【作者】 刘春永；

【导师】 何贤辉；

【作者基本信息】 暨南大学 ， 免疫学， 2010， 硕士

【摘要】 目的：探讨尿酸钠作为佐剂对BALB/c小鼠体液免疫和细胞免疫应答的影响及机制。方法：1.利用尿酸钠悬浮液为佐剂、天花粉蛋白(TCS)为免疫原对BALB/c小鼠进行免疫,以酶联免疫测定法检测特异性抗体IgG的效价。2.体外诱导小鼠树突状细胞(DC),流式细胞术分析DC表型,评价尿酸钠体外对DC成熟的效应。3.以二硝基氟苯建立迟发型超敏反应(DTH)模型,分析尿酸钠在体内对细胞免疫应答的影响。结果：1.传统弗氏佐剂可极大地增强TCS对小鼠的血清IgG效价,而尿酸钠对IgG效价不但没有促进,反而有明显的抑制作用。结果提示,尿酸钠佐剂对抗体应答不但没有促进,与单独使用免疫原相比,抗体应答反而明显降低。2.从小鼠骨髓中分离得到的骨髓细胞,经rmGM-CSF和rmIL-4因子诱导,光学显微镜和原子力显微镜观察到明显的DC特征,提示DC诱导成功。流式细胞术分析结果显示,尿酸钠对DC表达CD11c和CD83没有影响,但可明显提高MHCⅡ的表达水平,表明尿酸钠对DC的成熟具有明显的促进作用。3.采用0.5％DNFB腹部致敏,0.2％DNFB耳廓激发的方法成功建立小鼠DTH模型。尿酸钠悬浮液对DTH模型小鼠耳组织肿胀、胸腺指数和脾脏指数有促进和提高作用；光学显微镜下观察小鼠耳廓切片,显示淋巴细胞浸润程度显著增加；MTS法检测结果显示,尿酸钠对ConA刺激的淋巴结和脾脏淋巴细胞有显著增殖作用。尿酸钠悬浮液能明显增强DNFB诱导的DTH炎症反应。结论：尿酸钠悬浮液作为佐剂,对细胞免疫有显著的增强作用,而对体液免疫应答却有一定的抑制作用,提示该佐剂在增强疫苗的细胞免疫应答中有潜在应用前景。更多还原

【Abstract】 AIM:To investigate the effects of uric acid sodium salt (UANa) as an adjuvant on humoral and cellular immune responses of the BALB/c mice.METHODS:1. BALB/c mice were immunized with trichosanthin (TCS) as an antigen together with UANa suspension as an adjuvant. The antibody titers of IgG were detected by enzyme-linked immunosorbent assay.2. Dendritic cells (DC) were induced in vitro, the phenotypes of DC were analyzed by flow cytometry and the effect of UANa on DC maturity was evaluated.3. Delayed-type hypersensitivity (DTH) model was used to analyze the effect of UANa on cellular immune responses in vivo. The in vitro proliferation was determined by ConA stimulation.RESULTS:1. Freund’s adjuvant could significantly enhance the serum IgG titer in mice against TCS, while UANa adjuvant failed to promote IgG titer but greatly restrain it. The results suggest that UANa adjuvant failed to promote the antibody response but significantly reduced antibody response when compared with TCS only.2. Bone marrow cells were isolated from mice’s bone marrow and stimulated by rmGM-CSF and rmIL-4, DC characteristics were distincted by Optical Microscopy and Atomic Force Microscopy, suggesting that DC was induced successfully.Flow cytometry analysis showed that UANa has no effect on CD11c and CD83 expression on DC, but could significantly enhance the expression level of MHCⅡmolecules, indicating that UANa has obviously promoted DC maturation.3. Abdomen with 0.5% DNFB sensitization, mouse DTH model was successfully established. ear tissue swelling and the thymus index and spleen index were promoted or enhanced by UANa suspension on DTH model; Biopsy of mouse ear were observed under optical microscope, implying that lymphocyte infiltration increased significantly; The lymphocytes proliferation capacity of lymph nodes and spleen from DTH mice was assayed by MTS colorimetry, the rusults showed that UANa Suspension could enhanced it notablely. In conclusion, UANa Suspension significantly increased DNFB-induced DTH inflammatory response in vitro.CONCLUSIONS:UANa suspension adjuvant could significantly enhance the cellular immune response but inhibit the humoral immune response to a certain degree, suggesting that UANa has potential application in enhancing the cellular immunity of a vaccine.更多还原