Yin Qingsong, Chen Lin, Li Qianyu, Mi Ruihua, Li Yufu, Wei Xudong, Song Yongping
Henan Cancer Hospital, Henan Institute of Hematology, and Cancer Hospital of Zhengzhou University, Zhengzhou, Henan China.
Cancer Cell Int. 2014 Dec 24;14:85. doi: 10.1186/s12935-014-0085-9. eCollection 2014.
T cell-mediated immunity plays an important role in enhancing antitumor response.This study aimed to investigate the changes in the T-lymphocyte subpopulation and to characterize the differential expression pattern of corresponding regulatory genes in peripheral blood mononuclear cells (PBMCs) from diffuse large B cell lymphoma (DLBCL) patients before and after chemotherapy.
A total of 56 DLBCL patients were recruited for analysis of T-cell subset distribution in the peripheral blood using flow cytometry; serum interferon (IFN)-γ and interleukin (IL)-4 levels using enzyme-linked immunosorbent assays; and early growth response protein 1 (EGR-1), T-bet, GATA-binding protein 3 (GATA-3), and transforming growth factor (TGF)-β mRNA levels using quantitative reverse-transcription polymerase chain reaction. Twenty-six healthy subjects served as controls.
The percentage of CD3(+)CD4(+)T lymphocytes in peripheral blood from DLBCL patients was significantly decreased, whereas the percentages of CD3(+)CD8(+)T and CD4(+)CD25(+)T cells were significantly increased compared to those in controls (p < 0.05). Serum levels of IFN-γ and IL-4 were also significantly lower in DLBCL patients than those in controls (p < 0.05), and the levels of EGR-1, T-bet, and GATA-3 mRNA in PBMCs were lower (2.69 ± 1.48, 9.43 ± 2.14, and 20.83 ± 9.05 fold, respectively) in DLBCL patients than those in controls. Furthermore, there was a positive association between the levels of EGR-1 and T-bet mRNA (p = 0.001). However, the level of TGF-β mRNA was significantly increased in DLBCL patients, which was inversely associated with the T-bet mRNA level (p = 0.008), but positively associated with the percentage of T regulatory cells in PBMCs (p = 0.011). After three cycles of chemotherapy, the distribution of T-lymphocyte subsets in DLBCL patients were changed, and the levels of EGR-1, T-bet, and GATA-3 mRNA were significantly increased (p < 0.05) compared to those before chemotherapy.
These results demonstrate the changes in T-lymphocyte subpopulations and the altered expression 34 pattern of the corresponding regulatory genes in PBMCs from DLBCL patients after chemotherapy, which are associated with the response of patients to treatment. The preferential expression of the T-bet gene after chemotherapy was closely correlated with the increased expression of the EGR-1 gene and decreased expression of the TGF-β gene.
T细胞介导的免疫在增强抗肿瘤反应中发挥重要作用。本研究旨在调查弥漫性大B细胞淋巴瘤（DLBCL）患者化疗前后外周血单个核细胞（PBMC）中T淋巴细胞亚群的变化，并表征相应调控基因的差异表达模式。
共招募56例DLBCL患者，采用流式细胞术分析外周血中T细胞亚群分布；采用酶联免疫吸附测定法检测血清干扰素（IFN）-γ和白细胞介素（IL）-4水平；采用定量逆转录聚合酶链反应检测早期生长反应蛋白1（EGR-1）、T-bet、GATA结合蛋白3（GATA-3）和转化生长因子（TGF）-β mRNA水平。26名健康受试者作为对照。
与对照组相比，DLBCL患者外周血中CD3(+)CD4(+)T淋巴细胞百分比显著降低，而CD3(+)CD8(+)T和CD4(+)CD25(+)T细胞百分比显著升高（p < 0.05）。DLBCL患者血清IFN-γ和IL-4水平也显著低于对照组（p < 0.05），PBMC中EGR-1、T-bet和GATA-3 mRNA水平低于对照组（分别低2.69±1.48、9.43±2.14和20.83±9.05倍）。此外，EGR-1和T-bet mRNA水平之间呈正相关（p = 0.001）。然而，DLBCL患者TGF-β mRNA水平显著升高，与T-bet mRNA水平呈负相关（p = 0.008），但与PBMC中调节性T细胞百分比呈正相关（p = 0.011）。三个化疗周期后，DLBCL患者T淋巴细胞亚群分布发生变化，与化疗前相比，EGR-1、T-bet和GATA-3 mRNA水平显著升高（p < 0.05）。
这些结果表明，化疗后DLBCL患者PBMC中T淋巴细胞亚群发生变化，相应调控基因的表达模式改变，这与患者的治疗反应相关。化疗后T-bet基因的优先表达与EGR-1基因表达增加和TGF-β基因表达降低密切相关。
