Evaluation of an interleukin-8 conjugated extracorporeal neutrophil reprogramming device for the treatment of acute inflammatory disorders
Year: 2016
Type: article
Abstract: Event Abstract Back to Event Evaluation of an interleukin-8 conjugated extracorporeal neutrophil reprogramming device for the treatment of acute inflammatory disorders Alexander Malkin1, 2, Kai Singbartl3, John A. Kellum2, 4 and William J. Federspiel1, 2, 4, 5 1 University of Pittsburgh, Department of Bioengineering, United States 2 University of Pittsburgh, McGowan Institute for Regenerative Medicine, United States 3 Penn State Hershey, Department of Anesthesiology, United States 4 University of Pittsburgh, Department of Critical Care Medicine, United States 5 University of Pittsburgh, Department of Chemical Engineering, United States Introduction: As one of the first responders to sites of inflammation, neutrophils play a critical role in the progression of acute inflammatory conditions such as sepsis. High quantities of neutrophils migrating to remote organs such as the lungs has been associated with increased morbidity and mortality by initiating multiple organ dysfunction or immune paralysis in septic patients. The migration and activation of neutrophils is regulated by CXCR-1 and CXCR-2, g-protein coupled receptors that bind to interleukin-8 (CXCL-8) [1]. It is hypothesized that exposure to high levels of immobilized CXCL-8 within an extracorporeal device will attenuate neutrophil migratory response. In this study, a method for immobilization of CXCL-8 on the inner lumen of AN69 dialyzer fibers was developed and constructed into a prototype extracorporeal device. Additionally, the device was evaluated by measuring CXCR-1 and CXCR-2 neutrophil surface receptor expression after human blood perfusion. Materials and Methods: First, amine modified AN69 dialyzer fibers (Gambro) were potted into a polycarbonate housing. Module ends were trimmed to open the fiber inner lumen. An NHS activated polyethylene-glycol spacer was immobilized on the inner lumen. CXCL-8 was then conjugated using NHS/EDC linkage. A control module used bovine serum albumin in place of CXCL-8. Fresh human whole blood was recirculated through the device for 90 minutes. A positive control was created by adding 1ug/ml soluble CXCL-8 to whole blood. Blood samples were collected at 45 and 90 minutes for analysis of CXCR-1 and CXCR-2 expression using flow cytometry. Results and Discussion: The soluble CXCL-8 positive control induced 24% and 94% downregulation of CXCR-1 and CXCR-2 respectively when compared to baseline blood, indicating complete migratory shutoff [2]. In the CXCL-8 modified module, neutrophil surface receptor expression of CXCR-1 and CXCR-2 decreased by 50% and 90%. CXCR-2 downregulation in this module closely tracked the positive control, indicating change in neutrophil migratory phenotype. The BSA modified control module induced less than 10% variation from baseline blood, demonstrating downregulation in the CXCL-8 module was due to the immobilized chemokine. Conclusion: An effective method for immobilization of CXCL-8 on the inner lumen of AN69 dialyzer fibers was developed. Furthermore, the CXCL-8 modified prototype module induced downregulation of CXCR-1 and CXCR-2 neutrophil surface receptors when perfused with human whole blood. Functionally, it is anticipated that CXCR-1 and CXCR-2 downregulation will reduce neutrophil migration and associated damage in acute inflammatory disorders. Further testing is being conducted to evaluate the functional response of reprogrammed neutrophils to a chemotactic gradient. Gambro Industries for supplying amine functionalized dialyzer fibersReferences:[1] Nasser, M. W. et al. CXCR1 and CXCR2 Activation and Regulation Role of Aspartate 199 of the Second Extracellular Loop of CXCR2 in CXCL8-Mediated Rapid Receptor Internalization. J. Biol. Chem. 282, 6906–6915 (2007).[2] Feniger-Barish, R., Yron, I., Meshel, T., Matityahu, E. & Ben-Baruch, A. IL-8-Induced Migratory Responses through CXCR1 and CXCR2: Association with Phosphorylation and Cellular Redistribution of Focal Adhesion Kinase†. Biochemistry (Mosc.) 42, 2874–2886 (2003). Keywords: Clinical relevance, device, biofunctionalization, Cell modulation Conference: 10th World Biomaterials Congress, Montréal, Canada, 17 May - 22 May, 2016. Presentation Type: Poster Topic: Biomaterials for cardiovascular applications, vascular grafts and embolic devices Citation: Malkin A, Singbartl K, Kellum JA and Federspiel WJ (2016). Evaluation of an interleukin-8 conjugated extracorporeal neutrophil reprogramming device for the treatment of acute inflammatory disorders. Front. Bioeng. Biotechnol. Conference Abstract: 10th World Biomaterials Congress. doi: 10.3389/conf.FBIOE.2016.01.02192 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Mar 2016; Published Online: 30 Mar 2016. Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Alexander Malkin Kai Singbartl John A Kellum William J Federspiel Google Alexander Malkin Kai Singbartl John A Kellum William J Federspiel Google Scholar Alexander Malkin Kai Singbartl John A Kellum William J Federspiel PubMed Alexander Malkin Kai Singbartl John A Kellum William J Federspiel Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.
Language: English
Subfield: Immunology
Field: Immunology and Microbiology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $3,801
OpenAlex ID: W2332005397