酪氨酸激酶抑制剂对T淋巴细胞生物学特性影响的研究  

Study on the effect of protein tyrosine kinase inhibitors on the biologic characteristics of T lymphocytes

作　　者：许娜[1] 杜庆锋[1] 杨军[1] 刘志[1] 王霜[1] 刘晓力[1] 

机构地区：[1]南方医科大学南方医院血液科,广州510515

出　　处：《国际免疫学杂志》2010年第2期152-156,共5页International Journal of Immunology

基　　金：广州市科技攻关计划重点引导项目（2006Z3-E0401）

摘　　要：目的探讨酪氨酸激酶抑制剂（TKI）在体外对T淋巴细胞生物学特性的影响。方法应用尼龙毛柱法分选人外周血T淋巴细胞，加入植物血凝素和抗人CD3单克隆抗体体外培养；运用MTT法检测不同浓度的伊马替尼（imatinjb）、尼罗替尼（nilotib）对T细胞增殖的影响；酶联免疫吸附试验（ELISA）法检测其分泌的细胞因子浓度；通过流式细胞仪检测T细胞表面免疫活性标志；实时荧光定量PCR法检测各组T细胞信号分子P56^lck。mRNA的表达。结果伊马替尼、尼罗替尼对T淋巴细胞增殖活性有抑制作用（P=0），且呈浓度和时间依赖性；各处理组细胞因子IFN-γ、TNF-α、IL-4、IL-2浓度较对照组低（P=0．03），IL—10浓度在处理组与对照组无差异（P=0．53），且表面活性分子CD25和CD69明显减少（P=0．03，P=0．01），尼戮替尼处理组CD25、CD69表达更低（P=0．05，P=0）；各处理组T细胞信号分子P56^lck。表达水平降低（P=0．03），而尼罗替尼处理组P56^lck。表达更低（P=0．04）。结论有效治疗浓度的伊马替尼、尼罗替尼在体外可能通过抑制T淋巴细胞受体P56^lck的表达进而抑制T细胞的增殖和活性。Objective To study the effects of protein tyrosine kinase Inhibitors on biologic characteristics of T Lymphocytes. Methods T lymphocytes were enriched by nylon wool column from freshly prepared peripheral blood mononuclear cells （PBMC） and cultured in the presence of Anti-CD3 antibody and phytohemagglutinin . Proliferation of T lymphocytes was assayed by MTT method. Cytokine production was measured with a commercial enzyme-linked immnnosorbent assay （ELISA） kit . Effects of imatinib and nilotinib on the function of T lymphocytes were analyzed by flow cytometry . The DOI ： 10. 3760/cma. j. iss. n. 1673-4394. 2010.01. 002 expression of T cell receptor signaling molecule P56^lck was detected by real time-PCR. Results Imatinib and nilotinib obviously supressed the prolifercaion of T lymphocytes in a dose-dependent manner （ P = 0.05 ）. T lymphocytes obtained from cuhure in the presence of imatinib and nilotinib showed reduced expression levels of activation markers CD25 and CD69（P = 0.05, P = 0.01 ） , especially in the presence of nilotinib （ P = 0. 05, P = 0）. Dose-dependent inhibitions of the proliferative response of T lymphocytes observed and associated with reduction such as in IFN-γ,TNF-α,IL-4, and IL-2 productions （P = 0.03 ）, but no significant change in IL-10 aetion（P =0.53 ）. Furthermore , imatinib and nilotinib inhibited P56^lck （P 〈 0. 03 ）, and the level of P56^lck was lower in culture with nilotinib than in imatinib groups （P = 0.04 ）. Conclusion Therapeutic concentrations of imatinib and inlotinib inhibit the activation and proliferation of T lymphocytes, probablely through inhibition of T cell receptor signaling molecule P56^lck.

关 键 词：酪氨酸激酶 伊马替尼 尼罗替尼 T淋巴细胞 P56^lck 

分 类 号：R[医药卫生]