Institute of Hematology, Medical College, Jinan University, Guangzhou, China.
DNA Cell Biol. 2012 Nov;31(11):1628-35. doi: 10.1089/dna.2012.1798. Epub 2012 Oct 11.
T-cell immunodeficiency is a common feature in patients with chronic myeloid leukemia (CML), and deficiency in CD3 levels was detected in T cells from these patients, which may represent a characteristic that is related to a lower T cell activation. In this study, we explored the possibility that forced TCRζ gene expression may upreg-u-late T cell receptor (TCR) signaling activation and reverse interleukin-2 (IL-2) production in T cells from patients with CML. A recombinant eukaryotic vector expressing TCRζ was transfected into T cells by nucleofection. Phosphorylated TCRζ, phosphorylated NF-κB, and the IL-2 level in TCRζ-transfected CD3+T cells that were activated with anti-CD3 and anti-CD28 antibodies were measured by Western blot and enzyme-linked immunosorbent assay (ELISA). Significantly increased TCRζ levels were found in TCRζ-transfected CD3+T cells. After CD3 and CD28 antibody stimulation, a significantly higher phosphorylated TCRζ chain level was demonstrated, and an increased IL-2 production in TCRζ-upregulated T cells was associated with the increased expression of the phosphorylated NF-κB. In conclusion, TCRζ gene transfection could restore TCRζ chain deficiency and enhance IL-2 production in T cells from patients with CML. It is possible that TCRζ chain reconstitution in leukemia-specific, clonally expanded T cells will effectively increase their activation of antileukemia cytotoxicity.
T 细胞免疫缺陷是慢性髓系白血病(CML)患者的常见特征,这些患者的 T 细胞中检测到 CD3 水平缺乏,这可能代表与较低的 T 细胞激活相关的特征。在这项研究中,我们探讨了强制表达 TCRζ 基因是否可能上调 T 细胞受体(TCR)信号激活并逆转 CML 患者 T 细胞中白细胞介素-2(IL-2)的产生。通过核转染将表达 TCRζ 的重组真核载体转染到 T 细胞中。通过 Western blot 和酶联免疫吸附试验(ELISA)测量用抗-CD3 和抗-CD28 抗体激活的转染 TCRζ 的 CD3+T 细胞中磷酸化 TCRζ、磷酸化 NF-κB 和 IL-2 水平。在转染 TCRζ 的 CD3+T 细胞中发现 TCRζ 水平显著增加。在 CD3 和 CD28 抗体刺激后,磷酸化 TCRζ 链水平明显升高,TCRζ 上调的 T 细胞中 IL-2 的产生增加与磷酸化 NF-κB 的表达增加相关。总之,TCRζ 基因转染可以恢复 CML 患者 T 细胞中 TCRζ 链的缺乏并增强 IL-2 的产生。在白血病特异性、克隆扩增的 T 细胞中重建 TCRζ 链可能会有效地增加它们对白血病细胞毒性的激活。