传统农场环境灰尘中的哮喘保护剂
2023/06/25
摘要
背景：在传统的欧洲和美国阿米什族奶牛场长大，与奶牛和干草密切接触，可以保护儿童免受哮喘的侵袭，而通过气道给予从这些奶牛场牛棚收集的灰尘中提取的提取物可以预防小鼠的过敏性哮喘。
目的：本研究试图开始确定农场衍生的哮喘保护剂。
方法：我们的工作沿着两条无偏倚、独立但互补的发现路径展开。通过比较核磁共振指纹图谱和差异蛋白质组学分析了保护性和非保护性农场（欧洲和阿米什牛棚与欧洲羊棚）的灰尘提取物。以生物活性为导向的粒度分级主要针对保护性阿米什牛棚粉尘提取物。在这两种途径中都使用了多种体外和体内功能测定。通过溶液中和凝胶中SDS-PAGE酶促消化/肽图谱，然后通过液相色谱/质谱对由此鉴定的一些蛋白质进行表征。这些蛋白质携带的物质通过非靶向液相色谱-高分辨率质谱法进行分析。
结果：从欧洲防护牛舍的灰尘提取物中富集了12种动植物来源的载体蛋白，包括牛脂肪运载蛋白Bos d2和气味结合蛋白。阿米什牛棚粉尘提取物中一种有效的哮喘保护成分（约占未分级提取物总碳含量的0.5%）含有七种动植物蛋白，包括Bos d2和气味结合蛋白，该蛋白富含植物、细菌和真菌的脂肪酸代谢产物。
结论：传统农场的动植物产生的蛋白质可以运输疏水性微生物和植物代谢产物。当输送到黏膜表面时，这些药物可能会调节气道反应。
Asthma-protective agents in dust from traditional farm environments
Mauricius Marques Dos Santos, Vadim Pivniouk, Bettina Rankl, Alesia Walker, Giulia Pagani, Norbert Hertkorn, Philippe Schmitt-Kopplin, Christoph Müller, Franz Bracher, Juliane Merl-Pham, Stefanie M Hauck, Michael Schloter, Ashley N Michael, Dayna Anderson, Linnea Honeker, Justyna Gozdz, Oksana Pivniouk, Carole Ober, Mark Holbreich, Fernando D Martinez, Shane A Snyder, Erika von Mutius, Donata Vercelli
Abstract
Background: Growing up on traditional European, and U.S. Amish, dairy farms, in close contact with cows and hay, protects children against asthma, and airway administration of extracts from dust collected from cowsheds of those farms prevents allergic asthma in mice.
Objectives: This study sought to begin identifying farm-derived asthma-protective agents.
Methods: Our work unfolded along two unbiased, independent but complementary discovery paths. Dust extracts from protective and non-protective farms (European and Amish cowsheds vs. European sheep sheds) were analyzed by comparative nuclear magnetic resonance fingerprinting and differential proteomics. Bioactivity-guided size fractionation focused on protective Amish cowshed dust extracts. Multiple in vitro and in vivo functional assays were used in both paths. Some of the proteins thus identified were characterized by in-solution and in-gel SDS-PAGE enzymatic digestion/peptide mapping followed by liquid chromatography/mass spectrometry. The cargo carried by these proteins was analyzed by untargeted liquid chromatography-high resolution mass spectrometry.
Results: Twelve carrier proteins of animal and plant origin, including the bovine lipocalins Bos d2 and odorant binding protein, were enriched in dust extracts from protective European cowsheds. A potent asthma-protective fraction of Amish cowshed dust extracts (≈0.5% of the total carbon content of unfractionated extracts) contained seven animal and plant proteins, including Bos d2 and odorant binding protein loaded with fatty acid metabolites from plants, bacteria and fungi.
Conclusions: Animals and plants from traditional farms produce proteins that transport hydrophobic microbial and plant metabolites. When delivered to mucosal surfaces, these agents might regulate airway responses.
背景：在传统的欧洲和美国阿米什族奶牛场长大，与奶牛和干草密切接触，可以保护儿童免受哮喘的侵袭，而通过气道给予从这些奶牛场牛棚收集的灰尘中提取的提取物可以预防小鼠的过敏性哮喘。
目的：本研究试图开始确定农场衍生的哮喘保护剂。
方法：我们的工作沿着两条无偏倚、独立但互补的发现路径展开。通过比较核磁共振指纹图谱和差异蛋白质组学分析了保护性和非保护性农场（欧洲和阿米什牛棚与欧洲羊棚）的灰尘提取物。以生物活性为导向的粒度分级主要针对保护性阿米什牛棚粉尘提取物。在这两种途径中都使用了多种体外和体内功能测定。通过溶液中和凝胶中SDS-PAGE酶促消化/肽图谱，然后通过液相色谱/质谱对由此鉴定的一些蛋白质进行表征。这些蛋白质携带的物质通过非靶向液相色谱-高分辨率质谱法进行分析。
结果：从欧洲防护牛舍的灰尘提取物中富集了12种动植物来源的载体蛋白，包括牛脂肪运载蛋白Bos d2和气味结合蛋白。阿米什牛棚粉尘提取物中一种有效的哮喘保护成分（约占未分级提取物总碳含量的0.5%）含有七种动植物蛋白，包括Bos d2和气味结合蛋白，该蛋白富含植物、细菌和真菌的脂肪酸代谢产物。
结论：传统农场的动植物产生的蛋白质可以运输疏水性微生物和植物代谢产物。当输送到黏膜表面时，这些药物可能会调节气道反应。
(中日友好医院呼吸与危重症医学科 顾宪民 摘译 林江涛 审校)
(J Allergy Clin Immunol. 2023 Jun 2;S0091-6749(23)00706-6. doi: 10.1016/j.jaci.2023.05.013.)
(J Allergy Clin Immunol. 2023 Jun 2;S0091-6749(23)00706-6. doi: 10.1016/j.jaci.2023.05.013.)
Asthma-protective agents in dust from traditional farm environments
Mauricius Marques Dos Santos, Vadim Pivniouk, Bettina Rankl, Alesia Walker, Giulia Pagani, Norbert Hertkorn, Philippe Schmitt-Kopplin, Christoph Müller, Franz Bracher, Juliane Merl-Pham, Stefanie M Hauck, Michael Schloter, Ashley N Michael, Dayna Anderson, Linnea Honeker, Justyna Gozdz, Oksana Pivniouk, Carole Ober, Mark Holbreich, Fernando D Martinez, Shane A Snyder, Erika von Mutius, Donata Vercelli
Abstract
Background: Growing up on traditional European, and U.S. Amish, dairy farms, in close contact with cows and hay, protects children against asthma, and airway administration of extracts from dust collected from cowsheds of those farms prevents allergic asthma in mice.
Objectives: This study sought to begin identifying farm-derived asthma-protective agents.
Methods: Our work unfolded along two unbiased, independent but complementary discovery paths. Dust extracts from protective and non-protective farms (European and Amish cowsheds vs. European sheep sheds) were analyzed by comparative nuclear magnetic resonance fingerprinting and differential proteomics. Bioactivity-guided size fractionation focused on protective Amish cowshed dust extracts. Multiple in vitro and in vivo functional assays were used in both paths. Some of the proteins thus identified were characterized by in-solution and in-gel SDS-PAGE enzymatic digestion/peptide mapping followed by liquid chromatography/mass spectrometry. The cargo carried by these proteins was analyzed by untargeted liquid chromatography-high resolution mass spectrometry.
Results: Twelve carrier proteins of animal and plant origin, including the bovine lipocalins Bos d2 and odorant binding protein, were enriched in dust extracts from protective European cowsheds. A potent asthma-protective fraction of Amish cowshed dust extracts (≈0.5% of the total carbon content of unfractionated extracts) contained seven animal and plant proteins, including Bos d2 and odorant binding protein loaded with fatty acid metabolites from plants, bacteria and fungi.
Conclusions: Animals and plants from traditional farms produce proteins that transport hydrophobic microbial and plant metabolites. When delivered to mucosal surfaces, these agents might regulate airway responses.
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不同的上皮先天免疫细胞转录回路是哮喘气道高反应性的基础
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