中华皮肤科杂志

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可溶性PD-1及联合Hsp70-B16抗原肽复合物对小鼠黑素瘤肺转移的抑制作用

邱惠1, 张桂梅2, 张慧2, 耿辉2, 冯作化2   

  1. 1. 武汉大学中南医院放化疗科, 430071;
    2. 华中科技大学同济医学院生物化学与分子生物学教研室
  • 收稿日期:2006-12-19 出版日期:2007-11-15 发布日期:2007-11-15
  • 基金资助:
    国家自然科学基金(30600735)

Inhibitory effect on pulmonary metastasis of melanoma in mice by soluble PD-1 and its combina-tion with HspT0-B16 antigen peptides

QIU Hui1, ZHANG Gui-mei2, ZHANG Hui2, GENG Hui2, FENG Zuo-hua2   

  1. Department of Radiotherapy and Chemotherapy, Zhongnan Hospital, Wuhan University, Wuhan 430071, China
  • Received:2006-12-19 Online:2007-11-15 Published:2007-11-15

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摘要： 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.

关键词: 黑色素瘤, 实验性, 基因疗法, HSP70热休克蛋白质类, sPD-1

Abstract: Objective To investigate the blockade effects of soluble PD-1 (sPD-1) expressed in vivo on B7-H1/PD-1 signal transduction,and inhibitory effect in pulmonary metastasis of melanoma with combination of Hsp70-B16 antigen peptides in mice.Methods The pulmonary metastasis model of melanoma was established in mice.Immunohistochemical staining and flow cytometry were utilized to detect the expression of PD-1 and B7-H1 respectively in pulmonary metastasis loci.Four days after the inoculation of tumor cells,forty murine models of pulmonary metastasis were randomly divided to be immunized with normal sodium (group A),empty vector pcDNA3.1 (group B),PDlA plasmid (group C) respectively via tail vein injection,subcutaneous injection of Hsp70-B16 antigen peptides (group D) or with the combination of intravenous PDlA plasmid and subcutaneous Hsp70-B 16 antigen peptides (group E).The local infiltration with lymphocytes in pulmonary metastasis loci was observed and a series of immunological parameters were assessed 17 days after the inoculation of tumor cells.Results The melanoma pulmonary metastasis model was successfully established.There were a lot of PD-1 positive cells in these loci,and B7-H1 molecule was massively expressed on the surface of B16 cells in metastasis loci.The pulmonary metastasis was inhibited in the mice of group E,and the inhibition rate was 95%,higher than that in other groups (53%,76%,9% in group C,D,B,respectively).The quantity of CD8+ T cells in pulmonary metastasis loci,cytotoxicity of spleen lymphocytes to tumor cells,and serum concentration of IL-2 and IFN-γwere all significantly elevated in the mice of group E as compared with those of other groups (all P<0.01).Conclusions The expressed sPD-1 in vivo can block the signal transduction between B7-H1 and PD-1,and inhibit the metastasis of melanoma in mice.Also,the combined Hsp70-B16 peptides could enhance the effect of sPD-1.

Key words: Melanoma,experimental, Gene therapy, HSP70 heat-shock proteins, sPD-1