肿瘤细胞冻融裂解物上清对凋亡细胞负载的树突状细胞生物学特性的作用研究

Biological characteristics of tumor apoptotic cells-plused-dendritic cells induced by tumor lysates supernatants

目的 :探讨Balb/c小鼠骨髓瘤SP2 / 0细胞冻融裂解物上清对SP2 / 0凋亡细胞负载的骨髓来源的树突状细胞 (DC)的成熟、生物学特性及功能的影响。方法 :采用小鼠重组粒细胞 -巨噬细胞集落刺激因子 (GM -CSF)和白介素 4(IL - 4)诱导骨髓来源的DC ,10 0Gyγ射线辐照诱导SP2 / 0细胞凋亡 ,与未成熟DC共育 16- 2 0h ;SP2 / 0细胞冻融裂解物上清和脂多糖 (LPS)分别作用于DC 48h。采用免疫荧光标记和流式细胞术分析DC表型 ;[3H] TdR掺入试验和 [51Cr]释放试验测定DC刺激T细胞增殖和细胞杀伤效应。采用腹股沟皮下DC主动免疫治疗SP2 / 0荷瘤小鼠 ,每间隔 14d治疗 1次 ,共 3个疗程 ,观察肿瘤生长情况及小鼠生存期。结果 :(1)经SP2 / 0冻融裂解物上清和脂多糖 (LPS)诱导凋亡肿瘤细胞负载的DC高表达共刺激分子CD40、CD80、CD86和MHC -Ⅱ分子 ,但抗原摄取能力均下降 ;(2 )SP2 / 0冻融裂解物上清组和LPS组DC体外刺激T细胞增殖和激活CTL能力均高于对照组 (P <0 0 5 ) ;(3 )经SP2 / 0冻融裂解物上清组主动免疫治疗小鼠存活期长于其它各组 (P <0 0 5 )。结论 :小鼠SP2 / 0细胞冻融裂解物上清可诱导凋亡细胞负载的树突状细胞成熟 ,能够刺激T细胞增殖和激发肿瘤特异性细胞毒性T淋巴细胞(CTL) 。

AIM: To prepare the efficient tumor-DC vaccines, dendritic cells(DC) derived from 6-8 weeks Balb/c mice bone marrow progenitor cells were pulsed by apoptotic SP2/0 tumor cells and induced maturation by SP2/0 tumor lysates supernatants. Then SP2/0 tumor burdening Balb/c mice were immunized by the tumor-DC vaccines to observe the therapeutic effects in vivo .METHODS: Immature DC were derived by recombinant murine GM-CSF and IL-4, then were pulsed by SP2/0 apoptotic cells. Tumor-DC vaccines were stimulated by LPS and SP2/0 tumor lysates supernatants prepared by four cycles repetitive freezing and thawing, respectively. -thymidine incorporation test and standard 4h [ 51 Cr] release assay were used to detect the proliferation and activation of cytotoxic T lymphocytes (CTL) stimulated by DC in vitro . (4-5)×10 5 DC were immunized in the right inguen of SP2/0 tumor burdening Balb/c mice and most mice received three cycles immunization every two weeks. Changes of the tumor and mice life-spans were recorded. RESULTS: In vitro proliferation and activation of CTL induced by the tumor-DC vaccines of tumor lysates supernatants or LPS stimulation group were more powerful than other groups ( P<0.05 ). According to the regression of tumor and the life-spans of tumor burdening mice, therapeutic effects of tumor lysates supernatants stimulation group were the best, compared with other groups ( P<0.05 ). CONCLUSION: Tumor-DC vaccines, pulsed by apoptotic SP2/0 tumor cells and induced maturation by tumor lysates supernatants, could activate the efficient tumor specific immunity and prolong the life-span of tumor burdening mice.