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IL-18是新近发现的细胞因子,可诱生IFN-2的产生、增强NK活性和Th1型免疫反应具有抗肿瘤作用.本研究的目的是研究转染IL-18基因的DC以肿瘤抗原肽冲击致敏后对转移性肺癌的治疗作用.首先从C57BL/6小鼠骨髓培养DC,按MOI=100进行体外转染AdIL-18及AdLacZ基因,X-gal染色提示转染效率大于95%;转染smIL-18后24h、48h,ELISA测定上清中IL-18因子水平分别为(45±8)ng/ml和(54±7)ng/ml;以RT-PCR方法可检测到含插入信号肽的IL-18mRNA的表达,对照DC上清中则未检出;测定DC-IL-18培养上清的活性发现IL-18基因转染之DC上清能显著刺激ConA活化的小鼠脾脏T细胞产生IFN-γ,且与IL-12有协同作用,对照组DC上清则无此作用.FACS检测msIL-18基因修饰前后DC表型发现,提示msIL-18基因修饰后24h,荧光强度无明显变化,但la,B7-2,
IL-18 is a recently discovered cytokine that induces IFN-2 production, enhances NK activity, and has an anti-tumor effect on Th1 immune responses. The aim of this study was to study DCs transfected with the IL-18 gene as tumor antigen peptides. Treatment of metastatic lung cancer after impact sensitization. DCs were first cultured from C57BL/6 mouse bone marrow and then transfected with AdIL-18 and AdLacZ gene at a MOI of 100. X-gal staining indicated that the transfection efficiency was greater than 95%. At 24h and 48h after transfection with smIL-18, IL-18 levels in the supernatant were determined by ELISA at (45±8) ng/ml and (54±7) ng/ml, respectively; the insertion was detected by RT-PCR. The expression of signal peptide IL-18 mRNA was not detected in the control DC supernatant; the activity of DC-IL-18 culture supernatant was determined to find that the DC supernatant transfected with IL-18 gene can significantly stimulate ConA-activated mouse spleen T cells produce IFN-γ and have a synergistic effect with IL-12. The DC supernatant of the control group does not have this effect. The DC phenotype before and after the modification of msIL-18 gene was detected by FACS, suggesting that the fluorescence intensity was 24 h after the modification of msIL-18 gene. No significant change, but la, B7-2,