四种中药单体作为疫苗佐剂的体外筛选及原花青素作为肿瘤细胞疫苗佐剂在小鼠黑色素瘤模型中的应用

发布时间：2018-11-12 06:50

【摘要】：目的检测原花青素、天麻素、黄芩素和芹菜素对抗原提呈细胞RAW264.7和DC2.4的免疫刺激活性,并将免疫刺激活性最好的原花青素作为肿瘤细胞疫苗佐剂在小鼠皮下黑色素瘤模型中进行抗肿瘤研究,为肿瘤疫苗的研发提供一种新型可供选择的佐剂。方法体外实验以不同种类、不同浓度的中药单体分别刺激RAW264.7和DC2.4细胞,48 h后用抗体染色,流式细胞术检测两种抗原提呈细胞表面激活标志CD80,CD86,MHC T,MHC TT分子表达量变化；动物实验以紫外灭活的小鼠黑色素瘤细胞B16F10作为抗原,将肿瘤抗原与原花青素充分混合作为肿瘤细胞疫苗,采用肌肉注射的方式免疫荷瘤C57BL/6小鼠。结果体外实验结果显示不同种类、不同浓度的中药单体分别刺激RAW264.7和DC2.4细胞后,两种抗原提呈细胞表面CD80,CD86,MHC ⊥,MHC ⊥⊥分子不同程度表达上调,原花青素具有较强的免疫刺激活性；动物实验结果显示原花青素能够增强小鼠体内Dc的激活,原花青素作为肿瘤细胞疫苗佐剂免疫的荷瘤小鼠肿瘤生长速度减慢并且生存期延长,肿瘤细胞疫苗能够诱导小鼠产生高水平针对B16F10抗原的特异性的细胞杀伤,并且能够激活小鼠体内分泌高水平perforin、IFN-γ和TNF-α的CD8+T细胞；结果表明原花青素作为肿瘤细胞疫苗佐剂在小鼠黑色素瘤模型中能够诱导一定的抗肿瘤效果。结论原花青素是一种具有潜在应用价值的肿瘤细胞疫苗佐剂,本研究为原花青素作为疫苗佐剂的进一步应用提供了理论基础。
[Abstract]:Objective to detect the immunostimulatory activity of proanthocyanidins, gastrodin, baicalin and apigenin on antigen-presenting cells (RAW264.7 and DC2.4). Proanthocyanidins, which have the best immunostimulatory activity, were used as adjuvant of tumor cell vaccine in mouse subcutaneous melanoma model to provide a new adjuvant for the development of tumor vaccine. Methods RAW264.7 and DC2.4 cells were stimulated with different kinds and different concentrations of traditional Chinese medicine monomers in vitro. After 48 hours, antibody staining was used to detect the activation marker CD80,CD86,MHC T on the surface of the two antigen-presenting cells. The change of MHC TT expression; Mouse melanoma cell line B16F10 was used as antigen, tumor antigen and procyanidin were mixed as tumor cell vaccine, and C57BL/6 mice were immunized by intramuscular injection. Results after stimulation of RAW264.7 and DC2.4 cells with different kinds of Chinese medicine monomers and different concentrations of Chinese medicine monomer in vitro, the surface of the two antigen-presenting cells was up-regulated in different degrees, and the expression of CD80,CD86,MHC Karabakh and MHC was up-regulated in different degrees on the surface of the two antigen-presenting cells. Proanthocyanidins have strong immunostimulatory activity. The results of animal experiments showed that proanthocyanidins could enhance the activation of Dc in mice. The tumor growth rate and survival time of tumor-bearing mice immunized with proanthocyanidins as adjuvant of tumor cell vaccine were decreased and the survival time was prolonged. Tumor cell vaccine can induce high level of specific cytotoxicity against B16F10 antigen in mice and activate CD8 T cells secreting high levels of perforin,IFN- 纬 and TNF- 伪 in mice. The results showed that proanthocyanidins as adjuvant of tumor cell vaccine could induce anti-tumor effect in mouse melanoma model. Conclusion proanthocyanidin is a potential tumor cell vaccine adjuvant. This study provides a theoretical basis for the further application of proanthocyanidins as vaccine adjuvants.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R392

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