Inhibition of viral replication by nitric oxide and its reversal by ferrous sulfate and tricarboxylic acid cycle metabolites.
Inhibition of viral replication by nitric oxide and its reversal by ferrous sulfate and tricarboxylic acid cycle metabolites.
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DOI：
10.1084/jem.181.6.2171
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发表时间：
1995-06-01
影响因子：
15.3
通讯作者：
Harris, Nicholas
中科院分区：
文献类型：
--
作者：
Karupiah, Gunasegaran;Harris, Nicholas
ifn - γ诱导的小鼠巨噬细胞来源细胞系RAW 264.7中的一氧化氮（NO）先前被证明可以抑制痘病毒、痘苗病毒（VV）和HSV-1的复制。在目前的研究中，我们证明了由于VV感染而激活的小鼠巨噬细胞表达诱导型一氧化氮合酶。这些活化的巨噬细胞对VV感染具有抗性，并有效地阻断了VV和HSV-1在受感染的上皮细胞和成纤维细胞来源的旁观者细胞中的复制。这种抑制作用依赖于精氨酸，与培养物中亚硝酸盐的产生相关，并且可以通过NOS抑制剂N - omega-单甲基- l-精氨酸逆转。用NO供体s-亚硝基-n -乙酰青霉胺处理病毒感染的人293细胞，研究了NO介导的VV复制抑制作用。利用vv特异性DNA探针、针对暂时表达的病毒蛋白的抗体和透射电子显微镜，我们发现NO抑制病毒晚期基因蛋白合成、DNA复制和病毒颗粒形成，但不抑制分析的早期蛋白的表达。通过外源性硫酸亚铁和L-半胱氨酸逆转NO介导的对293细胞VV复制的抑制，确定了NO的可能酶促靶点。抑制的逆转可能源于这些试剂分别保护或再生非血红素铁或巯基的能力，这对于易被NO失活的酶的催化活性是必不可少的。
IFN-gamma-induced nitric oxide (NO) in the murine macrophage-derived cell line RAW 264.7 was previously shown to inhibit replication of the poxviruses ectromelia and vaccinia (VV) and HSV-1. In the current study we demonstrate that murine macrophages activated as a consequence of VV infection express inducible nitric oxide synthase. These activated macrophages were resistant to infection with VV and efficiently blocked the replication of VV and HSV-1 in infected bystander cells of epithelial and fibroblast origin. This inhibition was arginine dependent, correlated with nitrite production in cultures, and reversible by the NOS inhibitor N omega-monomethyl-L-arginine. NO- mediated inhibition of VV replication was studied by treatment of virus- infected human 293 cells with the NO donor S-nitroso-N-acetyl- penicillamine. Using a VV-specific DNA probe, antibodies specific for temporally expressed viral proteins, and transmission electron microscopy, we have shown that NO inhibited viral late gene protein synthesis, DNA replication, and virus particle formation, but not expression of the early proteins that were analyzed. Putative enzymatic targets of NO were identified by reversing the NO-mediated inhibition of VV replication in the 293 cells with exogenous ferrous sulfate and L- cysteine. Reversal of inhibition may derive from the capacity of these reagents to protect or regenerate nonheme iron or thiol groups, respectively, which are essential for the catalytic activities of enzymes susceptible to inactivation by NO.
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影响因子：
64.5
作者：
LOWENSTEIN, CJ;SNYDER, SH
通讯作者：
SNYDER, SH
影响因子：
5.4
作者：
KARUPIAH, G;FREDRICKSON, TN;BULLER, RML
通讯作者：
BULLER, RML
影响因子：
56.9
作者：
KARUPIAH, G;XIE, QW;MACMICKING, JD
通讯作者：
MACMICKING, JD
影响因子：
5.4
作者：
SODEIK, B;GRIFFITHS, G;DOMS, RW
通讯作者：
DOMS, RW
DOI：
10.1084/jem.174.4.761
发表时间：
1991-10-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kwon NS;Stuehr DJ;Nathan CF
通讯作者：
Nathan CF