Department of Clinical Sciences, Malmö, Section for Surgery, Lund University, Lund, Sweden.
Infect Immun. 2013 Jul;81(7):2499-506. doi: 10.1128/IAI.00126-13. Epub 2013 Apr 29.
T-cell dysfunction increases susceptibility to infections in patients with sepsis. In the present study, we hypothesized that Rho kinase signaling might regulate induction of T-cell dysfunction in abdominal sepsis. Male C57BL/6 mice were treated with the specific Rho kinase inhibitor Y-27632 (5 mg/kg of body weight) prior to cecal ligation and puncture (CLP). Spleen CD4 T-cell apoptosis, proliferation, and percentage of regulatory T cells (CD4(+) CD25(+) Foxp3(+)) were determined by flow cytometry. Formation of gamma interferon (IFN-γ) and interleukin 4 (IL-4) in the spleen and plasma levels of HMBG1, IL-17, and IL-6 were quantified by use of enzyme-linked immunosorbent assay (ELISA). It was found that CLP evoked apoptosis and decreased proliferation in splenic CD4 T cells. Inhibition of Rho kinase activity decreased apoptosis and enhanced proliferation of CD4 T cells in septic animals. In addition, CLP-evoked induction of regulatory T cells in the spleen was abolished by Rho kinase inhibition. CLP reduced the levels of IFN-γ and IL-4 in the spleen. Pretreatment with Y-27632 inhibited the sepsis-induced decrease in IFN-γ but not IL-4 formation in the spleen. CLP increased plasma levels of high-mobility group box 1 (HMGB1) by 20-fold and IL-6 by 19-fold. Inhibition of Rho kinase decreased this CLP-evoked increase of HMGB1, IL-6, and IL-17 levels in the plasma by more than 60%, suggesting that Rho kinase regulates systemic inflammation in sepsis. Moreover, we observed that pretreatment with Y-27632 abolished CLP-induced bacteremia. Together, our novel findings indicate that Rho kinase is a powerful regulator of T-cell immune dysfunction in abdominal sepsis. Thus, targeting Rho kinase signaling might be a useful strategy to improve T-cell immunity in patients with abdominal sepsis.
T 细胞功能障碍会增加脓毒症患者感染的易感性。在本研究中,我们假设 Rho 激酶信号可能调节腹部脓毒症中 T 细胞功能障碍的诱导。雄性 C57BL/6 小鼠在盲肠结扎和穿刺(CLP)前用特异性 Rho 激酶抑制剂 Y-27632(5mg/kg 体重)处理。通过流式细胞术测定脾 CD4 T 细胞凋亡、增殖和调节性 T 细胞(CD4+CD25+Foxp3+)的百分比。通过酶联免疫吸附试验(ELISA)定量测定脾和血浆中γ干扰素(IFN-γ)和白细胞介素 4(IL-4)的形成以及高迁移率族蛋白 B1(HMBG1)、白细胞介素 17(IL-17)和白细胞介素 6(IL-6)的血浆水平。结果发现,CLP 可引起脾 CD4 T 细胞凋亡并降低增殖。Rho 激酶活性抑制可减少脓毒症动物 CD4 T 细胞的凋亡并增强其增殖。此外,Rho 激酶抑制可消除 CLP 诱导的脾调节性 T 细胞的诱导。CLP 降低脾中 IFN-γ和 IL-4 的水平。Y-27632 预处理抑制了脓毒症引起的脾 IFN-γ形成减少,但不影响 IL-4 的形成。CLP 使高迁移率族蛋白 B1(HMGB1)和白细胞介素 6(IL-6)的血浆水平增加了 20 倍,白细胞介素 17(IL-17)增加了 19 倍。Rho 激酶抑制可使 HMGB1、IL-6 和 IL-17 水平在血浆中的 CLP 诱导增加降低 60%以上,表明 Rho 激酶调节脓毒症中的全身炎症。此外,我们观察到 Y-27632 预处理可消除 CLP 诱导的菌血症。总之,我们的新发现表明 Rho 激酶是腹部脓毒症中 T 细胞免疫功能障碍的强大调节剂。因此,靶向 Rho 激酶信号可能是改善腹部脓毒症患者 T 细胞免疫的有效策略。