Red blood cell (RBC) transfusion remains a very common therapeutic in intensive care units, but is associated with unintended consequences in the critically ill population. While the lungs are a target organ, human lung injury is seldom caused by any single event, and RBC transfusion alone is generally not sufficient to induce injury but requires a host with increased susceptibility. Our ongoing interest in RBC transfusion and its potential role in modifying inflammatory responses have led us to focus upon the role of transfused oxidized, damaged RBC (oxRBC) and micro-particles bearing surface phosphatidylserine (PS) in altering the mononuclear phagocyte activation status and the role of host factors that protect from persistent lung inflammation and injury. We hypothesize that an endogenous countering mechanism exists whereby macrophage (M) engulfment of transfused oxRBC or PS+ micro-particles contained within standard RBC units results in suppression of macrophage activation through the release of the anti-inflammatory signal IL-10. This de-activating signal resulting from the ingestion of apoptotic bodies is required to curtail inflammation and promote the resolution phase of injury. Improper deactivation, through defective IL-10 signaling following ingestion of apoptotic bodies may increase the susceptibility of an individual to the risks of RBC transfusion. We have identified a novel murine model of the susceptible host and propose that mice deficient in thrombospondin-1 (TSP1), a multi-functional adhesive glycoprotein involved in phagocyte recognition of PS+ apoptotic bodies through the M scavenging receptor CD36, show defective IL-10 responses following LPS- induced lung inflammation and fail to resolve injury in addition to defective IL-10 signaling in th liver following transfusion of oxRBC. Based upon our findings, we propose an overall hypothesis that TSP1 functions as an extracellular, bridging molecule that mediates CD36-dependent IL-10 production, contributing to M deactivation necessary for proper resolution of lung inflammation following transfusion. When disrupted, the host is left vulnerable to the harmful effects of RBC transfusion and shows persistence of inflammation through impaired M deactivation. Utilizing human monocyte derived M (HMDM), murine M, tsp1-/-, and cd36-/- mice, we will determine whether (1) TSP1-mediated IL-10 production by M following engagement of RBC micro-particles or oxRBC requires CD36; (2) transfusion of stored RBC impairs resolution of lung inflammation induced by either LPS or bacterial pathogen in tsp1-/- mice through CD36-dependent IL-10 production; and (3) reconstitution of IL-10 hastens resolution of lung inflammation in tsp1-/- mice and that immune-modulatory properties of RBC micro-particles can be harnessed to reprogram mononuclear phagocytes with intact TSP- 1/CD36 axis toward resolution. Completion of these studies will identify a highly novel pathway underlying the immune-modulatory aspect of RBC transfusion and its contribution to lung inflammation and injury in the susceptible host.

Public Health Relevance

Red blood cell (RBC) transfusion remains a very common therapeutic in intensive care units, but is associated with increased risk of developing lung injur and worse outcome in the critically ill population. This proposal examines the role of host factor(s) that normally protect from persistent lung inflammation and injury, and when disrupted, the host is left vulnerable to the harmful effects of RBC transfusion. Completion of these studies will identify a highly novel pathway underlying the immune-modulatory aspect of RBC transfusion and its contribution to lung inflammation and injury in the susceptible host.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
4R01HL086884-09
Application #
9013491
Study Section
Lung Cellular, Molecular, and Immunobiology Study Section (LCMI)
Program Officer
Harabin, Andrea L
Project Start
2006-12-01
Project End
2018-03-31
Budget Start
2016-04-01
Budget End
2017-03-31
Support Year
9
Fiscal Year
2016
Total Cost
Indirect Cost
Name
University of Pittsburgh
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Bain, William; Lee, Janet S (2018) Ventilator Circuit Trash May Be a Research Treasure. Am J Respir Crit Care Med 197:979-980
Qu, Yanyan; Olonisakin, Tolani; Bain, William et al. (2018) Thrombospondin-1 protects against pathogen-induced lung injury by limiting extracellular matrix proteolysis. JCI Insight 3:
Lewis, Anthony J; Lee, Janet S; Rosengart, Matthew R (2018) Translational Sepsis Research: Spanning the Divide. Crit Care Med 46:1497-1505
Ryu, Hyunryul; Choi, Kyungyong; Qu, Yanyan et al. (2018) Label-free Neutrophil Enrichment from Patient-derived Airway Secretion Using Closed-loop Inertial Microfluidics. J Vis Exp :
Olonisakin, Tolani F; Lee, Janet S (2018) Fighting Bacterial Pathogens in the Lung: Platelets to the Rescue? Am J Respir Cell Mol Biol 58:282-283
Kitsios, Georgios D; Fitch, Adam; Manatakis, Dimitris V et al. (2018) Respiratory Microbiome Profiling for Etiologic Diagnosis of Pneumonia in Mechanically Ventilated Patients. Front Microbiol 9:1413
Lee, Janet S; Kim-Shapiro, Daniel B (2017) Stored blood: how old is too old? J Clin Invest 127:100-102
Ryu, Hyunryul; Choi, Kyungyong; Qu, Yanyan et al. (2017) Patient-Derived Airway Secretion Dissociation Technique To Isolate and Concentrate Immune Cells Using Closed-Loop Inertial Microfluidics. Anal Chem 89:5549-5556
Flitter, Becca A; Hvorecny, Kelli L; Ono, Emiko et al. (2017) Pseudomonas aeruginosa sabotages the generation of host proresolving lipid mediators. Proc Natl Acad Sci U S A 114:136-141
Han, SeungHye; Olonisakin, Tolani F; Pribis, John P et al. (2017) A checklist is associated with increased quality of reporting preclinical biomedical research: A systematic review. PLoS One 12:e0183591

Showing the most recent 10 out of 41 publications