Neutralizing anti-4-1BBL treatment improves cardiac function in viral myocarditis

Abstract

Coxsackievirus B3 (CVB3) is the most common causative agent of infectious myocarditis. Chronic inflammation, loss of contractile tissue, and maladaptive remodeling all contribute to dilated cardiomyopathy and heart failure. The 4-1BB receptor is a costimulatory molecule expressed by T cells and cardiomyocytes. We infected mice with CVB3 to examine if virus infection triggers 4-1BB activation and whether inhibition of this pathway will reduce inflammation and improve heart function. Echocardiography was performed on days 3, 9, 30 and at 10 weeks post-infection (pi) and ejection fraction (EF), left ventricular (LV) wall thickness, contractility, and internal cardiac dimensions were measured. At day 9, reduced rate of wall thickening (30±17 vs 70±19%), increased LV wall thickness (0.15±0.04 vs 0.09±0.01 cm in diastole and 0.19±0.04 vs 0.15±0.02 cm in systole), and reduced cardiac volume (0.013±0.004 vs 0.023±0.003 ml in diastole and 0.004±0.002 ml vs 0.007±0.001 ml in systole) were observed in infected hearts as compared with shams. At 14 days pi, CVB3-infected mice were randomly assigned to receive either anti-4-1BBL neutralizing (M522) or control antibodies (Ab) for 8 weeks. Cardiac damage, fibrosis, and inflammation were assessed by histological stains and immunohistochemistry. Polymerase chain reaction (PCR) was utilized to detect matrix metalloproteinase (MMP)-2, MMP-9, and MMP-12 expressions. At 10 weeks pi, M522 treatment improved LV wall thickening rate (−10±13 vs −49±16%, expressed as percentage change from baseline) and reduced diastolic LV posterior wall thickness (17±10 vs 57±47%, expressed as percentage change from baseline), cardiac damage as assessed by histological scores (0 vs 1.3±1.5), fibrosis by collagen volume fraction (3.2±0.6 vs 4.9±2.2%), overall inflammation (5.9±1.3 vs 8.5±4.1%), and T-cell infiltration (1.3±0.9 vs 4.3±3.8%) as compared to control. MMP-12 was highly increased during acute and chronic myocarditis, but was significantly decreased by M522 treatment. Thus, long-term inhibition of the 4-1BB pathway reduces cardiac damage, remodeling, and inflammation during viral myocarditis.