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    GENERATION OF NK CELLS AND NK-CELL PROGENITORS
    1.
    发明申请
    GENERATION OF NK CELLS AND NK-CELL PROGENITORS 审中-公开
    NK细胞和NK细胞发生子的产生

    公开(公告)号：WO2012128622A1

    公开(公告)日：2012-09-27

    申请号：PCT/NL2012/050165

    申请日：2012-03-16

    Applicant: IPD-THERAPEUTICS B.V. ,  SPANHOLTZ, Jan

    Inventor： SPANHOLTZ, Jan

    IPC: C12N5/0783

    CPC classification number: C12N5/0646 ,  C12N2501/125 ,  C12N2501/145 ,  C12N2501/21 ,  C12N2501/22 ,  C12N2501/2302 ,  C12N2501/2306 ,  C12N2501/2307 ,  C12N2501/2315 ,  C12N2501/235 ,  C12N2501/26 ,  C12N2501/91

    Abstract: The present invention provides a cytokine-based culture method for ex vivo expansion of NK cells from postembryonic hematopoietic stem cells into a fully closed, large-scale, cell culture bioprocess. We optimized enrichment of CD34 + cells followed by efficient expansion in gas-permeable cell culture bags. Thereafter, expanded CD34 + cells could be reproducibly amplified and differentiated into CD56 +CD3- NK cell products with a mean expansion of more than 2,000 fold and a purity of >90%. Also provided are collections of cultured cells having specific properties.

    Abstract translation: 本发明提供了一种基于细胞因子的培养方法，用于将NK细胞从胚胎造血干细胞离体扩增成完全封闭的大规模细胞培养生物过程。 我们优化了CD34 +细胞的富集，然后在透气细胞培养袋中有效扩增。 此后，扩增的CD34 +细胞可以可重复扩增，并分化成CD56 + CD3-NK细胞产物，其平均扩增量大于2,000倍，纯度> 90％。 还提供了具有特定性质的培养细胞的集合。

    EX VIVO NK CELL DIFFERENTIATION FROM CD34+ HEMATOPOIETIC CELLS
    2.
    发明申请
    EX VIVO NK CELL DIFFERENTIATION FROM CD34+ HEMATOPOIETIC CELLS 审中-公开
    从CD34 + HEMATOPOIETIC细胞的EX VIVO NK细胞分化

    公开(公告)号：WO2013119118A1

    公开(公告)日：2013-08-15

    申请号：PCT/NL2013/050073

    申请日：2013-02-07

    Applicant: IPD-THERAPEUTICS B.V.

    Inventor： SPANHOLTZ, Jan

    IPC: A61K35/14 ,  C12N5/0783

    CPC classification number: C12N5/0646 ,  A61K35/17 ,  A61K39/39558 ,  C12N2501/125 ,  C12N2501/145 ,  C12N2501/21 ,  C12N2501/22 ,  C12N2501/2302 ,  C12N2501/2306 ,  C12N2501/2307 ,  C12N2501/2312 ,  C12N2501/2315 ,  C12N2501/26 ,  C12N2501/91

    Abstract: The present invention relates to the ex vivo differentiation of NK cells from CD34 + hematopoietic stem cells. Such NK cells and their progenitor cells can be used in therapies of a broad range of malignancies. In the present invention it is shown that IL-12 modulates ex vivo NK cell differentiation. Specific, we achieved significantly higher expression of KIR, CD16 and CD62L in the presence of IL-12 in the cell culture system. The induction of receptor expression by IL-12 occurred predominantly on an augmented population of CD33+NKG2A+ NK cells early during NK cell differentiation. These cells further show enhanced cytolytic activity against MHC class I positive AML targets. In line with the enhanced CD16 expression, IL-12 modulated ex vivo generated NK cells exhibit an improved antibody-dependent-cytotoxicity, using anti CD20 antibody on various B cell targets. Additional to the enhanced expression of CD62L, we show that this cell population consists of a specific chemokine receptor profile. By showing an increased capacity for adhesion to lymphendothelial cells and a specific chemokine receptor profile, we show that IL-12 provided the ex vivo generated NK cells with specific tissue-homing abilities.

    Abstract translation: 本发明涉及NK细胞从CD34 +造血干细胞的离体分化。 这种NK细胞及其祖细胞可用于广泛恶性肿瘤的治疗。 在本发明中，显示IL-12调节离体NK细胞分化。 具体来说，我们在细胞培养系统中，在IL-12存在下，实现了KIR，CD16和CD62L的显着更高的表达。 IL-12受体表达的诱导主要发生在NK细胞分化早期CD33 + NKG2A + NK细胞增殖群体上。 这些细胞进一步显示对MHC I类阳性AML靶标的细胞溶解活性增强。 根据增强的CD16表达，IL-12调节的离体产生的NK细胞在各种B细胞靶上使用抗CD20抗体表现出改善的抗体依赖性细胞毒性。 除了增强CD62L的表达外，我们还显示这种细胞群由特异性趋化因子受体谱组成。 通过显示增加粘附于淋巴细胞上皮细胞和特定趋化因子受体分布的能力，我们显示IL-12提供具有特定组织定位能力的离体产生的NK细胞。

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