摘要
目的 研究经改造的HPV16E6/E7蛋白质疫苗 (HPV 16mE6Δ/mE7)对表达HPV16E6和E7的小鼠肺癌细胞的体内生长抑制作用 ,为肺癌免疫治疗提供新线索。方法 预防性实验 :对C5 7BL/6小鼠先进行免疫 ,然后接种表达HPV 16E6和E7癌蛋白的TC 1肿瘤细胞 ;第 3 3天 ,对未出瘤小鼠进行第二次大剂量肿瘤细胞攻击。治疗性实验 :C5 7BL/6小鼠先接种TC 1细胞 ,然后进行两次免疫治疗 ,再对未出瘤小鼠进行第二次大剂量TC 1细胞攻击实验。计算各组肿瘤发生率、肿瘤体积。采用MTT方法检测淋巴细胞增殖反应。结果 预防性实验 :在 10 0天的观察时间内 ,免疫组小鼠可以有效抵抗第一、二次肿瘤细胞攻击 ,所有小鼠均未见肿瘤发生 ;而PBS组及单纯佐剂组小鼠在第一次肿瘤细胞接种后 6~ 12天内均有肿瘤形成 ,3 0天内因肿瘤负荷出现衰竭。在治疗性实验中 ,免疫组、PBS组、单纯佐剂组小鼠成瘤率分别为 2 0 %、90 %、60 % ;第二次攻击实验中 ,免疫组仍有 87.5 %的小鼠无肿瘤生长 ,而其他两组动物在 4~ 6天内均有肿瘤形成。MTT实验表明经mE6Δ/mE7免疫的小鼠脾淋巴细胞增殖反应明显强于对照组。结论 经改造的HPV16型mE6Δ/mE7蛋白质疫苗对于实验动物肺癌有较好的抑制作用 。
Objective To investigate the outgrowth inhibition of the HPV16-positive murine lung tumor induced by a modified HPV16 mE6Δ/mE7 recombinant fusion protein vaccine in vivo and provide a new clue for the further immunotherapy. Methods For prophylactic experiments, C57BL/6 mice were immunized with mE6Δ/mE7 fusion protein, and then inoculated with the TC-1 tumor cell, expressing HPV16 E6 and E7 viral proteins. On day 33 after inoculation, the tumor-free mice were re-challenged with a larger dose of TC-1 tumor cells. For therapeutic experiments, mice were vaccinated with mE6Δ/mE7 on days 3 and 14 after tumor cell inoculation. On day 60, the tumor-free mice were re-challenged with a larger dose of tumor cells. Tumor incidence and tumor volume of each group were calculated. MTT method was used to determine the proliferation of lymphocyte. Results In the prophylactic experiments, immunization with the mE6Δ/mE7 completely protected the mice against the tumor cell challenge and rechallenge, and all the mice remained tumor free during the 100 days’ observation period. In contrast, all the mice in PBS and IFA-treated groups developed tumors within 6--12 days after the first tumor cell inoculation, and died of tumor burden within 30 days. In the therapeutic experiments, the tumor formation rates were 20%, 90% and 60% in vaccinated, PBS and IFA groups respectively. In the next larger dose of tumor cells rechallenge experiment, 87.5% of vaccinated mice still remained tumor free, but all the mice from either PBS or IFA group developed tumors with 4--6 days. In addition, the results of MTT indicated that the proliferation of lymphocytes from vaccinated mice was stronger than that from control group. Conclusion The modified mE6Δ/mE7 can efficiently inhibit the growth of lung cancer in the animal model, indicating that mE6Δ/mE7 protein-based vaccine might show promise for the future clinical application.
出处
《中国肺癌杂志》
CAS
2004年第5期379-382,共4页
Chinese Journal of Lung Cancer
基金
国家自然科学基金 (No .39970 81 8和No.30 1 71 0 58)
"863"项目 (2 0 0 2AA2 1 60 4 1 )资助~~