Development of a clinical model for ex vivo expansion of multiple populations of effector cells for adoptive cellular therapy
Development of a clinical model for ex vivo expansion of multiple populations of effector cells for adoptive cellular therapy
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DOI：
10.1080/14653240701762398
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发表时间：
2008-01-01
期刊：
影响因子：
4.5
通讯作者：
Sentman, C.
中科院分区：
文献类型：
--
作者：
Meehan, K. R.;Wu, J.;Sentman, C.
背景我们先前已经证明了一种将自体单核细胞扩增为效应杀伤细胞群体的实验室模型。本实验的目的是开发一种良好的生产规范（GMP）方法，用于扩增临床级活化的效应细胞，所述效应细胞通过各种机制介导肿瘤细胞杀伤，所述效应细胞可以在高剂量化疗和自体干细胞移植后输注到患者体内。白细胞介素-2（1000 IU/mL）和OKT-3（500 ng/mL）在37 ℃和5%CO2条件下。扩增7天后，分析细胞的细胞浓度、活力、表型和针对人骨髓瘤细胞系的细胞毒性。使用培养袋和烧瓶比较扩增。冷冻保存的扩增细胞也analysed.ResultsThis离体扩增的临床模型产生的细胞毒性淋巴细胞，包括CD 3 + CD 4 + T细胞，CD 3 + CD 8 + T细胞，CD 8 + CD 56 + T细胞和CD 56+自然杀伤细胞的多克隆群体。与烧瓶相比，培养袋提供了2-3倍的效应细胞扩增，污染风险最小。扩增时的最佳细胞浓度为2.5-3.5 × 106外周血MNC/mL。如果扩增的细胞被冷冻保存，然后解冻use.DiscussionThe结果表明，目前正在临床试验中采用的可重复和可靠的GMP程序，保持活力和细胞毒性。这些扩增的细胞及其杀死肿瘤细胞的各种途径可能会绕过肿瘤逃逸机制并改善结果。
BackgroundWe have previously demonstrated a laboratory model for expanding autologous mononuclear cells into populations of effector killer cells. The goal of the current experiments was to develop a good manufacturing practice (GMP) method for expanding clinical-grade activated effector cells that mediate tumor cell killing through various mechanisms that could be infused into patients following high-dose chemotherapy and autologous stem cell transplant.MethodsMobilized mononuclear cells (MNC) from myeloma patients were placed in culture with serum-free AIM V media, interleukin-2 (1000 IU/mL) and OKT-3 (500 ng/mL) at 37C and 5% CO2. After 7 days of expansion, the cells were analyzed for cell concentration, viability, phenotype and cytotoxicity directed against human myeloma cell lines. Expansion was compared using culture bags and flasks. Cryopreserved expanded cells were also analyzed.ResultsThis clinical model of ex vivo expansion yielded polyclonal populations of cytotoxic lymphocytes, including CD3+ CD4+ T cells, CD3+ CD8+ T cells, CD8+ CD56+ T cells and CD56+ natural killer cells. Compared with flasks, culture bags provided a 2-3-fold effector cell expansion with minimal risk of contamination. The optimal cell concentration at the time of expansion was 2.5-3.5 106 peripheral blood MNC/mL. Viability and cytotoxicity were maintained if the expanded cells were cryopreserved and then thawed for use.DiscussionThe results demonstrate a reproducible and reliable GMP procedure that is currently being employed in a clinical trial. These expanded cells, and their various pathways of tumor cell killing, may circumvent tumor escape mechanisms and improve outcomes.