High‐level expression of cytokine‐induced neutrophil chemoattractant (CINC) by a metastatic rat cell line: Purification and production of blocking antibodies

“…The production and purification of polyclonal goat anti-rat CINC-1 Abs was reproduced precisely as previously published (31). Briefly, we expressed recombinant CINC-1 using a previously described CINC-1 expression vector (pJZ240) that fused the N terminus of CINC-1 to the C terminus of the gene for Schistosoma japonicum GST in the expression vector pGEX-3ϫ (Pharmacia/GE Health Sciences).…”

Section: Preparation Of Anti-cinc-1 Polyclonal Antibodiesmentioning

confidence: 99%

Inhibition of CINC-1 Decreases Right Ventricular Damage Caused by Experimental Pulmonary Embolism in Rats

Zagorski

Gellar

Obraztsova

et al. 2007

The Journal of Immunology

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Right ventricular (RV) dysfunction is a strong risk factor for poor clinical outcome following pulmonary embolism (PE), the third most prevalent cardiovascular disease. Previous studies in our laboratory demonstrated that RV failure during PE is mediated, in part, by neutrophil-dependant cardiac inflammation. In this study we use DNA microarray analysis of gene expression to demonstrate that PE results in increased expression of the CXC chemokines CINC-1 and CINC-2 between 6 and 18 h after the start of PE in a rat model of PE. Neutrophils accumulate in RV tissue by 18 h, and this inflammation is associated with decreased right heart function. Treatment of rats with Abs to CINC-1 significantly suppressed neutrophil accumulation in RVs during PE (52% reduction in tissue myeloperoxidase) and ameliorated RV failure. In addition, plasma concentration of cardiac troponin I, an established diagnostic marker for cardiac damage, was reduced by 90%. These results suggest that selective anti-inflammatory therapies targeted at neutrophil chemoattractants will reduce cardiac inflammation and reduce RV damage in the setting of PE.

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“…Neutrophils were purified using Polymorphprep cell separation medium (Accurate Chemical and Scientific, Westbury, NY) and chemotaxis assays done using 24-well Corning-Costar Transwell migration chambers as previously described (40,45) but with a modified cell counting procedure. Formaldehyde/EDTA-fixed cells were collected by centrifugation (5 min at 450 ϫ g) and counted with a hemacytometer.…”

Section: Chemotaxis Assaysmentioning

confidence: 99%

“…To test this, we used anti-CINC (40,45) and anti-MIP-2 (Santa Cruz Biotechnology, Santa Cruz CA) Abs to block the neutrophil chemotactic activity in a pool of PE2.0 BALs in vitro. The BAL pool was diluted 1:3 with chemotaxis buffer before use.…”

Section: Chemokines Contribute To the Chemotactic Activity In Pe2mentioning

confidence: 99%

Chemokines Accumulate in the Lungs of Rats with Severe Pulmonary Embolism Induced by Polystyrene Microspheres

Zagorski

Debelak

Gellar

et al. 2003

The Journal of Immunology

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Pulmonary thromboembolism (PEm) is a serious and life threatening disease and the most common cause of acute pulmonary vascular occlusion. Even following successful treatment of PEm, many patients experience long-term disability due to diminished heart and lung function. Considerable damage to the lungs presumably occurs due to reperfusion injury following anti-occlusive treatments for PEm and the resulting chronic inflammatory state in the lung vasculature. We have used a rat model of irreversible PEm to ask whether pulmonary vascular occlusion in the absence of reperfusion is itself sufficient to induce an inflammatory response in lungs. By adjusting the severity of the vascular occlusion, we were able to generate hypertensive and nonhypertensive PEm, and then examine lung tissue for expression of CXC and C-C chemokine genes and bronchoalveolar lavage (BAL) fluid for the presence of chemokine proteins. Hypertensive and nonhypertensive PEm resulted in increased expression of both CXC and C-C chemokines genes in lung tissues. Hypertensive PEm was also associated with a 50–100-fold increase in protein content in lung BAL fluid, which included the CXC chemokines cytokine-induced neutrophil chemoattractant and macrophage-inflammatory protein 2. The presence of chemokines in BALs was reflected by a potent neutrophil chemotactic activity in in vitro chemotaxis assays. Abs to cytokine-induced neutrophil chemoattractant blocked the in vitro neutrophil chemotactic activity of BAL by 44%. Our results indicate that the ischemia and hypertension associated with PEm are sufficient to induce expression of proinflammatory mediators such as chemokines, and establish a proinflammatory environment in the ischemic lung even before reperfusion.

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High‐level expression of cytokine‐induced neutrophil chemoattractant (CINC) by a metastatic rat cell line: Purification and production of blocking antibodies

Cited by 44 publications

References 20 publications

Angiotensin II Induces Neutrophil Accumulation In Vivo Through Generation and Release of CXC Chemokines

Angiotensin II Induces Neutrophil Accumulation In Vivo Through Generation and Release of CXC Chemokines

Inhibition of CINC-1 Decreases Right Ventricular Damage Caused by Experimental Pulmonary Embolism in Rats

Chemokines Accumulate in the Lungs of Rats with Severe Pulmonary Embolism Induced by Polystyrene Microspheres

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