The mechanical property of neutrophils to inflammatory process and sequestration into the lung.

2004 Fiscal Year Final Research Report Summary

• Project/Area Number: 15591912
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Emergency medicine
• Research Institution: Akita University
• Principal Investigator: SAITO Hajime Akita University, School of Medicine, Assistant Professor, 医学部, 講師 (20323149)
• Co-Investigator(Kenkyū-buntansha): OGAWA Jun-ichi Akita University, School of Medicine, Professor, 医学部, 教授 (20112774) ; MINAMIYA Yoshihiro Akita University, School of Medicine, Associate Professor, 医学部, 助教授 (30239321)
• Project Period (FY): 2003 – 2004
• Keywords: neutrophils / transmigration / sequestration / lung injury / deformability / actin / bone marrow release / antithrombin

Research Abstract

Because the spherical diameter of pulmonary capillaries is smaller than that of neutrophils, increased neutrophil stiffness or, conversely, decreased neutrophil deformability is a key step in the initial sequestration of neutrophils within the lungs during inflammatory processes. Antithrombin III (AT) is known exert a therapeutic effect against disseminated intravascular coagulation, and accumulating evidence suggests that AT also has anti-inflammatory properties. The mechanisms of its anti-inflammatory effects remain unclear, but in a rat endotoxin model AT apparently inhibited neutrophil sequestration in the lung. In the present in vitro study, therefore, we examined the effect of AT on the deformability of human neutrophils and correlated those findings with their F-actin content. Isolated human neutrophils were stimulated with fMLP (1 μM, 2 min) in the presence or absence of the α, β or low-heparin-affinity isoforms of AT (1 IU/ml, 20 min), and deformability was evaluated using a filter assay system. Neutrophils were also stained with FITC-phalloidin and subjected to FACS-scan to assess F-actin content. The results showed that pretreatment with any of the three AT isoforms similarly inhibited the decreased neutrophil deformability and increased F-actin content. Notably, heparinase had no effect on either deformability or F-actin content in the presence or absence of AT, which was somewhat unexpected, as heparin sulfate proteoglycans likely function as AT receptors. These findings suggested that AT inhibits the increase in neutrophil stiffness seen during inflammatory processes by inhibiting actin polymerization via a heparin-independent pathway.