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

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

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。

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.