Differentiation-induction of antibody producing human B cells from cord blood CD34+ cells in mice for generating monoclonal antibody used in clinical therapy

从小鼠脐带血 CD34 细胞中分化诱导产生抗体的人 B 细胞，以产生用于临床治疗的单克隆抗体

• 批准号: 12557032
• 负责人: HABU Sonoko
• 金额: $ 8.51万
• 依托单位: Tokai University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557032/
• 关键词: human immune system; T cell development; thymic environment; NOG mice; mice; B1 B cell; antiboy production; B cell development; B_1 B Cells; ヒト免疫系再構築; モノクローナル抗体; ヒト臍帯血; オリゴクローン法

Monoclonal antibodies are known to be useful as the clinical therapy from various animal models. However, therapeutic antibodies derived human B cells have not used in the clinical field although human/mouse chimera antibodies are available. In order to obtain monoclonal antibodies produced by human B cells, we tried to reconstitute human immune system in the immuno-deficient mice from CD34+ cells in cord blood (CB) in this study. We obtained the following results. 1) Human CD34+ cells develop into mature T cells with cytokine producing ability in the in vivo RTOC in which mouse thymic epithelial cells are reaggregated with human CD34+cells. 2) NOG mice are useful to reconstitute T cell development in vivo probably because they lack completely NK cell activity as well as T and B cells. 3) In vivo treatment of anti-CD40 antibody induces the increase of antigen specific antibody although IgG class is low. 4) CB CD34+ cells injected into NOG mice mainly develop into CD5+B cells in the spleen. 5) Using chimera anti-erbB-2 antibody and serum from malaria patients, antigen epitopes were identified. These peptides are going to use for immunizing NOG mice receiving DBCD34+ cells.

从不同的动物模型来看，单抗被认为是有用的临床治疗方法。然而，尽管人/鼠嵌合抗体已经存在，但来源于人类B细胞的治疗性抗体尚未在临床上使用。为了获得人B细胞产生的单抗，本研究试图用脐带血中的CD34+细胞来重建免疫缺陷小鼠的免疫系统。我们得到了以下结果。1)在小鼠胸腺上皮细胞与人CD34+细胞重组的体内RTOC中，人CD34+细胞发育为具有细胞因子产生能力的成熟T细胞。2)NOG小鼠可用于体内重建T细胞发育，可能是因为它们完全缺乏NK细胞活性以及T和B细胞。3)体内应用抗CD40抗体后，虽然免疫球蛋白G水平较低，但仍可诱导产生较高的抗原特异性抗体。4)注射入NOG小鼠体内的CB CD34+细胞主要发育为CD5+B细胞。5)用嵌合体抗erB-2抗体和疟疾患者血清鉴定抗原表位。这些多肽将用于免疫接受DBCD34+细胞的NOG小鼠。

项目成果

Shinya Nagafuchi: "Dietary nucleotides can up-regulate antigen-specific Th 1 immune responses and suppress antigen-specific IgE responsesin mice"Int Arch Allergy Immunol. 122. 33-41 (2000)

Shinya Nagafuchi：“膳食核苷酸可以上调小鼠中抗原特异性 Th 1 免疫反应并抑制抗原特异性 IgE 反应”Int Arch AllergyImmunol。

Wataru Ise: "Primary response of naitive CD4+ T cells to amino acid-substited analogs of an antigenic peptide can show distinct activation patterns : Th1-and Th2-type cytokine secretion, and helper activity for antibody production without apparent cytoki

Wataru Ise：“天然 CD4 T 细胞对氨基酸取代的抗原肽类似物的初级反应可以表现出不同的激活模式：Th1 和 Th2 型细胞因子分泌，以及在没有明显细胞因子的情况下产生抗体的辅助活性

佐藤健人, 赤塚明, 垣生園子: "細胞培養工学"ニューサイエンス社. 5 (2000)

Kento Sato、Akira Akatsuka、Sonoko Kaki：《细胞培养工程》新科学出版 5 (2000)。

Takehiko Sato: "Surface molecules essential for positive selection are retained but Interferod in thymic epithelial cells after monolayer culture"Cellular Immunology. 211. 71-79 (2001)

Takehiko Sato：“阳性选择所必需的表面分子被保留，但单层培养后胸腺上皮细胞中的干扰”细胞免疫学。

Makoto Senoo: "Limited effect of chromatin remodeling on D β-to-JB β recombination in CD4^+CD8^+ thymocyte : Implications for a new aspect in the regulation of TOR β gene recombination"International Immunology. 13. 1405-1414 (2001)

Makoto Senoo：“染色质重塑对 CD4^+CD8^+ 胸腺细胞中 D β-to-JB β 重组的有限影响：对 TOR β 基因重组调控新方面的影响”国际免疫学 13. 1405-1414（ 2001）