UNLABELLED: Proinflammatory activation of vascular endothelium leading to increased surface expression of adhesion molecules and neutrophil (PMN) sequestration and subsequent activation is paramount in the development of acute lung injury and organ injury in injured patients. We hypothesize that α-enolase, which accumulates in injured patients, primes PMNs and causes proinflammatory activation of endothelial cells leading to PMN-mediated cytotoxicity. METHODS: Proteomic analyses of field plasma samples from injured versus healthy patients were used for protein identification. Human pulmonary microvascular endothelial cells (HMVECs) were incubated with α-enolase or thrombin, and intercellular adhesion molecule-1 surface expression was measured by flow cytometry. A two-event in vitro model of PMN cytotoxicity HMVECs activated with α-enolase, thrombin, or buffer was used as targets for lysophosphatidylcholine-primed or buffer-treated PMNs. The PMN priming activity of α-enolase was completed, and lysates from both PMNs and HMVECs were immunoblotted for protease-activated receptor 1 (PAR-1) and PAR-2 and coprecipitation of α-enolase with PAR-2 and plasminogen/plasmin. RESULTS: α-Enolase increased 10.8-fold in injured patients (P < 0.05). Thrombin and α-enolase significantly increased intercellular adhesion molecule-1 surface expression on HMVECs, which was inhibited by antiproteases, induced PMN adherence, and served as the first event in the two-event model of PMN cytotoxicity. α-Enolase coprecipitated with PAR-2 and plasminogen/plasmin on HMVECs and PMNs and induced PMN priming, which was inhibited by tranexamic acid, and enzymatic activity was not required. CONCLUSIONS: α-Enolase increases after injury and may activate pulmonary endothelial cells and prime PMNs through plasmin activity and PAR-2 activation. Such proinflammatory endothelial activation may predispose to PMN-mediated organ injury.
UNLABELLED: Proinflammatory activation of vascular endothelium leading to increased surface expression of adhesion molecules and neutrophil (PMN) sequestration and subsequent activation is paramount in the development of acute lung injury and organ injury in injured patients. We hypothesize that α-enolase, which accumulates in injured patients, primes PMNs and causes proinflammatory activation of endothelial cells leading to PMN-mediated cytotoxicity. METHODS: Proteomic analyses of field plasma samples from injured versus healthy patients were used for protein identification. Human pulmonary microvascular endothelial cells (HMVECs) were incubated with α-enolase or thrombin, and intercellular adhesion molecule-1 surface expression was measured by flow cytometry. A two-event in vitro model of PMN cytotoxicity HMVECs activated with α-enolase, thrombin, or buffer was used as targets for lysophosphatidylcholine-primed or buffer-treated PMNs. The PMN priming activity of α-enolase was completed, and lysates from both PMNs and HMVECs were immunoblotted for protease-activated receptor 1 (PAR-1) and PAR-2 and coprecipitation of α-enolase with PAR-2 and plasminogen/plasmin. RESULTS: α-Enolase increased 10.8-fold in injured patients (P < 0.05). Thrombin and α-enolase significantly increased intercellular adhesion molecule-1 surface expression on HMVECs, which was inhibited by antiproteases, induced PMN adherence, and served as the first event in the two-event model of PMN cytotoxicity. α-Enolase coprecipitated with PAR-2 and plasminogen/plasmin on HMVECs and PMNs and induced PMN priming, which was inhibited by tranexamic acid, and enzymatic activity was not required. CONCLUSIONS: α-Enolase increases after injury and may activate pulmonary endothelial cells and prime PMNs through plasmin activity and PAR-2 activation. Such proinflammatory endothelial activation may predispose to PMN-mediated organ injury.
Authors: Martin Steinhoff; Jörg Buddenkotte; Victoria Shpacovitch; Anke Rattenholl; Corinna Moormann; Nathalie Vergnolle; Thomas A Luger; Morley D Hollenberg Journal: Endocr Rev Date: 2005-02 Impact factor: 19.871
Authors: Angelo D'Alessandro; Monika Dzieciatkowska; Erik D Peltz; Ernest E Moore; Janeen R Jordan; Christopher C Silliman; Anirban Banerjee; Kirk C Hansen Journal: Shock Date: 2014-12 Impact factor: 3.454
Authors: M Dzieciatkowska; C C Silliman; E E Moore; M R Kelher; A Banerjee; K J Land; M Ellison; F B West; D R Ambruso; K C Hansen Journal: Vox Sang Date: 2013-05-11 Impact factor: 2.144
Authors: Lakmali Munasinghage Silva; Andrew Gary Lum; Collin Tran; Molly W Shaw; Zhen Gao; Matthew J Flick; Niki M Moutsopoulos; Thomas H Bugge; Eric S Mullins Journal: Blood Date: 2019-05-17 Impact factor: 22.113
Authors: Nathan Clendenen; Geoffrey R Nunns; Ernest E Moore; Julie A Reisz; Eduardo Gonzalez; Erik Peltz; Christopher C Silliman; Miguel Fragoso; Travis Nemkov; Matthew J Wither; Kirk Hansen; Anirban Banerjee; Hunter B Moore; Angelo DʼAlessandro Journal: J Trauma Acute Care Surg Date: 2017-10 Impact factor: 3.313
Authors: Alexander P Morton; Jamie B Hadley; Arsen Ghasabyan; Marguerite R Kelher; Ernest E Moore; Shaun Bevers; Monika Dzieciatkowska; Kirk C Hansen; Mitchell S Cohen; Anirban Banerjee; Christopher C Silliman Journal: J Trauma Acute Care Surg Date: 2022-01-01 Impact factor: 3.697
Authors: Christopher C Silliman; Marguerite R Kelher; Samina Y Khan; F Bernadette West; Nathan J D McLaughlin; David J Elzi; Kelly England; Jason Bjornsen; Susan A Kuldanek; Anirban Banerjee Journal: Transfusion Date: 2017-09-06 Impact factor: 3.157