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目的探讨光动力疗法(photodynamic therapy,PDT)联合瘤体内输注树突状细胞(dendritic cell,DC)的光免疫疗法(photody-namic immuno-therapy,PIT)对小鼠Heps肝癌移植瘤的抑制作用及免疫效应。方法体外培养昆明小鼠骨髓源性DC,4,’6-二脒基-2-苯基吲哚(4,6-diamidino-2-phenylindole,DAPI)荧光染色液标记DC备用。128只昆明小鼠皮下接种Heps肝癌细胞建立肿瘤模型,随机分为对照组、PDT组、DC组和PIT组。对照组小鼠瘤体内注射生理盐水,PDT组单纯PDT治疗,DC组小鼠瘤体内注射DAPI标记的DC,PIT组PDT联合瘤体内注射DAPI标记的DC细胞。治疗后定期测量各组肿瘤体积,记录各组小鼠生存时间,荧光显微镜下计数DC组及PIT组小鼠淋巴结中荧光细胞数目,流式细胞仪测定各组小鼠外周血T细胞亚群,LDH释放法测定各组小鼠脾细胞杀伤活性。结果 (1)与对照组相比,PDT组与PIT组治疗后肿瘤生长明显受抑;(2)PDT组与PIT组小鼠生存时间延长;(3)高倍镜视野下DC组较PIT组荧光细胞数增多(P<0.05);(4)治疗后72 h,PDT及PIT组小鼠外周血CD8+T细胞百分率均明显高于对照组和DC组(P<0.01、P<0.01),其中PIT组较PDT组增高明显(P<0.01),(5)PDT组与PIT组小鼠脾脏细胞杀伤活性较对照组和DC组明显增强(P<0.01,P<0.01)。结论 PDT疗法能够抑制肿瘤生长并激发宿主免疫应答,联合输注DC可增强PDT对小鼠Heps肝癌移植瘤的抑制作用及免疫效应。
Objective To investigate the inhibitory effect of photodynamic therapy (PDT) combined with photody-namic immuno-therapy (PIT) on transplanted Heps hepatocellular carcinoma in vivo And immune effect. Methods Kunming mouse bone marrow-derived DCs were cultured in vitro, and labeled with 4,6-diamidino-2-phenylindole (DAPI). 128 Kunming mice were inoculated subcutaneously with Heps liver cancer cells to establish a tumor model and were randomly divided into control group, PDT group, DC group and PIT group. The control mice were injected with normal saline and PDT group were treated with PDT alone. DAPI-labeled DCs were injected into the DCs in vivo and DAPI-labeled DCs in the PIT group were injected with PDT. After treatment, the tumor volume of each group was measured regularly. The survival time of each group was recorded. Fluorescent microscope was used to count the number of fluorescent cells in lymph nodes of DC group and PIT group. The T lymphocyte subsets in peripheral blood were measured by flow cytometry. LDH release assay was used to determine the killing activity of spleen cells in each group. Results (1) Compared with the control group, the tumor growth of PDT group and PIT group was obviously inhibited; (2) The survival time of mice in PDT group and PIT group was prolonged; (3) The fluorescence intensity of DC group was higher than that of PIT group (P <0.05). (4) At 72 h after treatment, the percentage of CD8 + T cells in PDT and PIT groups was significantly higher than that in control group and DC group (P <0.01, P <0.01) PIT group was significantly higher than PDT group (P <0.01). (5) The killing activity of spleen cells in PDT group and PIT group was significantly higher than that in control group and DC group (P <0.01, P <0.01). Conclusions PDT can inhibit tumor growth and stimulate host immune response. Combined infusion of DC can enhance the inhibitory effect and immune effect of PDT on Heps hepatocellular carcinoma in mice.