摘要
目的探讨螨变应原Der p2经聚乳酸-羟基乙酸共聚物(PLGA)包载合成的纳米疫苗特异性免疫治疗小鼠过敏性鼻炎的疗效及机制。方法取雄性SPF级BALB/c小鼠30只,4~6周龄。随机分为5组,即正常组、模型组、空白PLGA治疗组、螨抗原纳米疫苗治疗组(PLGA-DP2治疗组)及螨重组蛋白Der p2治疗组,每组6只。按常规方法建立尘螨过敏性鼻炎小鼠动物模型。建模成功后,空白PLGA治疗组、PLGA-DP2治疗组和螨重组蛋白Der p2治疗组分别使用空PLGA、PLGA-Der p2纳米疫苗(含50μg Der p2蛋白)和50μg螨重组蛋白Der p2经小鼠腹部皮下注射进行免疫治疗,每隔3 d注射1次,共治疗5次。治疗结束后,取螨变应原粗浸液200μg分别滴鼻激发小鼠,激发结束后处死小鼠。在小鼠激发后的30 min内,观察并记录各组小鼠鼻部症状(喷嚏次数、搔鼻及鼻分泌物量)评分情况;检测血清中过敏原特异性抗体(Ig E)及细胞因子[白细胞介素(IL)-2、IL-4、IL-10和γ干扰素(INF-γ)]水平;观察鼻黏膜组织形态学变化。结果螨抗原纳米疫苗PLGA-DP2载药量13.6%,包封率88.4%;纳米粒直径为200~400 nm。治疗后小鼠鼻部症状评分,PLGA-DP2治疗组鼻痒(0.67±0.52)分,喷嚏(1.00±0.63)分,清涕(1.67±0.41)分;螨重组蛋白Der p2治疗组分别为(0.83±0.41)分、(1.50±0.55)分、(0.83±0.75)分。明显低于模型组,且PLGA-DP2治疗组降低更明显(P<0.05)。PLGA-DP2治疗组和螨重组蛋白Der p2治疗组与模型组比较,血清特异性Ig E及IL-4水平方面均明显降低,PLGADP2治疗组降低更显著(P<0.05);而IL-2、IL-10和INF-γ水平显著增加,PLGA-DP2治疗组增加更显著(P<0.05)。PLGA-DP2治疗组小鼠鼻黏膜病理表现,上皮细胞基本排列整齐,组织结构清晰,相对模型组黏膜下层嗜酸性粒细胞、中性粒细胞等炎性细胞减少,鼻黏膜纤毛结构相对比较完整,黏膜基层肿大程度减轻;基本与正常组一致。结论尘螨变应原纳米疫苗加以PLGA为佐剂�
Objective To investigate the mechanism and efficacy of mite allergen Der p2-loaded poly lactic-co-glycolic acid（PLGA） for treatment of mice with allergic rhinitis. Method Thirty BALB/c mice of 4-6 weeks old were randomly divided into 5 groups（normal group, model group, blank PLGA treatment group, nanovaccne PLGA-DP2 treatment group and recombinant protein Der p2 treatment group, n = 6）. The mice model of dust mite allergic rhinitis was established by routine method. After successful modeling, blank PLGA treatment group received subcutaneous injection of blank PLGA, nanovaccine PLGA-DP2 treatment group received mite allergen profilin-loaded PLGA nanoparticles（containing 50 μg mite allergen）, and recombinant protein Der p2 treatment group received 50 μg recombinant protein Der p2, treated 5 times once every 3 days. After treatment,the mice were provocated with mite allergen crude infusion of 200 μg intranasally and sacrificed after provocation. Scores of nasal symptoms（numbers of sneezing, scratching nose and amount of nasal secretion） were recorded within 30 minutes after provocation. In addition, serum levels of allergen-specific antibodies（Ig E） and cytokines[interleukin（IL）-2, IL-4, IL-10 and interferon（INF-γ）] were measured, and morphological changes in nasal mucosa were observed. Results The drug loading of mite antigen nanovaccine PLGA-DP2 was 13.6 %,and encapsulation effeciency was 88.4 % with nanoparticles diameter of200-400 nm. After treatment, the score of nasal symptoms in nanovaccine PLGA-DP2 treatment group was nasal itching of0.67 ± 0.52, sneezing of 1.00 ± 0.63 and rhinocnesinus of 1.67 ± 0.41. The recombinant protein Der p2 treatment group was0.83 ± 0.41, 1.50 ± 0.55 and 0.83 ± 0.75, respectively. The scores were significantly lower than that of model group, and nanovaccine PLGA-DP2 treatment group decreased more significantly（P〈0.05）. Compared with model group, serum levels of Ig E and IL-4 reduced, and serum levels of IL-2, IL-10 and INF-γ increased in
出处
《生物医学工程与临床》
CAS
2017年第2期195-200,共6页
Biomedical Engineering and Clinical Medicine
基金
国家自然科学基金项目(31400856)
深圳市龙岗区科技发展资金项目(YLWS20140609111127924)
深圳市重点实验室基金项目(ZDSYS201506050935272)
关键词
纳米疫苗
PLGA
过敏性鼻炎
螨变应原
特异性免疫治疗
nanovaccine
poly lactic-co-glycolic acid（PLGA）
allergic rhinitis（AR）
mite allergen
special immunologic therapy（SIT）
