人乳头状瘤病毒16型E6基因转染人外周血树突状细胞的体内抗宫颈癌研究

Antitumor Effect Induced by Dendritic Cell from Human Peripheral Blood Transfected with Human Papillomavirus Type 16 E6 Antigen Gene in Mice

目的:观察人乳头状瘤病毒(HPV)16型E6基因转染人外周血来源的树突状细胞(DC)制备的疫苗,在严重联合免疫缺陷(SCID)小鼠体内对人宫颈癌(CaSki)细胞的免疫治疗和免疫保护作用。方法:细胞因子扩增人外周血DC,脂质体(Li-pofectamine)转染HPV16E6制备HPV16E6-DC疫苗,进行致瘤性检测,并观察在SCID小鼠体内疫苗抗人CaSki细胞的免疫保护及对荷瘤小鼠的免疫治疗作用。结果:①HPV16E6-DC疫苗接种于小鼠体内未见肿瘤形成,脾、肺、肝和肾等器官未出现肿瘤病变。②在免疫治疗组,用HPV16E6-DC疫苗治疗的荷瘤小鼠,肿瘤生长速度较对照组慢,瘤体积小于对照组,瘤重量较对照组明显减轻(P<0.01)。③在免疫保护组,经HPV16E6-DC疫苗免疫的小鼠,CaSki细胞攻击后,肿瘤潜伏期较对照组明显延长(P<0.01),瘤体积明显小于对照组,瘤重量也轻于对照组(P<0.01)。结论:人外周血来源的HPV16E6-DC疫苗无体内致瘤性,且能在一定程度上抑制人CaSki细胞在SCID小鼠体内的生长,抵抗肿瘤细胞的攻击,对机体具有相应的抗肿瘤免疫治疗及保护作用。

Objective： To study the immunotherapeutic and immunopreventive effects of dendritic cell （DC）from human peripheral blood transfected with human papillomavirus（ HPV）Type 16 E6 antigen gene on CaSki cells in SCID mice. Methods： DC-enriched populations were prepared from human peripheral blood mononuclear cell with the combination of recombinant human granuloeyte-macrophage colony-stimulation factor, recombinant human interleukin-4 and recombinant human tumor necrosis factor-α. The plasmid containing HPV16 E6 gene was transfected into DC with lipofectarnine. The antitumor effects, immunotherapeutic and immunopreventive effect of HPV16 E6-DC vaccine were studied in SCID mice. Results： No tumor was found in spleen, hmg, liver and kidney after the HPV16 E6-DC vaccine injection. In immunotherapeutic group, the growth speed of tumor decreased, and the tumor volume was less in control subgroup, and tumor weight was lighter in control subroup（ P 〈 0.01 ）. In immunopreventive group, the tumor latent period was longer in HPV16 E6-DC subgroup, compared with control subgroup（ P 〈 0.01）, and the tumor volumn was less in HPV16 E6- DC subgroup, and tumor weight was lighter in HPV16 E6-DC subgroup（ P 〈 0.01）. Conclusion： Human peripheral blood derived HPV16 E6-DC vaccine has no oncogenesis in vivo, and can inhibit the growth of CaSki cells in SCID mice and protect mice from attack by CaSki ceils, which indicates its immtmotherapeutic and immtmopreventive effects on tumor.