Development of novel humanized NOG mice producing human IgG antibody

开发产生人 IgG 抗体的新型人源化 NOG 小鼠

• 批准号: 19700373
• 负责人: ITO Ryoji
• 金额: $ 1.98万
• 依托单位: Central Institute for Experimental Animals
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2007
• 资助国家: 日本
• 起止时间: 2007 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-19700373/
• 关键词: ヒト化マウス; ヒト型抗体; B2細胞

The purpose in this research is to produce human IgG antibody using humanized NOG mice that were reconstituted with the human immune system. In this study, we attempted to develop five strains of transgenic (Tg) NOG mice carrying human genes (hIL-7, hLymphotoxin, hCXCL13, hIL-6 and hIFNγ), which are important for the development and functional maturation of immune cells. We confirmed the expression of transgenes, hIL-7, hLymphotoxin and hIL-6 in three strains of Tg mice. Hematopoietic stem cell transplantation into hLymphotoxin Tg mice and substitution of the genetic background of hIL-6 Tg mice in NOG mice are currently underway. We report here that the results obtained by hIL-7 Tg NOG mice reconstituted with the human immune system. We analyzed engraftment and differentiation of human T and B cells in hIL-7 Tg mice by flow cytometry. As a result, the engraftment rate of human cells and differentiation into T and B cells did not differ between hIL-7 Tg NOG and control non-Tg NOG mice. To determine the potential of cell growth of human T cells developed in hIL-7 Tg NOG mice, we isolated human T cells from the spleen and stimulated them with anti-human CD3 and CD28 antibodies in vitro. As a result, human T cells from hIL-7 Tg NOG mice showed significantly higher proliferation than those from non-Tg NOG mice. This result suggests that IL-7 signaling can induce the growth phase, in differentiated human T cells, which presumably become capable of rapid elimination of foreign pathogens. Therefore, hIL-7 Tg NOG mice have potential as a useful tool in the form of humanized mice reconstituted with the human immune system. We are attempting to produce human IgG antibodies by immunization of T cell dependent antigens using hIL-7 Tg NOG mice.

本研究的目的是使用人源化NOG小鼠，用人免疫系统重建，生产人IgG抗体。在本研究中，我们试图开发五种转基因（Tg）NOG小鼠携带人类基因（hIL-7，hIL-6，hCXCL 13，hCXCL 13和hIFNγ），这是重要的免疫细胞的发育和功能成熟。我们证实了转基因hIL-7、hIL-6和hIL-6在三个品系的Tg小鼠中的表达。目前正在进行造血干细胞移植到hIL-6光敏素Tg小鼠中以及在NOG小鼠中替换hIL-6 Tg小鼠的遗传背景。我们在这里报告的hIL-7 Tg NOG小鼠与人类免疫系统重建所获得的结果。我们通过流式细胞术分析了人T和B细胞在hIL-7 Tg小鼠中的植入和分化。结果，在hIL-7 Tg NOG和对照非Tg NOG小鼠之间，人细胞的植入率和向T和B细胞的分化没有差异。为了确定在hIL-7 Tg NOG小鼠中发育的人T细胞的细胞生长潜力，我们从脾分离人T细胞，并在体外用抗人CD 3和CD 28抗体刺激它们。结果，来自hIL-7 Tg NOG小鼠的人T细胞比来自非Tg NOG小鼠的人T细胞显示出显著更高的增殖。这一结果表明，IL-7信号传导可以诱导分化的人T细胞的生长期，这可能成为能够快速消除外来病原体。因此，hIL-7 Tg NOG小鼠具有作为用人免疫系统重建的人源化小鼠形式的有用工具的潜力。我们正试图通过使用hIL-7 Tg NOG小鼠免疫T细胞依赖性抗原来产生人IgG抗体。

项目成果

IFNγ producing killer dendritic cells have a suppressive effect on xeno-GVHD in NOG mice

产生 IFNγ 的杀伤树突状细胞对 NOG 小鼠异种 GVHD 有抑制作用

• 发表时间: 2009
• 作者: Ryoji Ito;Ikumi Katano;Kenji Kawai;Mamoru Ito
• 通讯作者: Mamoru Ito

ヒト Delta-like 1トランスジェニックNOGマウスに発症する骨硬化はヒト造血系の再構築を抑制する

人类 Delta-like 1 转基因 NOG 小鼠中发生的骨硬化抑制人类造血系统的重塑

• 发表时间: 2008
• 作者: Matsuda;T.;Yabushita;H.;Kanaly;R.A.;Shibutani;S.;Yokoyama;A.;伊藤亮治
• 通讯作者: 伊藤亮治

Dendritic cells have a suppressive effect on xeno-GVHD in novel immunodeficient NOG mice

树突状细胞对新型免疫缺陷NOG小鼠中的异种GVHD具有抑制作用

• 发表时间: 2008
• 作者: Ryoji ITO;Ikumi KATANO;Tomoyuki OGURA;Kenji KAWAI;Mamoru ITO
• 通讯作者: Mamoru ITO

Humanized Mice Current Topics in Microbiology and Immunology

人源化小鼠微生物学和免疫学的当前主题

• 发表时间: 2008
• 作者: Cheng XW;Kuzuya M;Nakamura K;Maeda K;Tsuzuki M;Kim W;Sasaki T;Liu Z;Inoue N;Kondo T;Jin H;Numaguchi Y;Okumura K;Yokota M;Iguchi A;Murohara T;R. Ito
• 通讯作者: R. Ito

High sensitivity to xeno-GVHD in severely immunodeficient NOG mice is attributable to dysfunction of dendritic cells

严重免疫缺陷的 NOG 小鼠对异种 GVHD 的高敏感性归因于树突状细胞的功能障碍

• 发表时间: 2009
• 作者: Cheng XW;Murohara T;Kuzuya M;Izawa H;Sasaki T;Obata K;Nagata K;Nishizawa T;Kobayashi M;Yamada T;Kim W;Sato K;Shi GP;Okumura K;Yokota M.;山口智之;伊藤亮治
• 通讯作者: 伊藤亮治