Pope M, Marsden P A, Cole E, Sloan S, Fung L S, Ning Q, Ding J W, Leibowitz J L, Phillips M J, Levy G A
Departments of Surgery, The University of Toronto, Toronto, Ontario, Canada.
J Virol. 1998 Sep;72(9):7084-90. doi: 10.1128/JVI.72.9.7084-7090.1998.
The strain-specific spectrum of liver disease following murine hepatitis virus type 3 (MHV-3) infection is dependent on inflammatory mediators released by macrophages. Production of nitric oxide (NO) by macrophages has been implicated in resistance to a number of viruses, including ectromelia virus, vaccinia virus, and herpes simplex virus type 1. This study was undertaken to define the role of NO in MHV-3 infection. Gamma interferon-induced production of NO inhibited growth of MHV-3 in a murine macrophage cell line (RAW 264.7). Viral inhibitory activity was reproduced by the NO donor S-nitroso-N-acetyl-DL-penicillamine (SNAP), whereas N-acetyl-DL-pencillamine (NAP), an inactive analog of SNAP, had no effect. Electron microscopy studies confirmed the inhibitory effects of NO on viral replication. Peritoneal macrophages isolated from A/J mice known to be resistant to MHV-3 produced a fivefold-higher level of NO and higher levels of mRNA transcripts of inducible NO synthase in response to gamma interferon than macrophages from susceptible BALB/cJ mice. SNAP inhibited growth of MHV-3 in macrophages from both strains of mice to similar degrees. In vivo inhibition of NO by N-monomethyl-L-arginine resulted in loss of resistance to MHV-3 in A/J mice. These results collectively demonstrate a defect in the production of NO in macrophages from susceptible BALB/cJ mice and define the importance of endogenous NO in resistance to MHV-3 infection in resistant A/J mice.
3型鼠肝炎病毒(MHV-3)感染后肝脏疾病的菌株特异性谱取决于巨噬细胞释放的炎症介质。巨噬细胞产生的一氧化氮(NO)与对多种病毒的抗性有关,包括埃可病毒、痘苗病毒和1型单纯疱疹病毒。本研究旨在确定NO在MHV-3感染中的作用。γ干扰素诱导的NO产生抑制了MHV-3在鼠巨噬细胞系(RAW 264.7)中的生长。NO供体S-亚硝基-N-乙酰-DL-青霉胺(SNAP)重现了病毒抑制活性,而SNAP的无活性类似物N-乙酰-DL-青霉胺(NAP)则没有效果。电子显微镜研究证实了NO对病毒复制的抑制作用。从已知对MHV-3有抗性的A/J小鼠分离的腹腔巨噬细胞,在受到γ干扰素刺激后,产生的NO水平比易感的BALB/cJ小鼠的巨噬细胞高五倍,诱导型NO合酶的mRNA转录水平也更高。SNAP对两种小鼠品系巨噬细胞中MHV-3生长的抑制程度相似。N-单甲基-L-精氨酸在体内对NO的抑制导致A/J小鼠对MHV-3的抗性丧失。这些结果共同证明了易感的BALB/cJ小鼠巨噬细胞中NO产生存在缺陷,并确定了内源性NO在抗性A/J小鼠抵抗MHV-3感染中的重要性。