摘要
探讨转染人巨噬细胞炎性蛋白1β(human macrophage inflammatory protein-1 beta,hMIP-1β)基因的小鼠结肠腺癌CT26细胞的生物学特性、致瘤性及对免疫细胞的体内外趋化作用。利用重组腺病毒载体携带hMIP-1β基因(AdhMIP-1β)转染CT26细胞。X-gal染色法检测基因转染效率;ELISA法检测基因转染CT26细胞培养上清中hMIP-1β的含量;Boyden趋化小室法检测培养上清对CD4+T细胞、CD8+T细胞、NK细胞及未成熟树突状细胞(immature dendritic cell,imDC)的趋化作用。小鼠皮下接种转染hMIP-1β基因的CT26细胞,观察其体内致瘤性和对免疫细胞的趋化作用。结果显示,腺病毒载体可携带hMIP-1β基因转染CT26细胞并高效表达hMIP-1β(P<0.01),培养上清含hMIP-1β且对免疫细胞有明显趋化作用。转染hMIP-1β基因的CT26细胞皮下接种后成瘤率降低,肿瘤生长速度减慢,肿瘤内可见明显坏死灶,坏死灶内和周围可见较多淋巴细胞浸润。提示hMIP-1β基因转染CT26细胞后,通过分泌hMIP-1β趋化淋巴细胞等免疫细胞到肿瘤局部发挥有效的抗肿瘤作用,从而为肿瘤的免疫治疗提供新的思路和策略。
This study was to investigate the changes of biological characters and tumorigenicity and the chemotactic activities to immune ceils of tumor ceils transfected with human macrophage inflammatory protein-1 beta（hMIP-1β） gene. Murine colorectal adenocarcinoma CT26 cells were transfected with a recombinant adenovirus carrying the hMIP-1β gene （AdhMIP-1β）. Then the gene transfected was tested by X-gal method and hMIP-1β level in supernatants of hMIP-1β transfected CT26 ceils was assayed by ELISA. The chemotactic activity for CD^4＋ T ceils, CD8^＋ T ceils, NK ceils and immature dendritic ceils （imDC） were assayed by a transwell chamber, and change of growth characterization and in vivo tumorigenicity of hMIP-1β gene-transfected CT26 ceils were also investigated. The results showed that hMIP-1β gene could be transfected into CT26 ceils by AdhMIP-1β efficienctly. The level of hMIP-1β in the culture supernatant of hMIP-1β gene-transfected CT26 ceils was enhanced and the supernatant displayed marked chemotactic activities to CD4＋ T ceils, CD8＋ T ceils, NK ceils and imDC compared with LacZ gene-transfected CT26 ceils and control. When the hMIP-1β gene-transfected CT26 ceils were subcutaneously inoculated to BALB/c mice, tumorigencity was delayed and suppressed, and overt necrosis and lymphocytes infiltration were observed in tumor tissue but not in those inoculated with LacZ gene-transfected CT26 cells or parental CT26 cells. These findings implicate that hMIP-1β gene could be transfected into CT26 cells and be expressed by AdhMIP-1β with high efficiency, and hMIP-1β gene-transfected CT26 cells exhibited decreased tumorigenicity and could induce antitumor immunity. The data provided a basis for further investigation on tumor vaccine transfected with hMIP-1β gene.
出处
《现代免疫学》
CAS
CSCD
北大核心
2007年第1期27-31,共5页
Current Immunology
基金
广西科技厅青年基金资助项目(桂科青0447049)
