Ren Shurong, Wang Qiubo, Zhang Yanli, Lu Cuixiu, Li Ping, Li Yumei
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Feb;33(2):145-50.
To investigate the therapeutic effect of Toll-like receptor 7( TLR7) agonist imiquimod combined with dendritic cell( DC)-based tumor vaccine on melanoma in mice and the potential mechanism.
Melanoma-bearing mouse models were established by subcutanous injection of B16-OVA cells into C57 BL /6 mice. DCs were isolated from mouse bone marrow and propagated in culture medium with recombinant mouse granulocyte-macrophage colony-stimulating factor( rm GM-CSF) and recombinant mouse interleukin-4( rm IL-4). DC vaccine( OVA-DC) was prepared by overnight incubation of DCs added with chicken ovalbumin. C57 BL /6 mice were separated into four groups which were treated with PBS,topical imiquimod application,OVA-DC intradermal injection and imiquimod plus OVA-DC,respectively. The tumor size was calculated by digital vernier caliper. Peripheral blood CD4+FOXP3+Tregs of the tumor-bearing mice was detected by flow cytometry. The cytotoxicity of splenic lymphocyte against B16-OVA was assessed in vitro by CCK-8 assay.
Compared with the other three groups,B16-OVA-bearing mice treated with imiquimod plus DC vaccine had the smallest tumor volume. The percentage of CD4+FOXP3+Tregs decreased significantly in the combined treated mice. The combined treatment enhanced significantly cytotoxicity of splenic lymphocytes against B16-OVA cells.
Imiquimod combined with antigen-pulsed-DC vaccine could reduce CD4+FOXP3+Treg proportion and promote anti-tumor effect in mice with melanoma
探讨Toll样受体7（TLR7）激动剂咪喹莫特联合树突状细胞（DC）肿瘤疫苗对小鼠黑色素瘤的治疗效果及潜在机制。
将B16-OVA细胞皮下注射到C57BL/6小鼠体内，建立荷黑色素瘤小鼠模型。从小鼠骨髓中分离DC，并在含有重组小鼠粒细胞-巨噬细胞集落刺激因子（rmGM-CSF）和重组小鼠白细胞介素-4（rmIL-4）的培养基中培养扩增。将添加鸡卵清蛋白的DC过夜孵育制备DC疫苗（OVA-DC）。将C57BL/6小鼠分为四组，分别给予PBS、局部应用咪喹莫特、皮内注射OVA-DC以及咪喹莫特联合OVA-DC治疗。用数字游标卡尺测量肿瘤大小。通过流式细胞术检测荷瘤小鼠外周血CD4⁺FOXP3⁺调节性T细胞（Tregs）。采用CCK-8法体外评估脾淋巴细胞对B16-OVA的细胞毒性。
与其他三组相比，咪喹莫特联合DC疫苗治疗的荷B16-OVA小鼠肿瘤体积最小。联合治疗组小鼠CD4⁺FOXP3⁺Tregs百分比显著降低。联合治疗显著增强了脾淋巴细胞对B16-OVA细胞的细胞毒性。
咪喹莫特联合抗原脉冲DC疫苗可降低小鼠黑色素瘤模型中CD4⁺FOXP3⁺Treg比例，增强抗肿瘤作用
