Leukocyte compartments in the mouse lung: distinguishing between marginated, interstitial, and alveolar cells in response to injury.
Leukocyte compartments in the mouse lung: distinguishing between marginated, interstitial, and alveolar cells in response to injury.
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DOI：
10.1016/j.jim.2011.09.013
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发表时间：
2012-01-31
影响因子：
2.2
通讯作者：
Linden, Joel
中科院分区：
文献类型：
--
作者：
Barletta, Kathryn E.;Cagnina, R. Elaine;Wallace, Kori L.;Ramos, Susan I.;Mehrad, Borna;Linden, Joel
我们开发了一种基于流式细胞术的检测方法，可以同时定量小鼠肺边缘血管、间质和肺泡室中的多个白细胞群。静脉注射一种荧光标记的抗cd45抗体用于标记循环和边缘血管白细胞。在血管冲洗去除非粘附细胞和收集支气管肺泡灌洗液（BAL）后，对肺进行消化，并在体外添加第二种抗cd45荧光抗体以识别不在血管间隙的细胞。在naïve小鼠肺中，我们发现了大约1100万个CD45+白细胞，其中87%（950万）位于血管边缘室，由17%的NK细胞，17%的中性粒细胞，57%的单核髓样细胞（单核细胞，巨噬细胞前体和树突状细胞）和10%的T细胞（CD4+， CD8+和不变的NKT细胞）组成。包括间质室在内的非血管室含有7.7 × 105个细胞，其中NK细胞占49%，树突状细胞占25%，其他单核髓细胞占16%。肺泡间室绝大多数为巨噬细胞（5.63 × 105，占93%）。接下来，我们研究了胃酸损伤后白细胞边缘和外渗到肺内的情况。损伤后1小时，血液中中性粒细胞明显升高，而所有其他循环白细胞平均下降79%。损伤后4小时，边缘中性粒细胞、NK细胞、CD4+和CD8+ T细胞数量达到峰值，肺泡NK细胞数量达到峰值。大多数间质细胞由dc、中性粒细胞和CD4+ T细胞组成，大多数肺泡间室细胞由巨噬细胞、中性粒细胞和NK细胞组成。损伤后24小时，所有边缘和间质白细胞数量下降，肺泡中性粒细胞达到峰值。总之，我们已经开发了一种新的检测方法来研究肺部炎症后的白细胞边缘和运输，并表明边缘细胞占肺白细胞的很大一部分，在肺损伤后不久增加。该试验可能会对未来的研究产生兴趣，以确定白细胞是否在粘附内皮细胞时被激活，并具有将其与间质细胞和循环细胞区分开来的特性。
We developed a flow cytometry-based assay to simultaneously quantify multiple leukocyte populations in the marginated vascular, interstitial, and alveolar compartments of the mouse lung. An intravenous injection of a fluorescently labeled anti-CD45 antibody was used to label circulating and marginated vascular leukocytes. Following vascular flushing to remove non-adherent cells and collection of broncho-alveolar lavage (BAL) fluid, lungs were digested and a second fluorescent anti-CD45 antibody was added ex vivo to identify cells not located in the vascular space. In the naïve mouse lung, we found about 11 million CD45+ leukocytes, of which 87% (9.5 million) were in the vascular marginated compartment, consisting of 17% NK cells, 17% neutrophils, 57% mononuclear myeloid cells (monocytes, macrophage precursors and dendritic cells), and 10% T cells (CD4+, CD8+, and invariant NKT cells). Non-vascular compartments including the interstitial compartment contained 7.7 × 105 cells, consisting of 49% NK cells, 25% dendritic cells, and 16% other mononuclear myeloid cells. The alveolar compartment was overwhelmingly populated by macrophages (5.63 × 105 cells, or 93%). We next studied leukocyte margination and extravasation into the lung following acid injury, a model of gastric aspiration. At 1 hour after injury, neutrophils were markedly elevated in the blood while all other circulating leukocytes declined by an average of 79 percent. At 4 hours after injury, there was a peak in the numbers of marginated neutrophils, NK cells, CD4+ and CD8+ T cells and a peak in the number of alveolar NK cells. Most interstitial cells consisted of DCs, neutrophils, and CD4+ T cells, and most alveolar compartment cells consisted of macrophages, neutrophils, and NK cells. At 24 hours after injury, there was a decline in the number of all marginated and interstitial leukocytes and a peak in alveolar neutrophils. In sum, we have developed a novel assay to study leukocyte margination and trafficking following pulmonary inflammation and show that marginated cells comprise a large fraction of lung leukocytes that increases shortly after lung injury. This assay may be of interest in future studies to determine if leukocytes become activated upon adherence to the endothelium, and have properties that distinguish them from interstitial and circulating cells.
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影响因子：
3.1
作者：
Wang, EJ;Ouellet, N;Bergeron, MG
通讯作者：
Bergeron, MG
DOI：
10.1164/rccm.200602-256oc
发表时间：
2007-06-01
影响因子：
24.7
作者：
Phadke, Anagha P.;Akangire, Gangaram;Mehrad, Borna
通讯作者：
Mehrad, Borna
影响因子：
64.8
作者：
Imai Y;Kuba K;Rao S;Huan Y;Guo F;Guan B;Yang P;Sarao R;Wada T;Leong-Poi H;Crackower MA;Fukamizu A;Hui CC;Hein L;Uhlig S;Slutsky AS;Jiang C;Penninger JM
通讯作者：
Penninger JM
DOI：
10.4049/jimmunol.181.8.5183
发表时间：
2008-10-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
von Vietinghoff S;Ley K
通讯作者：
Ley K
影响因子：
8.8
作者：
KNIGHT, PR;DRUSKOVICH, G;JOHNSON, KJ
通讯作者：
JOHNSON, KJ