以纳米粒子为佐剂的屋尘螨疫苗免疫治疗的实验研究

发布时间：2018-06-02 02:02

本文选题：哮喘 + 纳米粒子　；参考：《南昌大学》2006年硕士论文

【摘要】：目的：建立屋尘螨哮喘动物模型，比较分别用屋尘螨粗浸液、重组Der p2、包被rDer p2纳米粒子进行免疫治疗的疗效，为变态反应性疾病的免疫治疗提供新的思路。方法：首先利用PLGA(聚乳酸-羟乙酸)制备包被重组Der p2的纳米粒子。其次，进行实验动物分组：近交系清洁级6～8周龄BALB／c小鼠40只，体重18～20克，随机分配为：A阴性对照组，B阳性对照组，C粗浸液治疗组(简称：粗浸液组)，D rDer p2治疗组(简称：rDer p2组)，E包裹rDer p2纳米粒子治疗组(简称：纳米粒子组)，每组8只，雌雄各半。哮喘模型建立及干预措施：第0、7、14天腹膜内注射(腹腔注射，i．p．)0.1mg／ml屋尘螨变应原粗浸液与10mg／ml Al(OH)_3生理盐水的混悬液100μl：第28～30天对C、D、E组分别给予i．p．1mg／ml屋尘螨粗浸液100μl、1mg／ml rDer p2溶液100μl、1mg／ml包被rDer p2纳米粒子溶液100 μl进行治疗；第44～49天B、C、D、E组给予5mg／ml屋尘螨粗浸液100μl鼻腔滴入(i．n．)方式激发，A组一直相应给予生理盐水处理；第51天处死各组动物。以酶联免疫吸附试验(ELISA)的方法检测各组实验动物血清中IgE、IgGl、IgG2a的水平，以及支气管肺泡灌洗液(BALF)与脾细胞培养后上清液(SCCS)中细胞因子IL-4、IFN-γ及IL-10水平，并计数BALF中细胞总数与分类细胞数，制备肺组织病理切片以观察其病理改变，并选择视野用显微摄像仪拍照。结果：添加铝佐剂的屋尘螨变应原致敏的小鼠在特异性变应原的连续激发下，肺组织产生明显的炎症反应，表现为BALF中细胞总数增加以及炎症细胞所占比例及绝对计数升高，且以分泌Th2型细胞因子如IL-4为主。血清IgE、IgGl水平升高。肺组织病理切片观察到支气管、细支气管上皮下和小血管周围有明显的炎性细胞浸润，以嗜酸性粒细胞为主，，气道上皮结构紊乱，有部分脱落，部分可见粘液栓，上皮下充血水肿明显，肺泡间质增厚，也可见到
[Abstract]:Objective: to establish the animal model of house dust mite asthma and compare the efficacy of immunotherapy with the crude extract of house dust mite, recombinant Der p2 and rDer p2 nanoparticles, respectively, in order to provide a new idea for immunotherapy of allergic diseases. Methods: firstly, poly (lactic acid-glycolic acid) nanoparticles coated with recombinant Der p2 were prepared. Secondly, the experimental animals were divided into four groups: 40 inbred BALB/c mice of clean grade 6 and 8 weeks old, weighing 1820 grams. They were randomly divided into two groups: the control group with negative weight A and B positive control group with crude C crude extract group (abbreviated as the crude extract group) and the D rDer p2 treatment group (abbreviated as the rDer p2 group), which was coated with rDer p2 nanoparticles (8 rats in each group, half male and half female). Establishment of asthma model and intervention: intraperitoneal injection (i.p.lg / ml) intraperitoneal injection (i.p.) of 0.1 mg / ml crude extract of house dust mite allergen and 100 渭 l of 10mg/ml Al(OH)_3 normal saline solution on day 0, 714: 100 渭 lg / ml rDer p2 solution 100 渭 lg / ml, respectively, on day 2830, in group C + DX E RDer p2 nanoparticles were coated with 100 渭 l solution. On the 44th and 49th day, the rats in group E were given 100 渭 l crude extract of 5mg/ml house dust mites into their nasal cavity. Methods Group A was treated with normal saline and animals were killed on the 51st day. Elisa was used to detect the level of IgG2a in serum of experimental animals, and the levels of cytokines IL-4, IFN- 纬 and IL-10 in the supernatant of bronchoalveolar lavage fluid (BALF) and splenocytes. The total number of cells and the number of classified cells in BALF were counted, the pathological sections of lung tissue were prepared to observe the pathological changes, and the field of vision was photographed with a microscopic camera. Results: the mice sensitized with aluminum adjuvant caused by specific allergen produced obvious inflammatory reaction in lung tissue, which was manifested by the increase of the total number of cells, the proportion of inflammatory cells and the absolute count of inflammatory cells in BALF. And mainly secreted Th2 type cytokines such as IL-4. The level of IgGl in serum was increased. The inflammatory cell infiltration was observed in bronchus, bronchioles subcutaneously and around small vessels, mainly eosinophilic granulocytes. The airway epithelial structure was disordered, some of them fell off, and some mucus suppositories were observed. Epigastric hyperemia and edema, alveolar interstitial thickening, can also be seen
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R392

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