新城疫病毒对人急性淋巴细胞白血病的作用

Effects of Newcastle Disease Virus on Human Acute Lymphoblastic Leukemia

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 effects of newcastle disease virus （NDV） on human acute lymphoblastic leukemia in vitro and in vivo.Methods In vitro,density and morphologic change of cells infected by NDV were observed by inverted microscope.Methyl thiazolyl tetrazolium （MTT） assay was utilized to observe the effect of NDV on proliferation of the leukemic cells by different concentration and time.In vivo,the xenograft model of human acute leukemia was established in CD-1 nude mice.The effect of NDV by injecting directly into the tumor tissue was evaluated by the size of tumor,the inhibition ratio and the pathological section with hematoxylin and eosin（HE）stain of tumor.The safety of NDV was assessed by the general condition and the liver and spleen pathological section with HE stain of the nude mice.Results 1.The cells of the control and allantoic fluid group were all regular and coarctate by inverted microscope.The effect of NDV were conspicuous in that cells were irregular,sparsate,aggregate and fused in experimental groups.2.MTT assay showed that the proliferation of human acute lympholastic leukemia cell line（MOLT-4） cells were significantly inhibited by NDV with different concentration（P〈0.01）.The results also demonstrated that the difference within the concentration and the time were both statistically significant（Pa0.01）.The inhibition of NDV was further confirmed in a time-concentration-dependent manner.3.The inhibition ratio of NDV stock solution to the growth of MOLT-4 was 86.2%.The different average weight of MOLT-4 xenograft were significant in statistics between the experimental and control group（P〈0.01）.4.The histological changes of cellular structure disappearance and tissue necrosis were observed by light microscope in experimental group with HE stain.5.During the experiment no nude mouse died,no sign of toxicity was found.Conclusions Newcastle disease virus can inhibited human acute leukemic cell proliferation in vitro and in vivo.The safety of NDV is reliable.