Cappel Claudia, Huenecke Sabine, Suemmerer Anica, Erben Stephanie, Rettinger Eva, Pfirrmann Verena, Heinze Annekathrin, Zimmermann Olga, Klingebiel Thomas, Ullrich Evelyn, Bader Peter, Bremm Melanie
Department for Stem Cell Transplantation and Immunology, Clinic for Children and Adolescents, University Hospital Frankfurt, Goethe University, Frankfurt/Main, Germany.
Pediatr Blood Cancer. 2016 Dec;63(12):2230-2239. doi: 10.1002/pbc.26147. Epub 2016 Jul 19.
Neuroblastoma (NB) is the most common solid extracranial tumor in childhood. Despite advances in therapy, the prognosis is poor and optimized therapies are urgently needed. Therefore, we investigated the antitumor potential of interleukin-15 (IL-15)-activated cytokine-induced killer (CIK) cells against different NB cell lines.
CIK cells were generated from peripheral blood mononuclear cells by the stimulation with interferon-γ (IFN-γ), IL-2, OKT-3 and IL-15 over a period of 10-12 days. The cytotoxic activity against NB cells was analyzed by nonradioactive Europium release assay before and after blocking of different receptor-ligand interactions relevant in CIK cell-mediated cytotoxicity.
The final CIK cell products consisted in median of 83% (range: 75.9-91.9%) CD3 CD56 T cells, 14% (range: 5.2-20.7%) CD3 CD56 NK-like T cells and 2% (range: 0.9-4.8%) CD3 CD56 NK cells. CIK cells expanded significantly upon ex vivo stimulation with median rates of 22.3-fold for T cells, 58.3-fold for NK-like T cells and 2.5-fold for NK cells. Interestingly, CD25 surface expression increased from less than equal to 1% up to median 79.7%. Cytotoxic activity of CIK cells against NB cells was in median 34.7, 25.9 and 34.8% against the cell lines UKF-NB-3, UKF-NB-4 and SK-N-SH, respectively. In comparison with IL-2-stimulated NK cells, CIK cells showed a significantly higher cytotoxicity. Antibody-mediated blocking of the receptors NKG2D, TRAIL, FasL, DNAM-1, NKp30 and lymphocyte function-associated antigen-1 (LFA-1) significantly reduced lytic activity, indicating that diverse cytotoxic mechanisms might be involved in CIK cell-mediated NB killing.
Unlike the mechanism reported in other malignancies, NKG2D-mediated cytotoxicity does not constitute the major killing mechanism of CIK cells against NB.
神经母细胞瘤（NB）是儿童期最常见的实体颅外肿瘤。尽管治疗取得了进展，但预后仍然很差，迫切需要优化治疗方法。因此，我们研究了白细胞介素-15（IL-15）激活的细胞因子诱导杀伤细胞（CIK）对不同NB细胞系的抗肿瘤潜力。
通过用干扰素-γ（IFN-γ）、IL-2、OKT-3和IL-15刺激外周血单个核细胞10 - 12天来生成CIK细胞。在阻断CIK细胞介导的细胞毒性中相关的不同受体 - 配体相互作用之前和之后，通过非放射性铕释放试验分析对NB细胞的细胞毒性活性。
最终的CIK细胞产物中，CD3 CD56 T细胞中位数为83%（范围：75.9 - 91.9%），CD3 CD56 NK样T细胞为14%（范围：5.2 - 20.7%），CD3 CD56 NK细胞为2%（范围：0.9 - 4.8%）。CIK细胞在体外刺激后显著扩增，T细胞的中位数扩增率为22.3倍，NK样T细胞为58.3倍，NK细胞为2.5倍。有趣的是，CD25表面表达从小于等于1%增加到中位数79.7%。CIK细胞对NB细胞系UKF - NB - 3、UKF - NB - 4和SK - N - SH的细胞毒性活性中位数分别为34.7%、25.9%和34.8%。与IL - 2刺激的NK细胞相比，CIK细胞表现出显著更高的细胞毒性。抗体介导的对受体NKG2D、TRAIL、FasL、DNAM - 1、NKp30和淋巴细胞功能相关抗原 - 1（LFA - 1）的阻断显著降低了裂解活性，表明多种细胞毒性机制可能参与CIK细胞介导的NB杀伤。
与其他恶性肿瘤中报道的机制不同，NKG2D介导的细胞毒性不是CIK细胞对NB的主要杀伤机制。
