肿瘤细胞裂解物联合IL-2预防黑色素瘤发生并抑制肿瘤生长的免疫机制

目的 探讨肿瘤细胞裂解物（TCL）联合IL-2对黑色素瘤的预防和抑制作用及其免疫机制。方法 超声破碎仪制备B16F10黑色素瘤细胞TCL。24只C57BL/6小鼠随机分为四组，分别予PBS、IL-2、TCL及TCL+IL-2预防性免疫3周，第4周对侧植瘤。观察小鼠出瘤时间及肿瘤大小，连续采集外周血行流式细胞术动态监测CD4+T及CD8+T细胞。流式细胞术及免疫组织化学检测小鼠脾脏及肿瘤组织CD4+T及CD8+T细胞。结果 TCL+IL-2组预防性免疫后明显延迟了小鼠的出瘤时间（P=0.034），且肿瘤体积（P=0.023）及瘤重（P=0.0015）也明显小于对照组。流式细胞术动态监测结果提示TCL+IL-2组及单纯TCL组外周血中CD8+T细胞较对照组均有明显上升（P=0.0016, P=0.012）。TCL+IL-2组的CD4+T细胞在植瘤后较对照组有所下降（P=0.0089）。脾脏及肿瘤组织中CD4+T细胞及CD8+T细胞的结果与外周血中的表达趋势相同。结论 TCL联合IL-2的肿瘤疫苗通过激活CD8+T细胞途径，预防小鼠黑色素瘤的发生并抑制肿瘤生长。

Immune Mechanism of Tumor Cell Lysate Combined with IL-2 Preventing Melanoma and Inhibiting Tumor Growth

Objective To investigate the preventive and inhibitory effects of tumor cell lysate(TCL) combined with IL-2 on melanoma and the potential immune mechanism. Methods The B16F10 melanoma TCL cell were prepared using an ultrasonic disruptor. Twenty-four C57BL/6 mice were randomly divided into four groups which were immunized with PBS, IL-2, TCL and TCL+IL-2 for three weeks, and contra lateral tumors were implanted in the fourth week. We observed onset time of tumor and tumor size, collected peripheral blood continuously and monitored the expression of CD4+T and CD8+T cell subsets dynamically by flow cytometry. Spleen and tumor tissues of mice were also tested for CD4+T and CD8+T cell subsets by flow cytometry and immunohistochemistry, respectively. Results The preventive immunization of the TCL+IL-2 group significantly delayed the onset time of tumor (P=0.034); moreover, the tumor volume (P=0.023) and tumor weight (P=0.0015) were also significantly smaller than those in the control group. The expression of CD8+T cell subsets in the TCL+IL-2 group and the TCL-only group were significantly higher than that in the control group (P=0.0016, P=0.012). However, the CD4+T cell subsets of the TCL+IL-2 group decreased after tumor implantation, compared with the control group (P=0.0089). The expression of CD4+T and CD8+T cell subsets in spleen and tumor tissues were as same as those in peripheral blood. Conclusion The tumor vaccine of TCL combined with IL-2 could prevent the occurrence of melanoma in mouse and effectively inhibit tumor growth by activating CD8+T cells.