结核杆菌抗原85A与GM-CSF诱发抗鼠黑色素瘤效应研究

Study of anti-melanoma effect in mice injected with melanoma cells transfected with the recombinant expressing Ag85A and GM-CSF

目的:用结核杆菌抗原85A(Ag85A)和细胞因子GM-CSF,作为异种抗原及免疫增强剂,研究其在鼠体内能否有效地诱导抗黑色素瘤免疫应答。方法:用分子生物学法构建重组质粒pRSC-Ag85A/GM-CSF,通过细胞转染获得稳定表达Ag85A和GM-CSF的B16黑色素瘤细胞。48只C57BL/6小鼠随机均分为B16细胞组、B16-pRSC细胞组、B16-Ag85A细胞组和B16-Ag85A/GM-CSF细胞组,分别将上述不同B16细胞经背部皮下接种小鼠。通过观察其致瘤性的强弱、荷瘤鼠生存时间、鼠血清IFN-γ水平以及CTL、NK细胞杀伤活性,分析Ag85A和GM-CSF在抗肿瘤免疫效应中作用。结果:B16-Ag85A/GM-CSF细胞组小鼠肿瘤生长明显受到抑制,生存时间较对照组平均延长40%,血清IFN-γ分泌较对照组高1倍,CTL、NK细胞杀伤活性与其他各组相比也有较大的提高。结论:接种稳定表达Ag85A和GM-CSF的B16细胞能降低对小鼠的致瘤性,诱导有效的抗肿瘤效应。

Objective：To study anti-melanoma effect in mice injected with melanoma cells expressing Mycobacterium tuberculosis antigen 85A（Ag85A） and GM-CSF as heterogeneity antigen and immunoadjuvants.Methods：The recombinant pRSC-Ag85A/GM-CSF was constructed by the molecular biological technology and was transfected into the melanoma B16 cells and the transcription of Ag85A and GM-CSF in B16 cells was respectively detected by RT-PCR.The different gene-modified B16 cells were injected into C57BL/6 mice and the tumorigenicity was analyzed by measuring the size of tumor and observing the survival of tumor-bearing mice.Meanwhile,the anti-tumor immune response induced by the Ag85A and GM-CSF was evaluated by detecting the activities of CTL and NK cells,and serum cytokine level respectively.Results：Mice injected with B16 cells transfected with the recombinant pRSC-Ag85A/GM-CSF could survive longer than the other mice.Compared with the control group,the level of IFN-γ in the serum and the activities of CTL and NK cells were obviously increased and the difference was statistically significant.Conclusion：B16 cells transfected with the recombinant pRSC-Ag85A/GM-CSF could reduce the tumorigenicity and effectively induce the anti-tumor effect in mouse model.