Isolation of human antibodies targeting to immunocytes from phage library and regulation of immune response

从噬菌体库中分离针对免疫细胞的人类抗体并调节免疫反应

• 批准号: 13672325
• 负责人: ITO Yuji
• 金额: $ 2.18万
• 依托单位: Kagoshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13672325/
• 关键词: Costimulatory molecules; human antibody; phage library; vaccine; antigen presenting cell; T cell; antagonist; targeting; 免疫応答制御; 単鎖Fv抗体; T細胞増殖; アレルギー; 拒絶反応抑制; 抗体医薬; 遺伝子治療; ワクチン開発; ファージディスプレイ; 阻害抗体; 免疫反応の制御; DNAワクチン

The aims of this study are the isolation of human antibodies directed to the costimulatory molecules on the lymphocytes, the establishment of the targeting and the control of the immune system by using antibodies, and their applications for DNA vaccine development. We succeeded in the isolation of anti B7RP-1 antibodies which could inhibit the costimulatory signal between ICOS and B7RP-1 by human antibody phage display library and are now applying for a patent. The obtained three antibodies could actually repress the T cell proliferation which was induced by anti CD3 antibody stimulation. We also developed other human antibodies that were specific to CD86, a costimulator ligand for CD28 and are now under preparation for a patent. The antibodies mentioned here can function as not only antagonist blocking the costimulatory signal but also targeting molecule to the antigen presenting cells (APC), because APC express abundantly such CD86 and B7RP-1 molecules, and such targeting ability is very important for the effective vaccine development to lead the antigens efficiently to the APC. We are now investigating the new vaccine developing studies by use of our human antibodies. As a preliminary experiment, we constructed a gene of fusion protein of extracellular domain of CTLA-4 (targeting molecule to APC) and human Fc (antigen) and immunized it through skin on mice as DNA vaccine. Consequently high antibody response to the antigen was obtained in the case of the administration of CTLA-4-Fc fusion, as compared with the administration of Fc (antigen) only. These results indicate that the targeting of the antigen to the APC give the promising effect in the vaccine efficacy.

这项研究的目的是通过使用抗体及其对DNA疫苗开发的应用，分离了针对淋巴细胞上淋巴细胞的人类抗体的分离，靶向和免疫系统的控制。我们成功地分离了抗B7RP-1抗体，这些抗体可以通过人类抗体噬菌体显示库抑制ICOS和B7RP-1之间的共刺激信号，现在正在申请专利。获得的三种抗体实际上可以抑制由抗CD3抗体刺激诱导的T细胞增殖。我们还开发了其他针对CD86的人类抗体，CD86是CD28的共可能配体，现在正在准备专利。此处提到的抗体不仅可以用作阻断结肠刺激信号的拮抗剂，而且还可以靶向分子靶向抗原呈递细胞（APC），因为APC表达了这种CD86和B7RP-1分子的大量表达，并且此类靶向能力对于有效的疫苗发育非常重要，可以使抗原有效地对APC有效地引起抗原。我们现在正在研究新的疫苗通过使用人类抗体来开发研究。作为一个初步实验，我们构建了CTLA-4（靶向APC）和人FC（抗原）的细胞外域融合蛋白的基因，并通过小鼠作为DNA疫苗在皮肤上免疫。因此，仅在施用CTLA-4-FC融合中，与施用FC（抗原）相比，获得了对抗原的高抗体反应。这些结果表明，抗原对APC的靶向对疫苗功效产生了有希望的作用。

项目成果

Human Fc ε RI α - Specific Human Single-Chain Fv (scFv) Antibody with Antagonistic Activity toward IgE/Fc ε RI α -Binding

人 Fc ε RI α - 具有 IgE/Fc ε RI α 结合拮抗活性的特异性人单链 Fv (scFv) 抗体

• 发表时间: 2003
• 期刊: J.Biochem. 133
• 作者: S.Hashiguchi;T.Nakashima;A.Nitani;T.Yoshihara;K.Yoshinaga;Y.Ito;Y.Maeda K.Sugimura
• 通讯作者: Y.Maeda K.Sugimura

伊東祐二: "ヒト抗体エンジニアリングと分子標的医療"バイオインダストリー. 20巻7号. 34-42 (2003)

伊藤雄二：“人类抗体工程和分子靶向医学”《生物工业》第 20 卷，第 7. 34-42 期（2003 年）。

橋口周平(他、7名): "Human FcεRIα-Specific Human Single-Chain Fv (scFv) Antibody with Antagonistic Activity to IgE/FcεRIα-Binding"J.Biochem.. 133. 43-49 (2003)

Shuhei Hashiguchi（和其他 7 人）：“对 IgE/FcεRIα-结合具有拮抗活性的人 FcεRIα-特异性人单链 Fv (scFv) 抗体”J.Biochem.. 133. 43-49 (2003)

杉村和久: "ファージディスプレイ法"Molecular Medicine. Vol.40(10). 1150-1158 (2003)

Kazuhisa Sugimura：“噬菌体展示方法”《分子医学》第 40 卷（10）（2003 年）。

伊東祐二: "抗体エンジニアリングの最前線(分担)"シーエムシー出版. 194(9) (2004)

伊藤雄二：《抗体工程前沿（分享）》CMC Publishing 194（9）（2004）。