Venous Thrombosis–Associated Inflammation and Attenuation With Neutralizing Antibodies to Cytokines and Adhesion Molecules

Abstract

Thrombosis and inflammation are closely related. However, the response of the vein wall to venous thrombosis has been poorly documented. This study examines the hypothesis that venous thrombosis is associated with an inflammatory response in the vein wall. In a rat model of inferior vena caval thrombosis, vein wall was temporally examined for inflammation by assessment of histopathology, leukocyte morphometrics, and cytokine levels. Animals were killed 1 hour and 1, 3, and 6 days after thrombus induction. Our findings demonstrated an early (day 1) neutrophil infiltration into the vein wall followed by a later (days 3 and 6) monocyte/macrophage and lymphocyte response. Cytokines were elevated only under conditions of venous thrombosis. Levels of epithelial neutrophil activating protein-78 (ENA-78), tumor necrosis factor-alpha (TNF), interleukin-6, and JE/monocyte chemoattractant protein-1 (JE/MCP-1) increased over the 6-day period, while macrophage inflammatory protein-1 alpha (MIP-1 alpha) peaked at day 3 after thrombus induction. Additionally, rats were passively immunized with neutralizing antibodies to TNF, ENA-78, MIP-1 alpha, JE/MCP-1, intercellular adhesion molecule-1 (ICAM-1), and CD18 compared with control antibodies. The most effective antibody early after thrombus induction for attenuating vein wall neutrophil extravasation was anti-TNF (P < .01). The monocyte/macrophage extravasation was inhibited most by anti-ICAM-1 followed by anti-TNF (P < .01). These findings demonstrate that venous thrombosis is associated with significant vein wall inflammation that is partially inhibited by neutralizing antibodies to cytokines and adhesion molecules.

Targeted Deletion of CCR2 Impairs Deep Vein Thombosis Resolution in a Mouse Model

“…Whereas other studies have suggested the early DVT environment is proinflammatory (4,5,8), the data herein support several mechanisms associated with CCR2 signaling activity that directly affects thrombus resolution (Fig. 6).…”

“…Although four main chemokine families exist, the ones most relevant to DVT resolution are the cysteine-cysteine (CC) and cysteine-X-cysteine (CXC) chemokine types. For example, both CXC and CC chemokines are present within a resolving thrombus (4), and exogenous administration of IL-8 (CXC chemokine) and MCP-1 (a CC chemokine) have been shown to accelerate thrombus resolution in experimental models (7,8). Many pathophysiological processes involve CCR2 signaling, including peritoneal and pulmonary inflammation, and atherogenesis (9 -11).…”

“…The rodent model of stasis-induced DVT has been well described (4,5,7,30) and used B6/129 (wild-type (WT) controls, F2 hybrids; The Jackson Laboraory) and B6/129 CCR2 Ϫ/Ϫ mice (10) weighing 20 -30 g, between 4 and 6 wk old. The mice underwent general inhalational anesthesia and, via a laparotomy, the inferior vena cava (IVC) was ligated with a 6-0 polypropylene suture.…”

“…Quantification of peptide mediators was normalized to total protein in the sample, by a modified Bradford assay per manufacturer's instructions (Pierce) with serial dilutions of BSA (Sigma-Aldrich) as standards. Tissue homogenate ELISAs for mouse MIP-1␣, RANTES, MCP-1, TGF␤, IFN-␥, IL-2, IL-4, IL-5, and IL-13 were performed with speciesspecific primary Abs quantified using a double ligand technique, as has been described for similar chemokines (4,5,35). Mouse bFGF and VEGF were analyzed by using a commercial ELISA according to the manufacturer's instructions (R&D Systems).…”

Correspondence of ultrasound elasticity imaging to direct mechanical measurement in aging DVT in rats

“…Animal studies were performed using a rat model of stasis induced venous thrombosis (Wakefield et al 1995). The protocol was approved by the University of Michigan Committee on the Use and Care of Animals and strictly complied with the National Institutes of Health Guide for the Care and Use of Laboratory Animals.…”