胞嘧啶-磷酸-鸟嘌呤（CpG）寡核苷酸在非哮喘性慢性鼻-鼻窦炎中的免疫调节作用：一种外植体模型

Immunomodulatory effect of cytosine-phosphate-guanosine (CpG)-oligonucleotides in nonasthmatic chronic rhinosinusitis: an explant model.

作者信息

Tan Lorwai, Rogers Trisha J, Hatzirodos Nick, Baker Leonie M, Ooi Enghooi, Wormald Peter-John

机构信息

Department of Surgery-Otorhinolaryngology Head and Neck Surgery, University of Adelaide, The Queen Elizabeth Hospital, Adelaide, Australia.

出版信息

Am J Rhinol Allergy. 2009 Mar-Apr;23(2):123-9. doi: 10.2500/ajra.2009.23.3279.

DOI:10.2500/ajra.2009.23.3279

PMID:19401034

Abstract

BACKGROUND

The use of cytosine-phosphate-guanosine-oligodeoxynucleotides (CpG-ODNs) or immunostimulatory sequences (ISSs) in the treatment of airway diseases is gaining interest. Binding of the CpG-ODN ligand to Toll-like receptor 9 (TLR9) triggers a shift from a Th2- to a Th1-type response in the target tissue. In this study, we explored the potential use of CpG-ODN to dampen the predominantly Th2-driven chronic inflammatory state in our cohort of patients.

METHODS

An in vitro explant model comprising of sinonasal tissue from patients with asthma (n = 12) and without asthma (n = 11) were stimulated with CpG-ODN or Staphylococcus aureus enterotoxin B (SEB) or CpG-ODN in combination with SEB for 48 hours. Ten of the 12 asthma patients had nasal polyps. RNA was extracted for multiplex real-time reverse transcription polymerase chain reaction analysis and the 2(-delta deltaC(T)) method used to determine interleukin (IL)-5, p35 IL-12, interferon (IFN) gamma, and TLR9 expression levels.

RESULTS

CpG-ODN significantly reduced IL-5 mRNA expression in patients without asthma (p = 0.0379) but not in the asthma-associated group. SEB alone caused an increase in IL-5 levels that could be dampened when CpG-ODN was added in combination with SEB. Significant differences in mean IL-5 expression levels between the asthmatic and nonasthmatic categories were detected (Welch t-test; **p = 0.0041). Asthmatic and nonasthmatic patients present as two distinct categories as reflected by significant differences in their IL-5 response to CpG-ODN (F = 11.93; ***p = 0.0008), SEB (F = 41.34; *p = 0.0476) and CpG-ODN with SEB (F = 13.2; *p = 0.0114). In contrast, no significant differences were observed in the expression levels of IL-12, IFN-gamma, and TLR9.

CONCLUSION

Localized application of CpG-ODN on its own or in combination with SEB may potentially reduce the expression of the proinflammatory cytokine IL-5 in nonasthmatic patients and may be further developed as an immunotherapeutic agent.

摘要

背景

胞嘧啶 - 磷酸 - 鸟嘌呤 - 寡脱氧核苷酸（CpG - ODNs）或免疫刺激序列（ISSs）在气道疾病治疗中的应用正受到关注。CpG - ODN配体与Toll样受体9（TLR9）结合会引发靶组织中从Th2型反应向Th1型反应的转变。在本研究中，我们探讨了CpG - ODN在我们的患者队列中抑制主要由Th2驱动的慢性炎症状态的潜在用途。

方法

用CpG - ODN或金黄色葡萄球菌肠毒素B（SEB）或CpG - ODN与SEB联合刺激由哮喘患者（n = 12）和非哮喘患者（n = 11）的鼻窦组织组成的体外外植体模型48小时。12名哮喘患者中有10名患有鼻息肉。提取RNA用于多重实时逆转录聚合酶链反应分析，并使用2（-ΔΔC（T））方法测定白细胞介素（IL）-5、p35 IL - 12、干扰素（IFN）γ和TLR9的表达水平。

结果

CpG - ODN显著降低了非哮喘患者中IL - 5 mRNA的表达（p = 0.0379），但在哮喘相关组中未降低。单独使用SEB会导致IL - 5水平升高，当与CpG - ODN联合添加时这种升高可被抑制。检测到哮喘组和非哮喘组之间平均IL - 5表达水平存在显著差异（Welch t检验；**p = 0.0041）。哮喘患者和非哮喘患者表现为两个不同的类别，这通过他们对CpG - ODN（F = 11.93；***p = 0.0008）、SEB（F = 41.34；*p = 0.0476）以及CpG - ODN与SEB联合（F = 13.2；*p = 0.0114）的IL - 5反应的显著差异得以体现。相比之下，IL - 12、IFN - γ和TLR9的表达水平未观察到显著差异。