补肾回阳方体内前期免疫激活对DC-CIK细胞扩增和杀伤效应的影响

The DC-CIK cells which are the first choice of the anti-tumor adoptive immunotherapy can be amplificated hundreds of times in vitro. However, the clinical effect is still not good enough. It is all because that we all focus on the amplification in vitro but ignoring the inside activation. The ancient Huangdineijing says that the five visceral organs are interpromoting related and the Weiqi is originated from the kidney. Due to the theory we introduce the hypothesis that the essence of the immunity is not in the immunity itself but outside the immunity. To enhance the immunity, we need to pay more attention to the early activation inside the body. Our early study shows that the Warming yang medicine could increase the cellular immunity; and someone also found the target of these medicines was hypothalamus. These findings can verify this hypothesis. Although the Bushen Huiyang Decociton has anti-cancer effect, the mechanism still needs to be verified whether it enhanced the later tumoricidal effect of DC-CIK by increase the proficiency of immunity precursor cells through earlier activation inside the body. The project will imitate tumor patients in immunosuppressed state to establish a mouse model and study traditional Chinese medicine on the immune cells in the immune function subsets, cytokines and substance P in the pre-activated in vivo. At the same time, we will separate the mouse lymphocytes of the late cytokine re-activated DC-CIK cells in vitro amplification and study the Chinese medicine pre-excited DC-CIK anti-leukemia synergies in cell level and animal level.We intend to explain the essence of immune regulation outside the immunity itself. Therefore we can find a news break in the discovery of immunity activation.

抗肿瘤过继免疫治疗首选的DC-CIK细胞虽能体外成百倍扩增但疗效不佳，这与当前偏重于体外扩增却忽视体内免疫激活有关。据《黄帝内经》五脏相生、卫气根于肾的理论，我们提出免疫之根不在免疫本身而在免疫之外的假设，增强免疫应先重视前期的体内激活。先前发现温阳中药可增强细胞免疫，另有研究发现其作用于下丘脑，初步证实了此假设。补肾回阳方等具有临床抗癌良效，但其是否能先通过体内前期激发提高免疫前体细胞潜能，然后增强后期的DC-CIK体外扩增而发挥杀癌效应等需要探明。本项目将模仿肿瘤病人免疫抑制状态建立小鼠模型，研究中药预先体内激活该小鼠免疫功能后对其免疫细胞亚群、细胞因子和Ｐ物质的影响。同时，分离该小鼠淋巴细胞进行后期细胞因子再激活的DC-CIK细胞体外扩增，在细胞水平和动物整体水平研究中药前期激发对DC-CIK抗白血病的增效作用 ，阐明从免疫之外调免疫的科学本质，以求在免疫激活新方法学研究上有所突破。

抗肿瘤过继免疫治疗首选的DC-CIK细胞虽能体外成百倍扩增但疗效不佳，这与当前偏重于体外扩增却忽视体内免疫激活有关。据《黄帝内经》五脏相生、卫气根于肾的理论，我们提出免疫之根不在免疫本身而在免疫之外的假设，增强免疫应先重视前期的体内激活。先前发现温阳中药可增强细胞免疫，另有研究发现其作用于下丘脑，初步证实了此假设。补肾回阳方等具有临床抗癌良效，但其是否能先通过体内前期激发提高免疫前体细胞潜能，然后增强后期的DC-CIK体外扩增而发挥杀癌效应等需要探明。本项目模仿肿瘤病人免疫抑制状态建立小鼠免疫抑制模型，可见小鼠毛发光泽较暗淡，脱落明显，呈稀疏样，可见模型组小鼠有明显的湿尾现象，大便呈稀溏状，血清T3水平明显降低，模型组中性粒细胞降低。模型对照组外周血 IL-2、IL-6、IFN-γ显著降低， TNF 降低；模型组外周血 IL-4、IL-10、IL-17 均显著性升高。表明肾阳虚模型小鼠免疫功能低于空白对照组，即造模成功。三阳血傣组和补肾回阳组的中性粒细胞均显著性降低。三阳血傣组和补肾回阳组淋巴细胞均显著性升高。说明补肾回阳方能够增强肾阳虚模型小鼠的免疫功能。三阳血傣组、补肾回阳组的 CD4+、CD4/CD8+数值均明显升高，补肾回阳组的 CD8+细胞升高。说明补肾回阳方通过对 T细胞平衡的调节作用对肾阳虚小鼠发挥保护作用。三阳血傣组、补肾回阳组外周血 IL-2 、TNF 均显著性升高。补肾回阳方低剂量 组、补肾回阳方高剂量组外周血 IL-6 显著性升高。三阳血傣组、补肾组外周血 IFN-γ 与模型对照组相比显著性升高。三阳血傣组、补肾回阳组外周血 IL-17 均显著性升高。同时，分离该小鼠淋巴细胞进行后期细胞因子再激活的DC-CIK细胞体外扩增，补肾回阳组、三阳血傣组CIK细胞增殖能力明显提高，尤其是补肾回阳组细胞杀伤活性明显提高。说明补肾回阳组前期激发对DC-CIK抗白血病的增效作用 ，部分阐明了从免疫之外调免疫的科学本质。