Department of Pharmacology, Faculty of Medicine Siriraj Hospital Mahidol University, Bangkok, Thailand.
Asian Pac J Allergy Immunol. 2017 Jun;35(2):67-74. doi: 10.12932/AP0774.
Cytokine induced killer (CIK) cells are ex-vivo expanded T cells endowed with both T and Natural Killer cell properties. The standard protocol for generation of CIK cells is to culture peripheral blood mononuclear cells (PBMC) in the presence of interferon- gamma (IFN-γ), monoclonal antibody (mAb) against CD3 and interleukin-2 (IL-2). However, this protocol lacks costimulatory signal (CD28), crucial for T cell activation. Herein, the proliferation and functional properties of murine thymocytes derived CIK cells generated with or without costimulatory activation provided by anti-CD28 mAb were examined.
The proportion of CIK (Thy1.2+NK1.1+ and CD8+NK1.1+) cells in culture and the expression of cytotoxic granules (granzyme B and perforin) and proinflammatory cytokines (IFN-γ and tumor necrosis factor-alpha (TNF-α)) were determined by flow cytometry. Additionally, CIK cell cytotoxicity against YAC-1 murine lymphoma cells was measured by a propidium iodide-based assay.
The addition of anti-CD28 to standard CIK culture conditions increased the number of Thy1.2+ NK1.1+ and CD8+ NK1.1+ (the major effector population) cells by almost 40% and 32%, respectively. Furthermore, the cytotoxic potential of CIK cells cultured with the addition of anti-CD28 mAb was also enhanced, with a corresponding increase in CIK cells expressing granzyme B, perforin, IFN-γ and TNF-α.
The addition of anti-CD28 mAb generated more effective murine T cell-derived CIK cells.
细胞因子诱导的杀伤（CIK）细胞是体外扩增的 T 细胞，具有 T 细胞和自然杀伤细胞的特性。CIK 细胞的标准生成方案是在干扰素-γ（IFN-γ）、抗 CD3 单克隆抗体（mAb）和白细胞介素-2（IL-2）存在的情况下培养外周血单个核细胞（PBMC）。然而，该方案缺乏 T 细胞激活所必需的共刺激信号（CD28）。在此，我们研究了在有或没有抗 CD28 mAb 提供的共刺激激活的情况下，生成的鼠源胸腺细胞衍生的 CIK 细胞的增殖和功能特性。
通过流式细胞术测定培养物中 CIK（Thy1.2+NK1.1+和 CD8+NK1.1+）细胞的比例以及细胞毒性颗粒（颗粒酶 B 和穿孔素）和促炎细胞因子（IFN-γ和肿瘤坏死因子-α（TNF-α））的表达。此外，通过碘化丙啶基测定法测量 CIK 细胞对 YAC-1 鼠淋巴瘤细胞的细胞毒性。
在标准 CIK 培养条件中添加抗 CD28 可使 Thy1.2+NK1.1+和 CD8+NK1.1+（主要效应细胞群）细胞的数量分别增加近 40%和 32%。此外，添加抗 CD28 mAb 培养的 CIK 细胞的细胞毒性也得到增强，相应地增加了表达颗粒酶 B、穿孔素、IFN-γ和 TNF-α的 CIK 细胞。
添加抗 CD28 mAb 可生成更有效的鼠源 T 细胞衍生的 CIK 细胞。
