特异性脐血DC疫苗对Beige裸鼠人结肠癌动物模型荷瘤免疫治疗作用的实验研究
作者: |
1,2付泽娴,
1,2刘飞,
1,2周晔,
1,2蔡建辉
1 河北医科大学,石家庄050017 2 河北工程大学附属医院,邯郸 056002 |
通讯: | |
DOI: | 10.3978/j.issn.2095-6959.2015.06.S132 |
摘要
背景:肿瘤免疫治疗目前在临床治疗中已广泛开展,但是临床上经过反复输注后,由于机体内肿瘤免疫抑制因素的增多出现了治疗效果下降的情况,严重影响了长期治疗效果,本实验通过构建二次成瘤模型尽可能的模拟人体的肿瘤发生及术后复发转移的肿瘤微环境,通过采用外源性脐血特异性Dc疫苗,起到改善体内免疫细胞功能,减少体内免疫抑制因素,从而达到改善肿瘤患者体内免疫微环境,发挥增强肿瘤免疫治疗效果的临床治疗目的。目的:通过成功构建SCID/Beige裸鼠人结肠癌动物模型,研究脐血来源特异性Dc疫苗对人源化免疫重建荷瘤裸鼠成瘤的免疫抑制作用。方法:分别选取健康志愿者外周血及正常分娩足月产孕妇脐带血,利用密度梯度离心法贴壁培养不同来源Dc细胞制备负载人结肠腺癌SW-1116抗原的特异性抗原提呈细胞,促其成熟后收获;选用健康雄性SCID/Beige裸鼠构建动物模型,将人SW-1116细胞接种于裸鼠腋窝皮下,观察并确定成瘤。空白对照组无任何预先处理,实验组、对照组及阴性对照组确定成瘤后自鼠尾静脉注入健康志愿者外周血PBMC行人免疫细胞微环境重建,在行二次荷瘤前24 h实验组给予脐血Dc疫苗尾静脉注射,实验对照组给予健康志愿者Dc疫苗尾静脉注射,阴性对照组给予健康志愿者无负载抗原的Dc疫苗尾静脉注射;经上述处理后在荷瘤裸鼠对侧腋窝皮下接种人SW-1116细胞,分别观察不同荷瘤组裸鼠二次荷瘤后的一般生活状态、成瘤时间、成瘤大小及瘤重。结果:空白组裸鼠双侧肿瘤生长快,裸鼠短时间内出现肿瘤破溃,部分出现活动差及死亡;经免疫预防处理后荷瘤裸鼠一般生活情况良好,肿瘤生长缓慢,肿瘤体积较小,无破溃、坏死及裸鼠死亡;脐血来源Dc细胞能够负载人结肠癌细胞株SW-1116抗原,并进一步活化成熟发挥其专职抗原提成能力;脐血Dc实验组裸鼠瘤重及瘤终体积平均值明显小于对照组及阴性对照组,比较统计学差异显著。结论:脐血特异性DC疫苗能够改善外周血免疫细胞的免疫功能,促进增强其自身肿瘤免疫杀伤能力,对瘤细胞生长有明显抑制,延缓其肿瘤进展及程度,有积极的肿瘤免疫治疗作用。
关键词:
特异性脐血 DC 疫苗
免疫抑制
Beige裸鼠
结肠癌
The study on specific umbilical blood Dc vaccine for Beige nude mice loaded human colorectal carcinoma to induce anti-tumor immunity
DOI: 10.3978/j.issn.2095-6959.2015.06.S132
Abstract
Background: With the using of tumor immunotherapy in the clinical treatment, a lot of problems were founded such as the factor of the tumor microenvironment, the suppressed antitumor immunocells and the poor long term treatment effect. We built the second load tumor model of SCID/Beige nude mice to recur the tumor microenvironment in beginning of tumor development and recurrence. Specific umbilical blood Dc vaccine was used to stimulate the function of the immunocells and reduce the inhibition of tumor immune effector cells in tumor microenvironment. Objective: The SCID/Beige nude mice which second loaded human colon tumor cells, namely SW-1116 cells, were used to build the models. To observe the immunosuppression of specific umbilical blood Dc vaccine to tumor development of SCID/ Beige nude mice which handled with human PBMCs. Methods: Samples of peripheral blood in volunteers and umbilical blood in the childbirth pregnant women were collected to enrich Dcs by density gradient centrifugation. When the specific Dc vaccines matured they were gained, which hatched with human colon tumor SW-1116 cells. The male SCID/Beige nude mice were used to build model, which subcutaneous injected with human SW-1116 cells in axillary of nude mice. The nude mice were observed to make sure that tumor were formed. The mice of the blank group were handled with nothing. That of the Specific umbilical blood Dc vaccine group (ubDc), the Specific peripheral blood Dc vaccine group (pbDc), and The naked peripheral blood Dc vaccine group (npbDc) were injected with human peripheral blood mononuclear cells to recur the humanized immune reconstruction. Twenty four hours before injecting tumor cells, the mice of different groups were received the treatment with Specific umbilical blood Dc vaccine, Specific peripheral blood Dc vaccine and the naked peripheral blood Dc vaccine respectively. The general life, tumor growing time, tumor size and weight of the mice were observed after they were given a suspension of human SW-1116 cells subcutaneously in contralateral axillary. Results: The nude mice of blank group were found the bilateral tumor fast growth and burst, even part of them was poor and death. On the contrary, the nude mice protected by the immune prevention treatment were found the tumor slow growth, small tumor size and no burst or necrosis and death. Human umbilical cord blood Dc can load human SW-1116 antigen and matured as the antigen presenting cells. The tumor weights and volumes of the ubDc group were smaller than other groups. It is compared statistically significant. Conclusion: The specific umbilical cord blood Dc vaccine can improve the immune function of peripheral immune cells and promote to improve their immune ability to kill tumor cells. They also have obvious inhibition on tumor cell growth and delay the tumor development. The specific umbilical cord blood Dc vaccine plays the key role in tumor immunotherapy.