目的观察混合热休克蛋白/肽(mHSP/P)疫苗、环磷酰胺(Cy)及重组人IL-2(rhIL-2)联合免疫治疗小鼠肉瘤的效果。方法取小鼠S180细胞肉瘤组织,经层析分离获得混合mHSP/P疫苗,进行常规蛋白质印迹鉴定。S180细胞接种至小鼠皮下后第2天开始实验,将小鼠按mHSP/P、Cy、rhIL-2药物组合、疗程和剂量的不同依次分为B～I组,以及给予生理盐水的A组(对照组)。自肿瘤移植后至第45天,观察计算肿瘤体积;以肿瘤移植后3个月仍无瘤生长或肿瘤逐渐缩小至消失为标准,观察各组小鼠的无瘤生存情况,并进行平均生存期比较;以对照组的肿瘤体积为参照,计算第35天各组小鼠肿瘤的生长抑制率。结果蛋白质印迹分析结果显示所获目的蛋白为含有HSP110、HSP70、HSP60、Gp96的混合mHSP疫苗。3种药物联合应用的治疗效果明显优于单用mHSP/P疫苗,且多疗程较单一疗程疗效好,其中rhIL-2的最佳治疗剂量为5×104～1×105IU/只,并可使30%的小鼠肿瘤消退而长期生存(治愈),平均生存期延长22～26d,肿瘤生长抑制率为65%～76%。结论mHSP疫苗与适量rhIL-2以及小剂量Cy联合应用为临床肉瘤治疗新的免疫治疗方案提供了实验基础。 Objective To observe the effect of combined heat shock protein/peptide (mHSP/P) vaccine, cyclophosphamide (Cy) and recombinant human IL-2 (rhIL-2) on immunotherapy of mouse sarcoma. Methods The mouse S180 cell sarcoma tissue was obtained and the mixed mHSP/P vaccine was obtained by chromatographic separation for routine Western blot identification. After S180 cells were inoculated into mice, they were started on the second day after subcutaneous injection. The mice were divided into groups B to I according to mHSP/P, Cy, rhIL-2 drug combinations, treatment duration, and dose, and group A given physiological saline. (control group). From the time of tumor transplantation to the 45th day, the tumor volume was observed and calculated; the tumor-free survival of the mice in each group was observed and the average survival time was observed after tumor transplantation was continued for 3 months after tumor transplantation or the tumor was gradually reduced to disappear. For comparison, the growth inhibition rate of mice in each group on the 35th day was calculated using the tumor volume in the control group as a reference. Results Western blot analysis showed that the obtained protein was a mixed mHSP vaccine containing HSP110, HSP70, HSP60, and Gp96. The therapeutic effect of the combined use of the three drugs was significantly better than that of the mHSP/P vaccine alone, and multiple courses of treatment were more effective than the single course of treatment. The optimal therapeutic dose of rhIL-2 was 5×104 to 1×105 IU/body and it could be used. In 30% of the mice, the tumor regressed and survived for a long time (cure). The average survival time was 22 to 26 days, and the tumor growth inhibition rate was 65% to 76%. Conclusion The combination of mHSP vaccine, appropriate amount of rhIL-2 and low-dose Cy can provide experimental basis for clinical sarcoma treatment of new immunotherapy options.