氦氖激光照射胸腺区促进荷瘤小鼠T细胞的CD59表达与活化信号转导

He-Ne Laser Irradiation to the Thymus Area Enhances CD59 Expression and Activation of Cell Signaling in T Cells of Tumor-Bearing Mice

探讨低功率氦氖激光照射对荷瘤小鼠T细胞CD59表达及其功能的影响。将(BALB/C)小鼠分为4组:Ⅰ组不做处理;Ⅱ组仅致瘤;Ⅲ组致瘤后每日照射胸腺区;Ⅳ组致瘤后隔日照射胸腺区。致瘤后第20天处死小鼠,分离培养胸腺T细胞。采用噻唑蓝(MTT)比色法测T细胞增殖,用免疫组化和免疫荧光测T细胞CD59表达量,然后用酶联免疫吸附测定法(ELISA)测T细胞活化后白介素2(IL-2)水平,用激光共聚焦测T细胞胞浆内钙离子变化,以及用流式细胞术测T细胞活化为CD2+5T细胞的能力。结果表明,照射组肿瘤生长受抑;荷瘤组T细胞的CD59表达量、白介素2水平、增殖能力、钙离子浓度及活化的CD2+5T细胞水平均低于正常组,但照射组明显高于非照射组。由此可见,激光照射促进了荷瘤小鼠T细胞CD59的表达,增强了其活化信号的转导。

The effect of low-power He-Ne laser irradiation on the function and expression of CD59 on T cells in tumor-bearing mice is studied. 40 BALB/C mice were randomly and evenly divided into four groups：tumor-bearing mice were constructed in groupⅡⅢⅣ,while groupⅠwere treated as normal control. Thymus region of mice in group Ⅲ were irradiated every day and mice in group Ⅳ were irradiated every two days. The growth of tumors were noted by tumor growth curves. The mice were executed 20 days later and T cells were segregated from thymus. The expression of CD59 on T cells from thymus was detected by immunohistochemistry and immunofluorescence;lymphocyte proliferation in thymus was detected by lymphocyte transformation test （MTT assay）;IL-2 was detected by enzyme-linked immunosorbent array （ELISA）;Laser scanning confocal microscope imaging of the calcium fluorescent indicator dye Fluo-3/AM was used to determine the kinetic changes of [Ca^2＋] in T cell endochylema. CD25＋ T cells were determined by flow cytometer .The experiments show that tumor growth was inhibited after laser irradiation.The expression of CD59 protein,the proliferation of lymphocytes,level of IL-2,degree of the [Ca^2＋] and the expression of CD25 in group Ⅱ,Ⅲ,Ⅳ were decreased compared with group Ⅰ,but were increased in group Ⅲ,Ⅳcompared with group Ⅱ. The results indicate that the low expression of CD59 was improved after irradiation by low-power He-Ne laser,and irradiation can enhance the function of T cells.