【作者】 李军；

【导师】 张锦堃；

【作者基本信息】 汕头大学 ， 肿瘤病理学， 2002， 硕士

【摘要】 目的 研制小鼠肝癌H₂₂细胞与脾树突状细胞（DC）融合的杂交瘤苗H₂₂-DC，进而观察H₂₂-DC主动诱导体内特异性抗肿瘤免疫反应以及对H₂₂荷瘤小鼠的治疗作用。 方法 以BALB／C小鼠为实验动物，分离脾DC，采用细胞因子诱导、CDllc磁珠标记、MACS分选等技术与传统的聚乙二醇（PEG）法相结合的方法，将DC与H₂₂细胞相融合制备H₂₂-DC杂交瘤苗。并进行体内致瘤性检测。体内抗肿瘤实验分成免疫保护组和免疫治疗组两大组。免疫保护组随机分成P、H、D和HD4小组，分别用PBS、灭活的H₂₂、DC和H₂₂-DC免疫接种后，再接种H₂₂活细胞；免疫治疗组分成P、D和HD 3小组，分别用PBS、DC和H₂₂-DC对H₂₂荷瘤小鼠进行免疫治疗。以诱发小鼠肿瘤的潜伏期大小、肿瘤生长期、肿瘤发病率以及肿瘤瘤重等为检测指标，探讨杂交瘤苗H₂₂-DC体内对H₂₂的免疫保护和治疗作用，并通过对肿瘤组织的病理学改变结合流式细胞仪检测，进一步探讨肿瘤的死亡模式。 结果1．H₂₂-DC经尾静脉注射入小鼠体内，脾、肺和肝等器官未出现肿瘤病变。2．在免疫保护组，HD组肿瘤潜伏期较P组、H组和D组明显延长（p＜0.05），肿瘤大小和肿瘤重量均明显小于P组、H组 汕头大学医学院硕士学位论文 和D组中叱刀5人3．在免疫治疗组，HD组肿瘤大小明显小于P组、 D组巾叼．05人4．肿瘤细胞流式细胞仪检测，HD组亚二倍体峰明显 高于D组和P组。 结论 1．H220C杂交瘤苗无体内致瘤性，安全可靠；2．H220C杂 交瘤苗对H。。细胞的攻击有明显的抵抗作用。H。。－DC治疗荷瘤小鼠也 有一定的作用，而以抵抗肿瘤攻击的免疫保护作用更为显著＊．HZ＊DC 杀伤肿瘤的模式为诱导H。刀瘤细胞坏死。更多还原

【Abstract】 Objective to develop vaccine by fusion of H22 hepatocarcinoma cell and DC ,and study its protective and therapeutical effect on H22 cell.Method H22-DC vaccine are produced by PEG fusion of H22 andDC induced by cytokine of spleen mononuclear cell ,sorted by CDllcmagnetic microbead marker.to study the H22-DConcogenesis,pathologically check the organs of liver,spleen and lung afterH22-DC injection from tail vein; for studying H22-DC therapeutical andprotective effect on tumor H22, two groups are divided : immune group andtherapeutic group.immune group are divided to P,H,D and HD subgroups,immunized by PBS,inactivated H22,DC and H22-DC respectively,andattacked by H22 cell, the tumor size,tumor weight , mouse survival periodand tumor latent period are recorded and statisticallyanalysized ;therapeutical group is divided into three subgroups of P,D andHD, and attacked by H22,then therapied by PBS,DC,and H22-DCrespectively. Pathological method and flow cytometry are also applied tostudy the H22JDC vaccine how to kill the H22 cell.Result 1. No oncogenesis in spleen,lung and liver after H22-DC injection; 2.1n immune group, latent period is longer in HD subgroup than in P,H and D subgroup; and tumor size and tumor weight are smaller in HD subgroup than in P,H and D subgroup; 3.In therapeutic group, tumor size is smaller in HD subgroup than in P,D subgroup.4. Flow cytometry shows higher peak of sub-diploid cells in HD than in other subgroups.Conclusion 1. H22-DC vaccine is safe without oncogenesis in vivo;2. H22-DC vaccine has distinctive protective effect on tumor H22 and can inhabit the tumor growth . 3. H22-DC can induce necrosis of H22 cell.更多还原