CAJ | 학술논문

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。揹景：腫瘤免疫治療目前在臨床治療中已廣汎開展，但是臨床上經過反複輸註後，由于機體內腫瘤免疫抑製因素的增多齣現瞭治療效果下降的情況，嚴重影響瞭長期治療效果，本實驗通過構建二次成瘤模型儘可能的模擬人體的腫瘤髮生及術後複髮轉移的腫瘤微環境，通過採用外源性臍血特異性Dc疫苗，起到改善體內免疫細胞功能，減少體內免疫抑製因素，從而達到改善腫瘤患者體內免疫微環境，髮揮增彊腫瘤免疫治療效果的臨床治療目的。目的：通過成功構建SCID/Beige裸鼠人結腸癌動物模型，研究臍血來源特異性Dc疫苗對人源化免疫重建荷瘤裸鼠成瘤的免疫抑製作用。方法：分彆選取健康誌願者外週血及正常分娩足月產孕婦臍帶血，利用密度梯度離心法貼壁培養不同來源Dc細胞製備負載人結腸腺癌SW-1116抗原的特異性抗原提呈細胞，促其成熟後收穫；選用健康雄性SCID/Beige裸鼠構建動物模型，將人SW-1116細胞接種于裸鼠腋窩皮下，觀察併確定成瘤。空白對照組無任何預先處理，實驗組、對照組及陰性對照組確定成瘤後自鼠尾靜脈註入健康誌願者外週血PBMC行人免疫細胞微環境重建，在行二次荷瘤前24 h實驗組給予臍血Dc疫苗尾靜脈註射，實驗對照組給予健康誌願者Dc疫苗尾靜脈註射，陰性對照組給予健康誌願者無負載抗原的Dc疫苗尾靜脈註射；經上述處理後在荷瘤裸鼠對側腋窩皮下接種人SW-1116細胞，分彆觀察不同荷瘤組裸鼠二次荷瘤後的一般生活狀態、成瘤時間、成瘤大小及瘤重。結果：空白組裸鼠雙側腫瘤生長快，裸鼠短時間內齣現腫瘤破潰，部分齣現活動差及死亡；經免疫預防處理後荷瘤裸鼠一般生活情況良好，腫瘤生長緩慢，腫瘤體積較小，無破潰、壞死及裸鼠死亡；臍血來源Dc細胞能夠負載人結腸癌細胞株SW-1116抗原，併進一步活化成熟髮揮其專職抗原提成能力；臍血Dc實驗組裸鼠瘤重及瘤終體積平均值明顯小于對照組及陰性對照組，比較統計學差異顯著。結論：臍血特異性DC疫苗能夠改善外週血免疫細胞的免疫功能，促進增彊其自身腫瘤免疫殺傷能力，對瘤細胞生長有明顯抑製，延緩其腫瘤進展及程度，有積極的腫瘤免疫治療作用。
배경：종류면역치료목전재림상치료중이엄범개전，단시림상상경과반복수주후，유우궤체내종류면역억제인소적증다출현료치료효과하강적정황，엄중영향료장기치료효과，본실험통과구건이차성류모형진가능적모의인체적종류발생급술후복발전이적종류미배경，통과채용외원성제혈특이성Dc역묘，기도개선체내면역세포공능，감소체내면역억제인소，종이체도개선종류환자체내면역미배경，발휘증강종류면역치료효과적림상치료목적。목적：통과성공구건SCID/Beige라서인결장암동물모형，연구제혈래원특이성Dc역묘대인원화면역중건하류라서성류적면역억제작용。방법：분별선취건강지원자외주혈급정상분면족월산잉부제대혈，이용밀도제도리심법첩벽배양불동래원Dc세포제비부재인결장선암SW-1116항원적특이성항원제정세포，촉기성숙후수획；선용건강웅성SCID/Beige라서구건동물모형，장인SW-1116세포접충우라서액와피하，관찰병학정성류。공백대조조무임하예선처리，실험조、대조조급음성대조조학정성류후자서미정맥주입건강지원자외주혈PBMC행인면역세포미배경중건，재행이차하류전24 h실험조급여제혈Dc역묘미정맥주사，실험대조조급여건강지원자Dc역묘미정맥주사，음성대조조급여건강지원자무부재항원적Dc역묘미정맥주사；경상술처리후재하류라서대측액와피하접충인SW-1116세포，분별관찰불동하류조라서이차하류후적일반생활상태、성류시간、성류대소급류중。결과：공백조라서쌍측종류생장쾌，라서단시간내출현종류파궤，부분출현활동차급사망；경면역예방처리후하류라서일반생활정황량호，종류생장완만，종류체적교소，무파궤、배사급라서사망；제혈래원Dc세포능구부재인결장암세포주SW-1116항원，병진일보활화성숙발휘기전직항원제성능력；제혈Dc실험조라서류중급류종체적평균치명현소우대조조급음성대조조，비교통계학차이현저。결론：제혈특이성DC역묘능구개선외주혈면역세포적면역공능，촉진증강기자신종류면역살상능력，대류세포생장유명현억제，연완기종류진전급정도，유적겁적종류면역치료작용。
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 anti-tumor 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. hTe 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 Speciifc umbilical blood Dc vaccine, Speciifc 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.