中文摘要：

目的建立食蟹猴单个核淋巴细胞体内荧光示踪方法。探索该方法的细胞采集途径、细胞输注部位和流式检测技术参数。方法从外周血和腋窝淋巴结分别提取分离出单个核淋巴细胞,经CFSE染色后分别将荧光细胞输注食蟹猴外周血和手掌大鱼际下;经过1、2、3 h后从外周血和淋巴结再次提取其中的单个核淋巴细胞,用流式细胞仪检测此样本中荧光细胞数量比例,并优选染色浓度、时长和流式检测技术参数。结果 CFSE最佳染色浓度为1μmol/L,最佳染色时长为15 min;细胞悬液输注1、2、3 h后血管通道组中荧光细胞比例平均值依次为2.37%、0.48%、0.01%,而淋巴管通道中荧光细胞比例显著高于血管通道,平均值依次为12.77%、3.31%、0.07%。结论使用淋巴管道系统进行淋巴细胞的采集示踪,与传统的从血液系统示踪方法比较,可以显著提高可监测靶细胞的数量,可以直观、量化、动态地了解淋巴细胞在体内的变化状态,此方法可重复性高,是一种稳定的食蟹猴体内免疫标记示踪方法。

英文摘要：

Objective To establish an in vivo fluorescent method to trace mononuclear lymphocytes in cynomolgus monkey. Methods Mononuclear lymphocytes were extracted from peripheral blood collected from the femoral vein or sub- clavian vein of cynomolgus monkeys ; meanwhile, the lymphocytes were extracted from the axillary lymph node. The isola- ted cells were labeled with CFSE and then re - infused to the same monkey by intravenously injection to the small saphe- nous vein or subcutaneously to lymphatic vessels in the greater thenar area. Then the monocytes were extracted separately as the same way 1 h, 2 h and 3 h after re - infusion. The proportion of fluorescence - labeled ceils was measured with flow cytometry. Results The best concentration of CFSE was 1 p, mol/L, with staining time of 15 min. The proportions of fluo- rescence - labeled cells in the intravenous injection group were 2. 37% , 0. 48% and 0. 01% , 1 h, 2 h and 3 h after injec- tion, respectively; while the proportions in the subcutaneous lymphatic vessel injection group were 12.77% , 3.31% and 0.07%, respectively. Conclusion Our data show that comparing with the traditional blood system way, the use of the lymphatic system as lymphocytes acquisition and tracing can significantly increase monitored target cells. This experiment successfully labeles mononuclear lymphocytes in cynomolgus monkey and detects the labeled cells after re - infusion at a satisfying proportion, which proves it to be a stable immunization tracing method for primates.