新城疫病毒通过上调小鼠NK细胞TRAIL表达杀伤Novikoff小鼠肝癌细胞

Newcastle disease virus enhances tumoricidal activity of mouse NK cells against mouse Novikoff hepatoma cells via up-regulating expression of TRAIL on the NK cells

目的观察经腹腔注射的新城疫病毒(NDV)对小鼠脾脏NK细胞表达肿瘤坏死因子相关凋亡诱导配体(TRAIL)及杀伤Novikoff小鼠肝癌细胞的影响,并探讨其与γ干扰素(IFN-γ)的关系。方法给予BALB/c小鼠和IFN-γR-/-B6.129S7小鼠腹腔注射NDV,12 h后,用ELISA检测小鼠外周血IFN-γ浓度;分离小鼠脾脏NK细胞,用反转录PCR法检测NK细胞中TRAIL mRNA转录水平,Western blot法检测脾脏NK细胞中TRAIL蛋白水平,用乳酸脱氢酶(LDH)释放法检测NK细胞对Novikoff肝癌细胞的杀伤作用。结果腹腔注射NDV可以提高BALB/c小鼠外周血IFN-γ浓度,上调小鼠脾脏NK细胞TRAIL mRNA和蛋白表达水平,增强小鼠脾脏NK细胞体外条件下对Novikoff肝癌细胞的杀伤活性。TRAIL中和抗体能抑制NK细胞对Novikoff肝癌细胞的杀伤作用。IFN-γR-/-B6.129S7小鼠接受NDV注射后脾脏NK细胞的TRAIL表达无显著增加,NDV激活的NK细胞对Novikoff肝癌细胞的杀伤活性显著低于IFN-γ受体正常的BALB/c小鼠。结论腹腔注射NDV可增强脾脏NK细胞的体外抗肿瘤活性,其机制之一是通过IFN-γ受体途径上调NK细胞表面TRAIL蛋白表达来发挥作用。

Objective To observe the effect of intraperitoneal injection of Newcastle disease virus （NDV） on tumor necrosis factor-related apoptosis-inducing ligand （TRAIL） expression in mouse spleen NK cells and NK cells-mediated tumoricidal activity against mouse Novikoff hepatoma cell line, and explore the role of interferon （ IFN）-γ in NDV-induced TRAIL expression and tumodcidal activity. Methods NDV was injected intraperitoneaUy to BALB/c mice and IFN-y receptor- deficient （IFN-γR-/-） [36. 129S7 mice. Twelve hours after injection, the concentration of IFN-γ in peripheral blood from BALB/c mice was determined by ELISA. Mouse spleen NK cells were separated. The mRNA and protein expression of TRAIL in NK cells were detected through reverse transcription PCR （RT-PCR） and Western blotting. Lactate dehydrogenase （LDH） release assay was used to determine the cytotoxic activity of NK cells against mouse hepatoma cells. Results NDV injection increased the IFN-γ concentration in peripheral blood of BALB/c mice, induced up-regulation of TRAIL at the mRNA and protein levels in mouse spleen NK cells, and enhanced the killing ability of mouse spleen NK cells towards Novikoff hepatoma cells. Blocking TRAIL by neutralizing antibody suppressed the cytotoxic activity of NK cells against Novikoff hepatoma cells. Furthermore, NDV injection in IFN-γ R-/- B6. ]29S7 mice did not make significant difference from control group in TRAIL expression in spleen NK cells, and the tumoricidal activity of IFN-7,R-γ- I%. 129S mouse spleen NK cells against Novikoff hepatoma cells was significantly lower than that of BALB/c mouse NK cells. Conclusion Intraperitoneal injection with NDV could enhance tumoricidal activity of mouse spleen NK cells in v/tro, and one of the mechanisms might be that NDV injection up-regulates TRAIL expression in NK cells through the IFN-）, receptor pathway.