进展期肿瘤主动特异性免疫治疗的临床研究

目的探讨自体肿瘤疫苗的作用机制。方法50例进展期肿瘤患者术后第4周开始以自体肿瘤细胞疫苗行主动特异性免疫治疗。免疫接种共4次,每次间隔7-10 d;接种前3 d及第4次接种后1周,采集外周血,分离单个核细胞,以流式细胞分析术/细胞内细胞因子检测法测定CD8+-IFN-γ+、CD8+-IL-10+细胞及CD4+-INF-γ+、CD4+-IL-10+细胞;用自体肿瘤抗原做皮肤迟发型过敏试验(DTH),48 h后测量红斑、硬结大小(mm)。对DTH反应部位皮肤活检,用免疫组化染色了解局部细胞免疫反应。结果自体肿瘤细胞疫苗治疗后表现为:①CD8+-IFN-γ+阳性细胞由(3.90±1.42)%升至(7.12±2.15)%;CD4+-INF-γ+阳性细胞由(4.19±1.50)%升至(7.20±2.29)%(P<0.05);②与治疗前相比,治疗后患者对自体肿瘤抗原的特异性DTH反应明显增强(P<0.01);③DTH反应部位CD8+T细胞、CD4+T细胞浸润明显增多;④患者耐受性良好。结论自体肿瘤细胞疫苗免疫后,可改善肿瘤患者细胞介导的抗瘤免疫反应;自体肿瘤疫苗可激发患者特异性细胞介导的免疫反应。

Clinical study of active specific immunotherapy in advanced tumors

Objective To evaluate the immunological parameters of patients with resected advanced tumors after active immunotherapy using a novel autologous whole tumor cell vaccine.Methods Fifty post-operative patients with advanced tumors were enrolled in this study.The vaccination was started at 4 weeks after resection to allow the patients sufficient time to recover from immunological suppression induced by surgery.The vaccination was given 4 times at a 7-10 days interval.Intracellular cytokine of CD~+_8-IFN-γ~+,CD~+_4-INF-γ~+,CD~+_4-IL-10~+ and CD~+_8-IL-10~+ T cell in PBMNC were detected by flow cytometry 3 days before and 7 days after the final inoculation;diameters of erythemas or induratinons were observed within 48 hours after the skin tests with autologous tumor antigen.In order to assay DTH local cellular immune response,the biopsy specimens of DTH skin reactinons were analyzed by immunohistochemistry.Results ① CD~+_8-IFN-γ~+ T cell were increased from(3.90±1.42)% to(7.12±2.15)%;CD~+_4-INF-γ~+ T cell were increased from(4.19±1.50)% to(7.20±2.29)%((P<0.05)).② Specific DTH reactions to autologous tumor antigen were obviously enhanced(P<0.01).③ The dense infiltratinon with CD~+_8 T cell and CD~+_4 T cell were noticed at the site of DTH skin reaction.④ All patients showed good tolerance.Conclusion The autologous tumor vaccine can induce specific cellular immune response.It can improve the host's anti-tumor immune response.