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METHOD FOR PREPARING GENETICALLY-MODIFIED T CELLS WHICH EXPRESS CHIMERIC ANTIGEN RECEPTOR

机译:表达嵌合抗原受体的基因修饰的T细胞的制备方法

摘要

In order to improve the efficiency of gene introduction in CAR therapy employing a transposon method, provided is a method for preparing genetically-modified T cells expressing chimeric antigen receptor, comprising: (1) a step of preparing non-proliferative cells which are obtained by stimulating a group of cells comprising T cells using an anti-CD3 antibody and an anti-CD28 antibody followed by a treatment for causing the cells to lose their proliferation capability; (2) a step of obtaining genetically-modified T cells into which a target antigen-specific chimeric antigen receptor gene has been introduced using a transposon method; (3) a step of mixing the non-proliferative cells prepared by step (1) with the genetically-modified T cells obtained by step (2), and co-culturing the mixed cells while stimulating the mixed cells using an anti-CD3 antibody and anti-CD28 antibody; and (4) a step of collecting the cells after culture. Also, provided is a method for preparing genetically-modified T cells expressing chimeric antigen receptor, comprising: (i) a step of preparing non-proliferative cells holding a viral peptide antigen, which are obtained by stimulating a group of cells comprising T cells using an anti-CD3 antibody and an anti-CD28 antibody followed by culturing in the presence of the viral peptide antigen and a treatment for causing the cells to lose their proliferation capability; (ii) a step of obtaining genetically-modified T cells into which a target antigen-specific chimeric antigen receptor gene has been introduced using a transposon method; (iii) a step of mixing the non-proliferative cells prepared by step (i) with the genetically-modified T cells obtained by step (ii), and co-culturing the mixed cells; and (iv) a step of collecting the cells after culture.
机译:为了提高在采用转座子方法的CAR疗法中基因导入的效率,提供了一种制备表达嵌合抗原受体的基因修饰的T细胞的方法,其包括:(1)制备通过下述方法获得的非增殖细胞的步骤。使用抗CD3抗体和抗CD28抗体刺激由T细胞组成的一组细胞,然后进行使细胞丧失增殖能力的处理。 (2)利用转座子法获得导入了靶抗原特异性嵌合抗原受体基因的基因修饰T细胞的步骤。 (3)将通过步骤(1)制备的非增殖性细胞与通过步骤(2)获得的基因修饰的T细胞混合的步骤,在使用抗CD3抗体刺激混合细胞的同时进行混合培养和抗CD28抗体; (4)培养后收集细胞的步骤。另外,提供了一种制备表达嵌合抗原受体的基因修饰的T细胞的方法,该方法包括:(i)制备含有病毒肽抗原的非增殖性细胞的步骤,所述非增殖性细胞是通过刺激一组包含T细胞的细胞而获得的。抗CD3抗体和抗CD28抗体,然后在病毒肽抗原的存在下培养,并进行使细胞丧失增殖能力的处理。 (ii)使用转座子法获得已导入了靶抗原特异性嵌合抗原受体基因的基因修饰T细胞的步骤; (iii)将步骤(i)制备的非增殖细胞与步骤(ii)获得的基因修饰的T细胞混合,并共同培养的步骤; (iv)培养后收集细胞的步骤。

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