Quantitative assessment of macrophage functions in repair and fibrosis.
Quantitative assessment of macrophage functions in repair and fibrosis.
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DOI：
10.1002/0471142735.im1422s93
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发表时间：
2011-04
影响因子：
--
通讯作者：
Ramalingam, Thirumalai
中科院分区：
文献类型：
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作者：
Wynn, Thomas A;Barron, Luke;Thompson, Robert W;Madala, Satish K;Wilson, Mark S;Cheever, Allen W;Ramalingam, Thirumalai
巨噬细胞通过调节细胞外基质的代谢，在创伤修复和纤维化中发挥关键作用。巨噬细胞本身可以处理基质成分，但也可以招募和改变其他类型细胞的功能，这些细胞直接构建或降解细胞外基质。经典激活的巨噬细胞(CAM，又称M1)倾向于促进组织损伤，而交替激活的巨噬细胞(AAM，又称M2)往往与创伤修复和纤维化的机制有关。然而，最近的研究表明，表达精氨酸酶-1的AAM通过抑制抗原特异性T细胞反应来抑制慢性炎症和纤维化，而不是促进胶原沉积。本单元描述了测量巨噬细胞和整个组织中精氨酸酶活性的方法，以及量化AAM的T细胞抑制活性的方法。还描述了可用于定量组织和支气管肺泡灌洗液中胶原蛋白水平的改良羟脯氨酸和可溶性胶原测定法。本单元中的方案应为研究人员提供所有必要的信息，以测量精氨酸酶活性，并将观察到的活性与纤维化的进展和缓解相关联。
Macrophages play key roles in wound repair and fibrosis by regulating extracellular matrix turnover. Macrophages can process matrix components themselves, but also recruit and alter the functions of other cell types that directly build or degrade extracellular matrix. Classically activated macrophages (CAM, also called M1) tend to promote tissue injury while alternatively activated macrophages (AAM, also called M2) are often linked with the mechanisms of wound repair and fibrosis. However, rather than promoting collagen deposition, recent studies suggest that arginase-1-expressing AAM suppress chronic inflammation and fibrosis by inhibiting antigen-specific T cell responses. This unit describes methods to measure arginase activity in macrophages and whole tissues as well as assays to quantify the T cell suppressive activity of AAMs. Modified hydroxyproline and soluble collagen assays that can be used to quantify collagen levels in tissues and brochoalveolar lavage fluid are also described. The protocols in this unit should provide the investigator with all the necessary information required to measure arginase activity and to correlate the observed activity with the progression and resolution of fibrosis.