裸鼠肝包膜下输注DC-CIK细胞抑制脾内肿瘤和肝转移瘤生长的初步研究

摘要

Objective To develop a method for injecting immune cells that target the liver and suppress tumor growth there. Method A hollow fiber pad was placed into the liver subcapsule of nu-mice bearing SW480 tumors,leading to cyst formation in the subcap-sule. Dendritic cells(DCs)and cytokine-induced killer cells(CIKs)were induced from human peripheral blood monocytes. The ability of CIKs to inhibit SW480 cell proliferation was measured. DCs were loaded with antigens from SW480 cells,and co-cultured with CIKs. DC-CIK co-cultures were injected into the subcapsule cyst or into the caudal vein(6×107 cells in 0.3 mL),while the same vol-ume of saline was injected into the cyst or caudal vein in control groups. Each group received injections twice a week for 4 weeks. At 48 h after the last injection,growth inhibition of the primary tumors in spleen and mean numbers of metastic tumors in liver were determined. Results Mixing CIKs and SW480 cells in the ratio of 60:1 inhibited SW480 proliferation by (75.3±8.42)%. CD3/CD56 were expressed on (36.4±13.2)%of CIK cells. Tumor inhibition was 32.07%in the animals injected in the cyst and 16.54%in the ani-mals injected in the caudal vein(P<0.05);mean numbers of metastic liver tumors were 2.47±1.02 in the group injected in the cyst and 5.05±1.06 in the group injected in the caudal vein (P<0.01). Conclusion Injecting DC-CIKs into nu-mice may significantly inhibit the growth of primary tumors in spleen and metastic tumors in liver,and the effects are greater when the cells are injected into the liver subcapsule than into the caudal vein.%目的：建立一种肝脏靶向输注免疫效应细胞法，用于抑制脾内肿瘤和肝转移瘤生长。方法将一空心纤维垫置于接种SW480肿瘤模型裸鼠肝包膜下，形成肝包膜下人工囊腔；从人外周血单核细胞诱导出细胞因子诱导杀伤细胞（cytokine induced killer，CIK）和树突状细胞（dendritic cell，DC）。体外检测CIK对SW480细胞杀伤率，并使DC负载SW480细胞抗原。将共同培养的效应细胞（DC-CIK）注入模型裸鼠肝包膜下人工囊腔（A组，20只）和尾静脉（C组，20只），每只6×107个/0.3 ml；同法对20只模型裸鼠分别注入生理盐水，0.3 ml/只，并分为B组、D组各10只（作为A组、C组的对照）。每周注射2次，共注射4周。末次注射后48 h，检查比较A组和C组脾内原发肿瘤体积、抑瘤率和肝组织病理切片转移瘤灶均数。结果效靶比为60：1时CIK对SW480细胞杀伤率为（75.3±8.42）%。CIK的CD3/CD56表达率为（36.4±13.2）%。治疗后，A组和C组的抑瘤率分别为32.07%和16.54%（P<0.05）；A组和C组肝组织切片转移瘤灶均数分别为（2.47±1.02）个和（5.05±1.06）个（P<0.01）。结论裸鼠肝包膜下输注DC-CIK可明显抑制脾内肿瘤和肝转移瘤生长，抑瘤效果优于尾静脉注射DC-CIK。