University of Padova, Dept. of Oncology and Surgical Sciences, Via Gattamelata 64, 35128 Padova, Italy.
J Transl Med. 2010 Nov 22;8:121. doi: 10.1186/1479-5876-8-121.
Recent preclinical adoptive immunotherapy studies in murine models prompt to employ "proper" rather than "as many as possible" antigen-specific T cells to gain better therapeutic results. Ideally, "proper" T cells are poorly differentiated in vitro, but retain the capacity to fully differentiate into effector cells in vivo, where they can undergo long-term survival and strong proliferation. Such requirements can be achieved by modifying culture conditions, namely using less "differentiating" cytokines than IL-2.
To evaluate this issue in human T cell cultures, we exploited a well characterized and clinical-grade protocol finalized at generating EBV-specific CTL for adoptive immunotherapy. In particular, we studied the impact of IL-7, IL-15 and IL-21 compared to IL-2 on different aspects of T cell functionality, namely growth kinetics, differentiation/activation marker expression, cytokine production, and short-term and long-term cytotoxicity.
Results disclosed that the culture modifications we introduced in the standard protocol did not improve activity nor induce substantial changes in differentiation marker expression of EBV-specific CTL.
Our data indicated that the addition of γ-chain cytokines other than IL-2 for the generation of EBV-specific T cell cultures did not produce the improvements expected on the basis of recent published literature. This fact was likely due to the intrinsic differences between murine and human models and highlights the need to design ad hoc protocols rather than simply modify the cytokines added in culture.
最近的小鼠模型临床前过继免疫治疗研究表明，应采用“适当数量”而非“尽可能多”的抗原特异性 T 细胞，以获得更好的治疗效果。理想情况下，“适当数量”的 T 细胞在体外分化程度较低，但在体内仍保留完全分化为效应细胞的能力，在体内它们可以长期存活并进行强烈增殖。通过改变培养条件，即使用比 IL-2 更少的“分化”细胞因子，就可以实现这些要求。
为了在人类 T 细胞培养中评估这个问题，我们利用了一种经过充分表征和临床级别的方案，该方案旨在生成用于过继免疫治疗的 EBV 特异性 CTL。特别是，我们研究了与 IL-2 相比，IL-7、IL-15 和 IL-21 对 T 细胞功能的不同方面（如生长动力学、分化/激活标志物表达、细胞因子产生以及短期和长期细胞毒性）的影响。
结果表明，我们在标准方案中引入的培养物修饰并没有改善 EBV 特异性 CTL 的活性，也没有诱导其分化标志物表达的实质性变化。
我们的数据表明，在生成 EBV 特异性 T 细胞培养物时，添加除 IL-2 以外的 γ 链细胞因子并没有产生基于最近发表的文献所预期的改进。这一事实可能是由于鼠类和人类模型之间的内在差异所致，这突出表明需要设计专门的方案，而不是简单地修改培养物中添加的细胞因子。
