摘要
目的 探讨Toll样受体9（toll like receptor 9,TLR9）配体CpG寡脱氧核苷酸（CpG oligodeoxynucleotides）鼻内预免疫对豚鼠变应性鼻炎（allergic rhinitis,AR）的抑制作用.方法 50只实验豚鼠采用随机数宁表法分为5组,每组10只,分别为：空白对照组（简称对照组）、AR模型组（AR组）、CpG组、CpG＋卵清蛋白组（ovalbumin,OVA,CpG＋OVA组）、CpG模拟序列＋OVA组（CpG-M＋OVA组）.AR组及对照组给予生理盐水预免疫;CpG组给予5.2 μg CpG826;CpG＋OVA组给予5.2μg CpG＋4.8μg OVA;CpG-M＋OVA组给予不具免疫活性的CpG1982加OVA滴鼻.4个实验组动物依次进行腹腔OVA和氢氧化铝凝胶基础致敏,2次鼻腔激发,对照组则给予生理盐水.观察鼻黏膜组织损伤情况及嗜酸粒细胞计数.酶联免疫吸附法（ELISA）检测血清及鼻黏膜组织匀浆中白细胞介素（IL）5及γ干扰素（interferon γ,IFN-γ）的水平.免疫荧光（immunofluorescence,IF）观察细胞间黏附分子（intercellular adhesion molecule,ICAM）1在鼻黏膜上皮组织的表达.采用SPSS 11.0软件对数据进行分析.结果 CpG组动物的喷嚏,抓鼻症状要轻于AR组（Z值分别为3.412、3.523,P值均〈0.05）,CpG＋OVA组症状轻于CpG-M＋OVA组（Z值分别为3.984、4.452,P值均〈0.05）.CpG组豚鼠鼻黏膜组织嗜酸粒细胞平均（（x）±s,下同）计数为（20.0±9.6）个,低于模型组的（53.5±19.8）个,CpG＋OVA组豚鼠鼻黏膜组织嗜酸粒细胞计数为（9.5±5.7）个,低于CpG-M＋OVA组的（49.2±18.9）个,差异有统计学意义（q值分别为3.785、4.576,P值均〈0.05）.ELISA结果显爪CpG组血清及鼻黏膜组织匀浆中IL-5水平低于AR组（q值分别为3.890、4.019,P值均〈0.05）,而CpG＋OVA组低于CpG-M＋OVA组（q值分别为4.128、4.245,P值均〈0.05）,而各组IFN-γ的水平差异尢统计学意义（F值分别为2.078、2.102,P值均〉0.05）.免疫荧光结果赤示CpG组鼻黏膜组织ICAM-1的表达水平要低于AR组,CpG＋OVA组低于CpG-M＋OVA组（Z值分别为3.697、3.765,P值均〈0.05）.结论 CpG寡脱氧核苷酸鼻内预免疫是一种行之有效的抑制鼻变态反应发生的方法.
Objective To evaluate the therapeutic effect of intranasal oligodeoxynucleotides with CpG motifs （CpG ODN） in prevention of allergic rhinitis in juvenile guinea pigs. Methods Juvenile guinea pigs aged from 7 to 10 weeks were administrated with CpG ODN alone or combined with OVA at single dose concentration intranasally （on day 0, 5, 10, 15 in sequence） while control and blank group were administrated with saline. Both experimental and control animals were again sensitized by OVA （on day 18, 25）, and 14 days after second sensitization animals were challenged by OVA intranasally （on days 39 and 46）. Two hours after challenge, the animals were sacrificed. Then Hemotoxin and Eosin stain were carried out to analyze local eosinophilic reactions and nasal lesions. Local and systemic cytokines interleukin IL-5 and IFN-γ levels were examined by ELISA. Immunoflourescence was carried out with ICAM-1 antibody. Statistical analysis was performed using a SPSS 11.0 software. Results In CpG ODN-administration or CpG ODN with OVA-administration group allergic rhinitis symptoms were not as severe as model control group （P〈0.05）. Compared with the model control group, CpG ODN-administration did not increase production of OVA-specific Thl cytokine IFN-γ but decreased productions of ovalbumin-specific Th2 cytokines IL-5 both in serum and nasal specimen（q value were 3.890 and 4.019, P〈0. 05）. Moreover, nasal lesions with infiltration of mean （x ±s） eosinophils（20. 0 ±9. 6） in CpG group animal were prominently reduced by the CpG ODN-treatment compared with the control animals （53. 5 ± 19. 8） and CpG ＋ OVA group （9. 5 ±5. 7） were lower than CpG-M ＋ OVA group （49. 2 ± 18. 9）, the differences were significant （q value were 3.785 and 4. 576, P 〈0. 05）. Immunoflourescence results showed lower ICAM-1 expression in nasal specimen of CpG group compared with model group and CpG plus OVA group animal to CpG mimics plus OVA group （Z value were 3.697 and 3.765, P 〈 0.05 ）. Conclusions Intranasal administration of CpG oligodeoxynucleotides with or without allergen may be an effective way to prevent the development of allergic rhinitis.
出处
《中华耳鼻咽喉头颈外科杂志》
CAS
CSCD
北大核心
2010年第6期471-476,共6页
Chinese Journal of Otorhinolaryngology Head and Neck Surgery
基金
国家自然科学基金面上项目（30772409）
卫生部临床学科重点资助项目（07090138）
吉林省科技发展项目（200705275）
