Viral myocarditis (VMC) is an inflammation of heart muscle in infants and young adolescents. This study explored the function of halofuginone (HF) in Coxsackievirus B3 (CVB3)-treated suckling mice. HF-treated animal exhibited higher survival rate, lower heart/body weight, and more decreased blood sugar concentration than CVB3 group. HF also reduced the expressions of interleukin(IL)-17 and IL-23 and the numbers of Th17 cells. Moreover, HF downregulated pro-inflammatory cytokine levels and increased anti-inflammatory cytokine levels. The expressions of transforming growth factor(TGF-beta 1) and nuclear factor kappa-light-chain-enhancer of activated B (NF-kappa B) p65/tumor necrosis factor-alpha (TNF-alpha) proteins were decreased by HF as well. Finally, the overexpression of TGF-beta 1 counteracted the protection effect of HF in CVB3-treated suckling mice. In summary, our study suggests HF increases the survival of CVB3 suckling mice, reduces the Th17 cells and pro-inflammatory cytokine levels, and may through downregulation of the TGF-beta 1-mediated expression of NF-kappa B p65/TNF-alpha pathway proteins. These results offer a potential therapeutic strategy for the treatment of VMC. (C) 2016 Elsevier Inc. All rights reserved.