通过重组腺相关病毒载体介导的反义白介素4基因转染对过敏症大鼠气道重构的影响
2010/11/04
摘要
背景：包括白介素4（IL-4）在内的Th2细胞因子被认为在哮喘气道重构中起到重要作用。
目的：我们前期研究显示，携带反义IL-4基因的重组腺相关病毒载体（rAAV-asIL4）能显著抑制过敏性哮喘大鼠气道IL-4的表达及气道炎症。本试验通过rAAV-asIL4 载体抑制IL-4表达，研究抗IL-4治疗在预防过敏性哮喘气道重构中的作用。
方法：通过尾静脉注射卵清蛋白（OVA），建立大鼠过敏性哮喘模型，并转染rAAV-asIL4载体。采用酶联免疫分析法分析支气管肺泡灌洗液（BALF）中IL-4含量。计数BALF中嗜酸性粒细胞含量。通过免疫组织化学法，研究支气管周围转化生长因子b1（TGF-b1）和TGF-b2阳性细胞，天狼星红染色检测基底膜下胶原沉积。收集肺组织进行组织学分析，检测总支气管面积（WAt）和气道平滑肌面积（WAm）。
结果：rAAV-asIL4转染能显著降低哮喘大鼠BALF中IL-4含量。BALF中嗜酸性粒细胞、支气管周围TGF-b1和TGF-b2阳性细胞也减少。此外，rAAV-asIL4处理能抑制气道天狼星红染色面积，增加WAt和WAm。
结论：这些结果显示，rAAV-asIL4转染能抑制与气道炎症相关的气道重构过程。本研究可能为rAAV-asIL4作为治疗哮喘相关气道重构提供证据。
（陈欣 审校）
J Asthma. 2010 Sep 13. [Epub ahead of print]
The effects of antisense interleukin-4 gene transferred by recombinant adeno-associated virus vector on the airway remodeling in allergic rats.
Cao Y, Zeng D, Song Q, Cao C, Xie M, Liu X, Xiong S, Xu Y, Xiong W.
Department of Respiratory Medicine, Tongji Hospital, Key Laboratory of Pulmonary Diseases of the Ministry of Health of China, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Abstract
Background: Th2-derived cytokines, including interleukin-4 (IL-4), are considered to play an important role in the development of airway remodeling of asthma.
Objectives: Our previous study has demonstrated that a recombinant adeno-associated virus containing antisense against IL-4 gene (rAAV-asIL4) vector could significantly suppress the expression of IL-4 protein and airway inflammation in the rat models of allergic asthma. In this study, we applied the rAAV-asIL4 vector to allergic rats to investigate the effects of anti-IL4 therapy on airway remodeling in allergic asthma.
Methods: rAAV-asIL4 was used to infect the ovalbumin (OVA)-sensitized and challenged rats by tail-vein injection. IL-4 protein in bronchoalveolar lavage fluid (BALF) was detected by enzyme-linked immunosorbent assay. The number of eosinophils in BALF was counted. Transforming growth factor-beta1 (TGF-beta1) and TGF-beta2-positive cells in the peribronchial space were detected by immunohistochemical staining, and collagen deposition beneath the basement membrane was detected by Sirius red stain. The lung tissues were collected for histologic analysis of total bronchial wall area (W(At)) and airway smooth muscle area (W(Am)).
Results: rAAV-asIL4 significantly decreased IL-4 protein in BALF of OVA-sensitized and challenged rats. The number of eosinophils in BALF, the TGF-beta1 and TGF-beta2-positive cells in the peribronchial space were also suppressed. Moreover, the rAAV-asIL4 treatment inhibited the area of Sirius red staining in airways and the increase in W(At) and W(Am).
Conclusion: These results suggest that rAAV-asIL4 may attenuate the airway remodeling process relevant to the inhibition of airway inflammation. This study provides elementary evidence for the potential utility of rAAV-asIL4 as an approach to gene therapy for asthmatic airway remodeling.
J Asthma. 2010 Sep 13. [Epub ahead of print]
背景：包括白介素4（IL-4）在内的Th2细胞因子被认为在哮喘气道重构中起到重要作用。
目的：我们前期研究显示，携带反义IL-4基因的重组腺相关病毒载体（rAAV-asIL4）能显著抑制过敏性哮喘大鼠气道IL-4的表达及气道炎症。本试验通过rAAV-asIL4 载体抑制IL-4表达，研究抗IL-4治疗在预防过敏性哮喘气道重构中的作用。
方法：通过尾静脉注射卵清蛋白（OVA），建立大鼠过敏性哮喘模型，并转染rAAV-asIL4载体。采用酶联免疫分析法分析支气管肺泡灌洗液（BALF）中IL-4含量。计数BALF中嗜酸性粒细胞含量。通过免疫组织化学法，研究支气管周围转化生长因子b1（TGF-b1）和TGF-b2阳性细胞，天狼星红染色检测基底膜下胶原沉积。收集肺组织进行组织学分析，检测总支气管面积（WAt）和气道平滑肌面积（WAm）。
结果：rAAV-asIL4转染能显著降低哮喘大鼠BALF中IL-4含量。BALF中嗜酸性粒细胞、支气管周围TGF-b1和TGF-b2阳性细胞也减少。此外，rAAV-asIL4处理能抑制气道天狼星红染色面积，增加WAt和WAm。
结论：这些结果显示，rAAV-asIL4转染能抑制与气道炎症相关的气道重构过程。本研究可能为rAAV-asIL4作为治疗哮喘相关气道重构提供证据。
（陈欣 审校）
J Asthma. 2010 Sep 13. [Epub ahead of print]
The effects of antisense interleukin-4 gene transferred by recombinant adeno-associated virus vector on the airway remodeling in allergic rats.
Cao Y, Zeng D, Song Q, Cao C, Xie M, Liu X, Xiong S, Xu Y, Xiong W.
Department of Respiratory Medicine, Tongji Hospital, Key Laboratory of Pulmonary Diseases of the Ministry of Health of China, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Abstract
Background: Th2-derived cytokines, including interleukin-4 (IL-4), are considered to play an important role in the development of airway remodeling of asthma.
Objectives: Our previous study has demonstrated that a recombinant adeno-associated virus containing antisense against IL-4 gene (rAAV-asIL4) vector could significantly suppress the expression of IL-4 protein and airway inflammation in the rat models of allergic asthma. In this study, we applied the rAAV-asIL4 vector to allergic rats to investigate the effects of anti-IL4 therapy on airway remodeling in allergic asthma.
Methods: rAAV-asIL4 was used to infect the ovalbumin (OVA)-sensitized and challenged rats by tail-vein injection. IL-4 protein in bronchoalveolar lavage fluid (BALF) was detected by enzyme-linked immunosorbent assay. The number of eosinophils in BALF was counted. Transforming growth factor-beta1 (TGF-beta1) and TGF-beta2-positive cells in the peribronchial space were detected by immunohistochemical staining, and collagen deposition beneath the basement membrane was detected by Sirius red stain. The lung tissues were collected for histologic analysis of total bronchial wall area (W(At)) and airway smooth muscle area (W(Am)).
Results: rAAV-asIL4 significantly decreased IL-4 protein in BALF of OVA-sensitized and challenged rats. The number of eosinophils in BALF, the TGF-beta1 and TGF-beta2-positive cells in the peribronchial space were also suppressed. Moreover, the rAAV-asIL4 treatment inhibited the area of Sirius red staining in airways and the increase in W(At) and W(Am).
Conclusion: These results suggest that rAAV-asIL4 may attenuate the airway remodeling process relevant to the inhibition of airway inflammation. This study provides elementary evidence for the potential utility of rAAV-asIL4 as an approach to gene therapy for asthmatic airway remodeling.
J Asthma. 2010 Sep 13. [Epub ahead of print]
