Preconditioning by toll-like receptor 2 agonist Pam3CSK4 reduces CXCL1-dependent leukocyte recruitment in murine myocardial ischemia/reperfusion injury.

Objective: To test whether preconditioning with a toll-like receptor (TLR) 2 agonist protects against myocardial ischemia and reperfusion by interfering with chemokine CXCL1 release from cardiomyocytes. Design: C3H mice were challenged with vehicle or synthetic TLR2 agonist Pam(3)Cys-Ser-Lys(4) (Pam(3)CSK(4); 1 mg/kg) 24 hrs before myocardial ischemia (20 mins) and reperfusion (2 hrs or 24 hrs). Infarct size, troponin T release, and leukocyte recruitment were quantified. In murine cardiomyocytes (HL-1), we studied the expression/activation profile of TLR2 in response to stimulation with Pam(3)CSK(4) (0.01-1 mg/mL). Furthermore, we studied the chemokine ligand 1 (CXCL1) response to Pam(3)CSK(4) and ischemia/reperfusion in vivo and in vitro. Setting: University hospital research laboratory. Subjects: Anesthetized male mice and murine cardiomyocytes. Measurements and Main Results: Preconditioning by Pam(3)CSK(4) reduced infarct size and troponin T release. This was accompanied by a decreased recruitment of leukocytes into the ischemic area and an improved cardiac function. In HL-1 cells, TLR2 activation amplified the expression of the receptor in a time-dependent manner and led to CXCL1 release in a concentration-dependent manner. Preconditioning by Pam(3)CSK(4) impaired CXCL1 release in response to a second inflammatory stimulus in vivo and in vitro. Conclusions: Preconditioning by TLR2 agonist Pam(3)CSK(4) reduces myocardial infarct size after myocardial ischemia/reperfusion. One of the mechanisms involved is a diminished chemokine release from cardiomyocytes, which subsequently limits leukocyte infiltration. (Crit Care Med 2010; 38: 903-909)