CAJ | 학술논문

目的：将本室构建与制备的双功能分子白介素-2-粒细胞-巨噬细胞集落生长因子（IL2-GMCSF）蛋白作用于肿瘤条件培养基（TCM）环境中的树突状细胞系（DC），检测其对DC细胞的活化，探讨其用于活化DC、进行抗肿瘤免疫治疗的可能性。方法制备小鼠黑色素瘤B16F10细胞系的TCM，用以培养DC2.4细胞，同时分别添加IL2-GMCSF、GM-CSF、IL-2、或IL-2与GM-CSF组合使用。24 h后，检测DC2.4细胞的吞噬与增殖活性、细胞成熟表型、细胞因子分泌与信号通路活化。结果 DC2.4细胞具有未成熟DC的特征，在TCM培养条件下吞噬能力增强、但增殖活性显著受抑，TCM对DC成熟表型表面标志的表达有一定促进作用，并促进单核与DC来源的趋化因子（MDC），但抑制DC的IL-12分泌。与之相反，IL2-GMCSF主要借助其GM-CSF活性，促进DC2.4细胞的吞噬与增殖活性，并促进DC进一步成熟，且高表达IL-12与MDC。与GM-CSF相比，IL2-GMCSF可诱导更高的炎性NF-κB通路活化水平，而抑制调节性STAT3通路的活化。结论与GM-CSF单独作用相比，IL2-GMCSF可更好地促进肿瘤免疫抑制环境中的DC活化，有望成为有效的临床抗肿瘤治疗手段。
목적：장본실구건여제비적쌍공능분자백개소-2-립세포-거서세포집락생장인자（IL2-GMCSF）단백작용우종류조건배양기（TCM）배경중적수돌상세포계（DC），검측기대DC세포적활화，탐토기용우활화DC、진행항종류면역치료적가능성。방법제비소서흑색소류B16F10세포계적TCM，용이배양DC2.4세포，동시분별첨가IL2-GMCSF、GM-CSF、IL-2、혹IL-2여GM-CSF조합사용。24 h후，검측DC2.4세포적탄서여증식활성、세포성숙표형、세포인자분비여신호통로활화。결과 DC2.4세포구유미성숙DC적특정，재TCM배양조건하탄서능력증강、단증식활성현저수억，TCM대DC성숙표형표면표지적표체유일정촉진작용，병촉진단핵여DC래원적추화인자（MDC），단억제DC적IL-12분비。여지상반，IL2-GMCSF주요차조기GM-CSF활성，촉진DC2.4세포적탄서여증식활성，병촉진DC진일보성숙，차고표체IL-12여MDC。여GM-CSF상비，IL2-GMCSF가유도경고적염성NF-κB통로활화수평，이억제조절성STAT3통로적활화。결론여GM-CSF단독작용상비，IL2-GMCSF가경호지촉진종류면역억제배경중적DC활화，유망성위유효적림상항종류치료수단。
Objective To test the effect of bifunctional molecule IL2-GMCSF in promoting the activation of dendritic cells (DCs) cultured in tumor conditioned medium. Methods We prepared a tumor conditioned medium using mouse melanoma cell line B16F10 supplemented with IL2-GMCSF, GM-CSF, IL-2, or the combination of the latter two. After culturing mouse DC cell line DC2.4 in the conditioned medium for 24 h, the DCs were examined for phagocytosis, proliferation, maturation phenotype, cytokine secretion, and signal pathway activation. Results DC2.4 cells displayed characteristics of immature DCs. After cell culture in the conditioned medium, the cells showed enhanced phagocytosis but significantly suppressed cell proliferation activity. Culture in the conditioned medium also promoted DC cell maturation and secretion of macrophage-derived chemokine (MDC), but inhibited IL-12 secretion. Supplementation of the conditioned medium with IL2-GMCSF promoted phagocytosis, proliferation, maturation, and cytokine (including both IL-12 and MDC) secretion of DC2.4 cells. Compared with GM-CSF, IL2-GMCSF induced a higher level of NF-κB signal pathway activation but suppressed STAT3 activation. Conclusion Compared with GM-CSF, IL2-GMCSF can better promote DC activation in the context of tumor-induced immune suppression, and thus shows potentials in anti-tumor therapy.