半抗原DNP修饰自体肿瘤疫苗治疗晚期恶性黑色素瘤的临床研究

摘要

目的：探讨半抗原二硝基氟苯（dinitrophenyl，DNP）修饰自体肿瘤疫苗在晚期恶性黑色素瘤治疗中的作用。方法84例Ⅲ期或Ⅳ期可切除恶性黑色素瘤患者随机分为半抗原DNP修饰瘤苗组和半抗原DNP未修饰瘤苗组，每组42例，分别采用半抗原DNP修饰的自体肿瘤疫苗和半抗原DNP未修饰的自体瘤苗联合化疗进行治疗，采用流式细胞技术检测两组患者外周血CD4+CD25+调节性T细胞（regulatory T cell，Treg）数量及淋巴细胞亚群的变化，观察迟发型超敏反应（delayed type hypersensitivity，DTH）及随访生存情况。结果治疗后半抗原DNP修饰瘤苗组患者CD4+-IFN-γPE、CD8+-IFN-γPE淋巴细胞亚群较治疗前及半抗原DNP未修饰瘤苗组明显升高（P均<0.05），而CD4+CD25+Treg数量减少（P<0.05），半抗原DNP未修饰瘤苗组治疗前后无明显变化（P>0.05）；半抗原DNP修饰瘤苗组患者较半抗原DNP未修饰瘤苗组DTH明显增强，硬结明显增大，两组DTH达到阳性结果（≥5 mm）的百分比分别为94.6%和45.0%（P<0.05）。半抗原DNP修饰瘤苗组患者1年、2年和3年生存率分别为95.0%、73.0%和65.5%，总生存率（overall survival，OS）为79.3%，无疾病生存率（disease free survival，DFS）为81.1%；半抗原DNP未修饰瘤苗组患者1年、2年和3年生存率分别为90.1%、59.0%和43.5%，OS为64.2%，DFS为74.0%，两组患者生存时间差异有统计学意义（P<0.05）。结论半抗原DNP修饰的自体瘤苗可增强恶性黑色素瘤患者特异性细胞介导的免疫反应，抑制机体免疫耐受，从而延长患者生存时间。%Objective To investigate the efficacy of autologous tumor vaccine modified to carry the dinitrophenyl hapten to treat advanced malignant melanoma. Methods Clinical data were retrospectively analyzed for 84 patients who underwent resection for stageⅢb,Ⅲc or IV malignant melanomas. Patients were divided into two groups depending on whether or not their biochemotherapy was supplemented with dinitrophenyl-modified autologous tumor vaccine. The two groups were compared in terms of the following parameters:relative numbers of CD4+CD25+ regulatory T cells and lymphocyte subsets in peripheral blood,which were determined using flow cytometry;observed strength of delayed type hypersensitivity(DTH);and clinical outcomes such as overall survival(OS)and disease-free survival (DFS). Results The subpopulations of CD4+-IFN-γPE and CD8+-IFN-γPE lymphocytes were significantly larger in the vaccinated patients than in the biochemotherapy-only patients (P<0.05),while the relative proportions of CD4+CD25+ regulatory T cells were significantly smaller(P<0.05). DTH-positive response(defined as spot size ≥5 mm)occurred in 94.6% of vaccinated patients but in only 45% of biochemotherapy-only patients (P<0.05). The 1-,2- and 3-year survival rate were 95.0%,73.0 and 65.5% in the vaccinated group,respectively,90.1%,59.0%and 43.5% in the biochemotherapy-only group,respectively. OS and DFS in the vaccinated group were 79.3% and 81.1%,compared to 64.2% and 81.1% in the biochemotherapy-only group. The difference in OS between the two groups was significant (P<0.05). Conclusion Administering autologous tumor vaccine modified with the dinitrophenyl hapten may increase the overall survival of patients with specific cell-mediated melanoma,and it appears to do so by en-hancing their immune response and inhibiting immune tolerance.