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目的研究脐血源性T淋巴细胞在小鼠体内的存活、浸润情况,以及对红白血病(EL)小鼠的治疗作用,为探讨脐血源性T淋巴细胞的生物学特性及其治疗白血病提供实验和理论依据。方法48只BALB/c小鼠随机分为对照组、实验1组和实验2组。分离制备脐血源性T淋巴细胞,对其标记后经尾静脉或腹腔移植入红白血病BALB/c小鼠体内,采用免疫荧光、免疫组织化学、透射电镜等方法观察脐血源性T淋巴细胞在小鼠体内的存活时间、浸润状况及其对白血病小鼠脾脏结构的影响。结果脐血单个核细胞(CBMNCs)经植物血凝集素(PHA)诱导后,绝大多数细胞为CD3+T淋巴细胞,阳性率可达(83.42±1.26)%;移植后第3、7天均可在小鼠外周血和肝、脾组织中检测到人CD3+细胞和Brdu+细胞;两实验组(EG)小鼠肝、脾组织内均见CD25+细胞散在浸润;两实验组EL小鼠平均存活时间分别为(27.25±7.06)d和(24.74±2.93)d,与对照组比较,均有统计学意义(P<0.05);Hoechst33258染色法检测显示,在荧光显微镜下见对照组细胞核荧光均匀,凋亡细胞较少;实验组部分白血病细胞呈现凋亡的形态学改变:染色质高度凝聚,致密浓染,或裂解成碎块状,边缘光滑清晰;透射电镜下见对照组小鼠脾内白血病细胞广泛浸润,实验组小鼠部分白血病细胞呈现典型凋亡形态学改变:核染色质浓缩、边集;核碎裂,可见凋亡小体,粗面内质网(RER)扩张,线粒体(Mi)呈现空泡样变及髓样变。结论脐血源性T淋巴细胞能在白血病小鼠体内存活并趋化至白血病细胞浸润部位,发挥其相应的功能,对白血病有一定的治疗作用。
Objective To study the survival and infiltration of umbilical cord blood-derived T lymphocytes in mice and the therapeutic effect on erythroleukemia (EL) mice. To explore the biological characteristics of cord blood-derived T lymphocytes and their treatment for leukemia Experimental and theoretical basis. Methods 48 BALB / c mice were randomly divided into control group, experimental group 1 and experimental group 2. Umbilical cord blood-derived T lymphocytes were isolated and labeled, and then labeled into tail vein or abdominal cavity into erythroleukemia BALB / c mice. Immunofluorescence, immunohistochemistry and transmission electron microscopy were used to observe the expression of umbilical cord blood-derived T lymphocytes Survival time in mice, infiltration and its effect on the spleen structure of leukemia mice. Results After CBMNCs were induced by phytohemagglutinin (PHA), most of the cells were CD3 + T lymphocytes, the positive rate was (83.42 ± 1.26)%. On the 3rd and 7th days after transplantation Both CD3 + cells and Brdu + cells could be detected in the peripheral blood of mice and liver and spleen tissues. CD25 + cells scattered in the liver and spleen of two experimental groups (EG) (27.25 ± 7.06) d and (24.74 ± 2.93) d respectively, which were statistically significant compared with the control group (P <0.05). Hoechst33258 staining showed that the fluorescence intensity of the control group was uniform, Apoptotic cells were less; some leukemic cells in the experimental group showed morphological changes of apoptosis: the chromatin was highly condensed, densely stained, or cracked into pieces, the edge was smooth and clear; and the control group mice splenic leukemia The cells were extensively infiltrated. Some leukemia cells in the experimental group showed morphological changes of typical apoptosis: nuclear chromatin condensation, edge set; nuclear fragmentation, apoptotic bodies, RER expansion, mitochondrial (Mi ) Showed vacuolar and myeloid degeneration. CONCLUSION: Umbilical cord blood-derived T lymphocytes can survive in leukemia mice and chemotaxis to the leukemia cells infiltration site, play their corresponding functions, and have a certain therapeutic effect on leukemia.