL-2 preparations using various carriers/bases. We infused these preparations in the portal or caudal vein of mice and investigated the accumulation to organs and localized immunological copetence in the liver. As a result, galactose-containing liposome-IL-2 preparations showed the highest accumulation and the highest localized enhancing effect on the antitumor activity of lymphocytes in the liver.

我们曾报道白细胞介素2（IL-2）代谢酶存在于肾脏胞浆中。采用硫酸铵沉淀、铁交换层析和IL-2亲和凝胶层析对肾胞液组分进行纯化，以获得高产量的纯化酶。使用IL-2作为纯化级分的底物的测定显示酶活性比粗级分高64倍。纯化级分的Km值显示约50 μ M，这意味着对IL-2具有高度特异性。SDS-PAGE电泳结果显示，纯化组分有12.3kDa、25.7kDa和61.1kDa三条带，其中61.1kDa的条带显示出明显的高酶活性。用此组分免疫家兔，在血清中检测到抗IL-2降解酶的多克隆抗体。然而，在小鼠模型中，血清给药并没有延长IL-2的血清半衰期。另一种避免IL-2半衰期短的方法是，我们尝试了使用脂质体的化学修饰的IL-2产品。我们注意到半乳糖受体大量存在于肝脏中，并使用各种载体/基质制备含半乳糖的脂质体-IL-2制剂。我们将这些制剂输注到小鼠的门静脉或尾静脉中，并研究了在器官中的蓄积和在肝脏中的局部免疫功能。结果，含半乳糖的脂质体-IL-2制剂显示出最高的蓄积和对肝脏中淋巴细胞的抗肿瘤活性的最高局部增强作用。

项目成果

Okuno,K.,et al.: "A successful liver metastasis model in mice with neuraminidase treated colon 26." Surgery Today. 23. 795-799 (1993)

Okuno,K.,et al.：“用神经氨酸酶处理结肠 26 的小鼠成功建立了肝转移模型。”

Okuno,K.,et al: "Effect of packed red cell and whole blood transfusion on liver-associated immune function.(in press)" Am.j Surg.(1993)

Okuno,K. 等人：“浓缩红细胞和全血输注对肝脏相关免疫功能的影响。（正在印刷中）”Am.j Surg.(1993)

Ojuno,K.et al.: "Effect of packed red cell and whole blood transfusion on liver-associated immmune function." Am.J.Surg.(in press). (1993)

Ojuno,K.等人：“浓缩红细胞和全血输注对肝脏相关免疫功能的影响。”

Okuno,K.,et al.: "A successful liver metastasis model in mice with neuramindase treated colon 26." Surgeray Today. 23. 795-799 (1993)

Okuno,K.,et al.：“用神经氨酸酶处理结肠 26 的小鼠成功建立了肝转移模型。”

大西 博昭,奥野 清隆,安富 正幸: "インタ-ロイキン2の生体内代謝" 臨床免疫. 24巻4号.

Hiroaki Onishi、Kiyotaka Okuno、Masayuki Yasutomi：“白细胞介素 2 的体内代谢”《临床免疫学》第 24 卷，第 4 期。