混合性热休克蛋白/肽疫苗与白细胞介素-12联合诱导特异性细胞毒T淋巴细胞产生的实验研究

Combined Therapy with Mixed Heat Shock Protein/Peptide Vaccine and Interleukin-12 Results in Enhanced Antitumor Effects

目的研究肿瘤组织来源混合热休克蛋白(mHSPs)/肽疫苗诱导产生特异性细胞毒性T淋巴细胞(CTL)的作用及其抗肿瘤免疫效应,为应用其治疗人类恶性肿瘤提供实验依据。方法利用细胞培养、流式细胞技术、蛋白质的分离纯化技术、电泳技术、Westernblot检测方法、T淋巴细胞的诱导及体外扩增、乳酸脱氢酶法等,测定肿瘤多肽复合物诱导活化小鼠体内CTL杀伤效应。结果mHSPS免疫组和mHSPs+Cy+IL-12免疫组体外诱生的CTL反应活性均比对照组的CTL活性有所升高,尤其mH-SPs+Cy+IL-12免疫组升高明显,且随效靶比的上升而上升;与正常小鼠(CD4+46%、CD8+18%)比较,对照组、mHSPs免疫组及mHSPs+Cy+IL-12免疫组小鼠CD4+亚群的比例分别为38%、46%、54%。CD8+亚群的比例分别为26%、22%、16%。结论混合热休克蛋白/肽可诱发强大的抗肿瘤免疫效应,当与IL-12、低剂量Cy者联合应用后免疫功能增强最为明显。

Objective To investigate the antitumor immunity induced by tumor derived mixed heat shock protein/peptide （mHSPs）, interleukin-12 （IL-12） and Cyclophosphamide （Cy）. Methods Purified mixed HSP was prepared from tumor by S180 protein extraction and purification, SDS-PAGE, Western blot and animal experiment were applied for mixed HSPs analysis. Results The proliferation of cytotoxic T lymphocyte （CTL） cocuhured in the mHSPs＋ Cy＋ IL-12 group was significantly remarkable and the content of CD8^＋ CTLs was significantly more in comparison with the other groups （P〈0.01）. To the tumor bearing mice, mHSPs ＋Cy＋IL-12 group showed partial therapeutic efficacy, the averaged survival period was over 60 d, and 90% of the mice in this group got long period tumor free survival （〉90 d）, obvious difference （P〈0.05） from the other groups. Conclusion Tumor derived mixed HSPs can induce powerful antitumor immune efficacy and show favorable therapeutic efficacy.