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目的通过体内外实验研究新城疫病毒(NDV)对人急性淋巴细胞白血病的作用。方法体外实验应用倒置显微镜观察NDV感染后白血病细胞形态和密度变化,四甲基偶氮唑蓝(MTT)法测定不同水平NDV作用于白血病细胞不同时间后对细胞增殖的影响。体内实验通过建立人急性白血病裸鼠异种移植模型,瘤内注射NDV,测量瘤体大小,计算抑瘤率,同时对肿瘤组织病理切片进行HE染色检查。并通过对裸鼠一般状态的观察及行肝脾病理切片检查评估NDV在体内的不良反应。结果 1.倒置显微镜下,对照组及尿囊液组细胞形态规则、分布密集;NDV感染组细胞形态不规则,聚集、融合现象明显,分布稀疏。2.MTT检测结果显示不同水平NDV对T淋巴细胞白血病细胞(MOLT-4)的增殖均有明显抑制作用(P<0.01),不同水平间NDV的抑制作用比较,差异有统计学意义(P<0.01);不同时间的抑制作用比较差异亦有统计学意义(P<0.01);增殖抑制作用呈时间-浓度依赖关系。3.NDV原液对MOLT-4裸鼠异种移植瘤的抑制率为86.2%;肿瘤质量与对照组比较差异有统计学意义(P<0.01)。4.肿瘤组织病理切片HE染色可见大片坏死区,细胞结构消失。5.实验过程中裸鼠无死亡,一般状态良好,无不良反应。结论 NDV是一种有效的抗人急性白血病病毒,在体内及体外均具有抑制白血病细胞增殖的作用,并且具有很好的安全性。
Objective To study the effect of Newcastle disease virus (NDV) on human acute lymphoblastic leukemia in vitro and in vivo. Methods The morphology and density of leukemia cells after NDV infection were observed by inverted microscope in vitro. The effects of different levels of NDV on the proliferation of leukemia cells were determined by MTT assay. In vivo, by establishing a xenograft model of human acute leukemia in nude mice, NDV was injected intratumorally and tumor size was measured to calculate tumor inhibition rate. At the same time, HE staining was performed on tumor tissue sections. And through the observation of the general state of nude mice and pathological examination of liver and spleen pathological evaluation of NDV in vivo adverse reactions. Results 1. Under the inverted microscope, the cells in the control group and the allantoic fluid group were regular and densely distributed. The cells in the NDV infection group were irregular in shape, aggregated and fused, with sparse distribution. The results of MTT assay showed that different levels of NDV significantly inhibited the proliferation of T lymphoblastic leukemia cells (MOLT-4) (P <0.01), and the inhibitory effects of different levels of NDV were statistically different (P < 0.01). The inhibitory effect at different time was also statistically significant (P <0.01). The proliferation inhibition was time-dependent. The inhibition rate of NDV stock solution on xenograft tumor of MOLT-4 nude mice was 86.2%. There was significant difference between tumor weight and control group (P <0.01). 4. Histopathological sections of the tumor visible necrosis of large areas, cell structure disappeared. Nude mice during the experiment without death, the general state of good, no adverse reactions. Conclusion NDV is a potent anti-human acute leukemia virus that inhibits the proliferation of leukemia cells both in vitro and in vivo and has good safety.