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

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、Western blot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mHSPs＋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。

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

Objective To investigate the antitumor immunity induced by tumor derived mixed heat shock protein/peptide(mHSPs),interleukin-12(IL-12)and Cyclophosphamide(Cy).MethodsPurified 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.ResultsThe proliferation of cytotoxic T lymphocyte(CTL)cocultured 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.ConclusionTumor derived mixed HSPs can induce powerful antitumor immune efficacy and show favorable therapeutic efficacy.