Aeffner Famke, Woods Parker S, Davis Ian C
Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio.
Department of Veterinary Biosciences, The Ohio State University, Columbus, Ohio
Am J Physiol Lung Cell Mol Physiol. 2015 Dec 1;309(11):L1313-22. doi: 10.1152/ajplung.00130.2015. Epub 2015 Oct 2.
Extracellular nucleotides and nucleosides are important signaling molecules in the lung. Nucleotide and nucleoside concentrations in alveolar lining fluid are controlled by a complex network of surface ectonucleotidases. Previously, we demonstrated that influenza A/WSN/33 (H1N1) virus resulted in increased levels of the nucleotide ATP and the nucleoside adenosine in bronchoalveolar lavage fluid (BALF) of wild-type (WT) C57BL/6 mice. Influenza-induced acute lung injury (ALI) was highly attenuated in A1-adenosine receptor-knockout mice. Because AMP hydrolysis by the ecto-5'-nucleotidase (CD73) plays a central role in and is rate-limiting for generation of adenosine in the normal lung, we hypothesized that ALI would be attenuated in C57BL/6-congenic CD73-knockout (CD73-KO) mice. Infection-induced hypoxemia, bradycardia, viral replication, and bronchoconstriction were moderately increased in CD73-KO mice relative to WT controls. However, postinfection weight loss, pulmonary edema, and parenchymal dysfunction were not altered. Treatment of WT mice with the CD73 inhibitor 5'-(α,β-methylene) diphosphate (APCP) also had no effect on infection-induced pulmonary edema but modestly attenuated hypoxemia. BALF from CD73-KO and APCP-treated WT mice contained more IL-6 and CXCL-10/IFN-γ-induced protein 10, less CXCL-1/keratinocyte chemoattractant, and fewer neutrophils than BALF from untreated WT controls. BALF from APCP-treated WT mice also contained fewer alveolar macrophages and more transforming growth factor-β than BALF from untreated WT mice. These results indicate that CD73 is not necessary for development of ALI following influenza A virus infection and suggest that tissue-nonspecific alkaline phosphatase may be responsible for increased adenosine generation in the infected lung. However, they do suggest that CD73 has a previously unrecognized immunomodulatory role in influenza.
细胞外核苷酸和核苷是肺内重要的信号分子。肺泡衬液中的核苷酸和核苷浓度受表面外切核苷酸酶复杂网络的调控。此前,我们证明甲型流感病毒A/WSN/33(H1N1)可导致野生型(WT)C57BL/6小鼠支气管肺泡灌洗液(BALF)中核苷酸ATP和核苷腺苷水平升高。甲型流感病毒诱导的急性肺损伤(ALI)在A1-腺苷受体基因敲除小鼠中显著减轻。由于ecto-5'-核苷酸酶(CD73)对AMP的水解在正常肺中腺苷生成过程中起核心作用且是限速步骤,我们推测ALI在C57BL/6同源CD73基因敲除(CD73-KO)小鼠中会减轻。相对于WT对照,CD73-KO小鼠感染诱导的低氧血症、心动过缓、病毒复制和支气管收缩适度增加。然而,感染后体重减轻、肺水肿和实质功能障碍未改变。用CD73抑制剂5'-(α,β-亚甲基)二磷酸(APCP)处理WT小鼠对感染诱导的肺水肿也无影响,但可适度减轻低氧血症。与未处理的WT对照的BALF相比,CD73-KO和APCP处理的WT小鼠的BALF含有更多的IL-6和CXCL-10/IFN-γ诱导蛋白10,更少的CXCL-1/角质形成细胞趋化因子,以及更少的中性粒细胞。APCP处理的WT小鼠的BALF与未处理的WT小鼠的BALF相比,还含有更少的肺泡巨噬细胞和更多的转化生长因子-β。这些结果表明,CD73对于甲型流感病毒感染后ALI的发生并非必需,提示组织非特异性碱性磷酸酶可能是感染肺中腺苷生成增加的原因。然而,这些结果确实表明CD73在流感中具有此前未被认识到的免疫调节作用。