Myocardial ischemia/reperfusion injury is mediated by leukocytic toll-like receptor-2 and reduced by systemic administration of a novel anti-toll-like receptor-2 antibody.
BACKGROUND
Reperfusion therapy for myocardial infarction is hampered by detrimental inflammatory responses partly via Toll-like receptor (TLR) activation. Targeting TLR signaling may optimize reperfusion therapy and enhance cell survival and heart function after myocardial infarction. Here, we evaluated the role of TLR2 as a therapeutic target using a novel monoclonal anti-TLR2 antibody.


RESULTS
Mice underwent 30 minutes of ischemia followed by reperfusion. Compounds were administered 5 minutes before reperfusion. Cardiac function and dimensions were assessed at baseline and 28 days after infarction with 9.4-T mouse magnetic resonance imaging. Saline and IgG isotype treatment resulted in 34.5 + or - 3.3% and 31.4 + or - 2.7% infarction, respectively. Bone marrow transplantation experiments between wild-type and TLR2-null mice revealed that final infarct size is determined by circulating TLR2 expression. A single intravenous bolus injection of anti-TLR2 antibody reduced infarct size to 18.9 + or - 2.2% (P=0.001). Compared with saline-treated mice, anti-TLR2-treated mice exhibited less expansive remodeling (end-diastolic volume 68.2 + or - 2.5 versus 76.8 + or - 3.5 microL; P=0.046) and preserved systolic performance (ejection fraction 51.0 + or - 2.1% versus 39.9 + or - 2.2%, P=0.009; systolic wall thickening 3.3 + or - 6.0% versus 22.0 + or - 4.4%, P=0.038). Anti-TLR2 treatment significantly reduced neutrophil, macrophage, and T-lymphocyte infiltration. Furthermore, tumor necrosis factor-alpha, interleukin-1alpha, granulocyte macrophage colony-stimulating factor, and interleukin-10 were significantly reduced, as were phosphorylated c-jun N-terminal kinase, phosphorylated p38 mitogen-activated protein kinase, and caspase 3/7 activity levels.


CONCLUSIONS
Circulating TLR2 expression mediates myocardial ischemia/reperfusion injury. Antagonizing TLR2 just 5 minutes before reperfusion reduces infarct size and preserves cardiac function and geometry. Anti-TLR2 therapy exerts its action by reducing leukocyte influx, cytokine production, and proapoptotic signaling. Hence, monoclonal anti-TLR2 antibody is a potential candidate as an adjunctive for reperfusion therapy in patients with myocardial infarction.
Arslan
Fatih
F
Laboratory of Experimental Cardiology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.
Smeets
Mirjam B
MB
O'Neill
Luke A J
LA
Keogh
Brian
B
McGuirk
Peter
P
Timmers
Leo
L
Tersteeg
Claudia
C
Hoefer
Imo E
IE
Doevendans
Pieter A
PA
Pasterkamp
Gerard
G
de Kleijn
Dominique P V
DP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D017209:Apoptosis / Q000276:immunology; D006410:Hematopoiesis / Q000276:immunology; D007962:Leukocytes / Q000276:immunology* / Q000378:metabolism; D008279:Magnetic Resonance Imaging; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009203:Myocardial Infarction / Q000276:immunology / Q000473:pathology / Q000628:therapy; D015428:Myocardial Reperfusion Injury / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D009205:Myocarditis / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D009206:Myocardium / Q000276:immunology / Q000473:pathology; D015398:Signal Transduction / Q000276:immunology; D013599:Systole; D051195:Toll-Like Receptor 2 / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; C493486:Tlr2 protein, mouse; D051195:Toll-Like Receptor 2