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目的探讨黑素瘤肺转移小鼠模型体内表达可溶性PD-1(sPD-1)对B7-H1/PD-1信号传导的阻断作用,及联合Hsp70-B16抗原肽对小鼠黑素瘤肺转移的抑制作用。方法建立小鼠黑素瘤肺转移模型,应用免疫组织化学染色和流式细胞术检测肺转移灶PD-1及B7-H1的表达情况,从小鼠尾静脉注射sPD-1表达质粒,并联合应用Hsp70-B16抗原肽皮下免疫小鼠,于接种瘤细胞后第17天解剖小鼠,观察黑素瘤肺转移的情况,检测各组小鼠肺转移灶局部淋巴细胞浸润及部分免疫学指标。结果小鼠黑素瘤肺转移模型成功建立,肺转移灶局部有大量的PD-1阳性细胞,转移瘤B16细胞表面表达较多B7- H1分子,静脉注射sPD-1表达质粒联合抗原肽免疫治疗组小鼠肺转移明显受抑制,抑制率达95%,显著高于对照组,其对应组小鼠肺部CD8~+T淋巴细胞的数量、脾淋巴细胞的细胞毒活性及血清中的IL-2和IFN-γ的浓度均明显高于对照组(P<0.01)。结论小鼠体内转染表达sPD-1可阻断B7-H1/PD-1信号传导,抑制小鼠黑素瘤肺转移,联合应用Hsp70-B16抗原肽具有明显的协同作用。
Objective To investigate the blocking effect of soluble PD-1 (sPD-1) on B7-H1 / PD-1 signal transduction in melanoma lung metastasis mouse model and the effect of Hsp70-B16 antigen peptide on melanoma lung Inhibition of metastasis. Methods The mouse model of melanoma lung metastasis was established. The expression of PD-1 and B7-H1 in lung metastasis was detected by immunohistochemical staining and flow cytometry. The sPD-1 expression plasmid was injected from the tail vein of mice and combined with the application Mice were immunized subcutaneously with Hsp70-B16 antigen peptide, and the mice were dissected on the 17th day after inoculation of the tumor cells. The metastasis of melanoma was observed. The local lymphocyte infiltration and some immunological indexes of lung metastases were detected. Results The mouse melanoma lung metastasis model was successfully established. There were a large number of PD-1 positive cells in the lung metastases, more B7-H1 molecules on the surface of the metastatic B16 cells, intravenous injection of the sPD-1 expression plasmid and antigen peptide immunotherapy The lung metastasis of mice was significantly inhibited, the inhibition rate was 95%, which was significantly higher than that of the control group. The number of CD8 ~ + T lymphocytes, the cytotoxic activity of splenic lymphocytes and IL- 2 and IFN-γ concentrations were significantly higher than the control group (P <0.01). Conclusion The expression of sPD-1 can block the B7-H1 / PD-1 signal transduction and inhibit the metastasis of melanoma in mice. The combined application of Hsp70-B16 antigen peptide has obvious synergistic effect.