Ex Vivo Lung Function Measurements in Precision-Cut Lung Slices (PCLS) from Chemical Allergen-Sensitized Mice Represent a Suitable Alternative to In Vivo Studies
Ex Vivo Lung Function Measurements in Precision-Cut Lung Slices (PCLS) from Chemical Allergen-Sensitized Mice Represent a Suitable Alternative to In Vivo Studies
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DOI：
10.1093/toxsci/kfn178
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发表时间：
2008-12-01
影响因子：
3.8
通讯作者：
Braun, A.
中科院分区：
文献类型：
--
作者：
Henjakovic, M.;Martin, C.;Braun, A.
许多工业化学品都能引起呼吸道过敏反应。因此，迫切需要一种方法来识别和表征化学物质在肺中的生物作用。在这里，我们提出了一种简单，可靠的替代方法来测量暴露于化学过敏原后的体外肺功能变化，并将其与工业化学品trimellitic anhy酐（TMA）和2,4-二硝基氯苯（DNCB）致敏后的侵入性体内测量进行比较。雌性BALB/c小鼠被呼吸道过敏原TMA和接触过敏原DNCB表皮致敏。对TMA和DNCB的早期过敏反应在体内和离体实验中记录于吸入干燥的标准化气溶胶或暴露于溶解的过敏原后的第21天。第二天在体内和离体测量气道对增加剂量的甲胆碱（MCh）的高反应性（AHR）。支气管肺泡灌洗（BAL）用于局部炎症的免疫学表征。tma致敏小鼠在体内（ED50: 0.06 μ g MCh，对照组为0.21 μ g MCh）和PCLS中（EC50: 0.24 μ M MCh，对照组为0.4 μ M MCh）对MCh表现出AHR。tma处理的动物在过敏原刺激24小时后，BAL液中嗜酸性粒细胞数量增加(12.8.10(4)比0.7.10(4))，eotaxin-2浓度升高（994 pg/ml比167 pg/ml）。相比之下，用DNCB致敏后，这些参数都没有变化。本研究提示，TMA、DNCB等低分子量过敏原对PCLS体外肺功能的影响反映了体内情况。
A wide range of industrial chemicals can induce respiratory allergic reactions. Hence, there is an urgent need for methods identifying and characterizing the biological action of chemicals in the lung. Here, we present an easy, reliable alternative method to measure lung function changes ex vivo after exposure to chemical allergens and compare this to invasive in vivo measurements after sensitization with the industrial chemicals trimellitic anhydride (TMA) and 2,4-dinitrochlorobenzene (DNCB). Female BALB/c mice were sensitized epicutaneously with the respiratory allergen TMA and the contact sensitizer DNCB. The early allergic response to TMA and DNCB was registered in vivo and ex vivo on day 21 after inhalational challenge with dry standardized aerosols or after exposure of precision-cut lung slices (PCLS) to dissolved allergen. Airway hyperresponsiveness (AHR) to increasing doses of methacholine (MCh) was measured on the next day in vivo and ex vivo. Bronchoalveolar lavage (BAL) was performed for immunological characterization of local inflammation. TMA-sensitized mice showed AHR to MCh in vivo (ED50: 0.06 mu g MCh vs. 0.21 mu g MCh in controls) and in PCLS (EC50: 0.24 mu M MCh vs. 0.4 mu M MCh). TMA-treated animals showed increased numbers of eosinophils (12.8.10(4) vs. 0.7.10(4)) and elevated eotaxin-2 concentrations (994 pg/ml vs. 167 pg/ml) in BAL fluid 24 h after allergen challenge. In contrast, none of these parameters differed after sensitization with DNCB. The present study suggests that the effects of low molecular weight allergens, like TMA and DNCB, on ex vivo lung functions tested in PCLS reflect the in vivo situation.