PDF《Pulmonary Stromal-Derived Factor-1》

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3 Abbreviations used in this paper: ALI, acute lung injury;

BAL, bronchoalveolar lavage;

IHC, immunohistochemistry;

SDF-1, stromal-derived factor-1. Copyright ?

2007 by The American Association of Immunologists, Inc. 0022-1767/07/$2.00 The Journal of Immunology www.jimmunol.org by guest on May 30,

2019 http://www.jimmunol.org/ Downloaded from lung ?brosis (34, 36), its expression has not been examined in the setting of ALI. Our group and others have previously demonstrated that SDF-1 is a chemoattractant for neutrophils within the bone marrow and is responsible for marrow retention and marrow homing of these cells under homeostatic conditions (29, 38C40). How the CXCR4/ SDF-1 axis might participate in the development of in?ammation is poorly understood, but several reports support a role for this pathway in the tissue migration of leukocytes, including neutro- phils, in animal models of arthritis (41C44) and peritonitis (45). A role for SDF-1 in lung in?ammation has been suggested by animal models of asthma in which a CXCR4 blockade attenuates both the lymphocyte and eosinophil responses and reduces airway hyper- reactivity (46, 47). However, no investigations of the CXCR4/ SDF-1 axis in the recruitment of neutrophils to the lung have been reported, and whether pulmonary SDF-1 expression occurs during the development and progression of ALI is unknown. We inves- tigated pulmonary SDF-1 expression during ALI in mice and hu- mans, and examined the potential role of SDF-1 in neutrophil re- cruitment using a mouse model of pneumonitis. Materials and Methods Mice Four- to eight-week-old female C57BL/6 mice were obtained from Harlan and housed in the animal facilities of the University of Vermont College of Medicine. All experiments were performed in accordance with the Animal Welfare Act and the U.S. Public Health Service Policy on Humane Care and Use of Laboratory Animals after review of the protocol by the Animal Care and Use Committee of the University of Vermont (Burlington, VT). Reagents Mouse anti-human SDF-1 mAb (clone 79018;

R&

D Systems), rabbit poly- clonal anti-cytokeratin (catalog no. 18-0059, Zymed Laboratories), Alexa Fluor 568-labeled goat anti-mouse F(ab?)2 (catalog no. A11019;

Invitrogen Life Technologies), and Alexa Fluor 488-labeled donkey anti-rabbit IgG (catalog no. A21206;

Invitrogen Life Technologies) were purchased for use in immunohistology. Biotinylated rat anti-mouse CXCR4 mAb (2B11/ CXCR4) and control (rat IgG2b) were obtained from BD Pharmingen for use in ?ow cytometry. Rat anti-mouse CD16/CD32 and FITC-conjugated rat anti-mouse Ly-6G (Gr-1) mAbs were ........