Ma Jia, Kennedy-Stoskopf Suzanne, Jaynes Jesse M, Thurmond Linda M, Tompkins Wayne A
Department of Microbiology, Pathology and Parasitology, North Carolina State University, Raleigh, North Carolina, USA.
J Virol. 2002 Oct;76(19):9952-61. doi: 10.1128/jvi.76.19.9952-9961.2002.
Natural peptide antibiotics are part of host innate immunity against a wide range of microbes, including some viruses. Synthetic peptides modeled after natural peptide antibiotics interfere with microbial membranes and are termed peptidyl membrane-interactive molecules (peptidyl-MIM [Demegen Inc, Pittsburgh, Pa.]). Sixteen peptidyl-MIM candidates were tested for activity against feline immunodeficiency virus (FIV) on infected CrFK cells. Three of them (D4E1, DC1, and D1D6) showed potent anti-FIV activity in chronically infected CrFK cells as measured by decreased reverse transcriptase (RT) activity, having 50% inhibitory concentrations of 0.46, 0.75, and 0.94 micro M, respectively, which were approximately 10 times lower than their direct cytotoxic concentrations. Treatment of chronically infected CrFK cells with 2 micro M D4E1 for 3 days completely reversed virus-induced cytopathic effect. Immunofluorescence revealed reduced p26 staining in these cells. Treatment of chronically infected CrFK cells with 2 micro M D4E1 suppressed virus production ( approximately 50%) for up to 7 days, The virions from the D4E1-treated culture had impaired infectivity, as measured by the 50% tissue culture infectious dose and nested PCR analysis of proviral DNA. However, these noninfectious virions were able to bind and internalize, suggesting a defect at some postentry step. After chronically infected CrFK cells were treated with D4E1 for 24 h, increased cell-associated mature p26 Gag and decreased extracellular virus-associated p26 Gag were observed by Western blot analysis, suggesting that virus assembly and/or release may be blocked by D4E1 treatment, whereas virus binding, penetration, RNA synthesis, and protein synthesis appear to be unaffected. Synthetic peptide antibiotics may be useful tools in the search for antiviral drugs having a wide therapeutic window for host cells.
天然肽抗生素是宿主针对包括某些病毒在内的多种微生物的固有免疫的一部分。模仿天然肽抗生素构建的合成肽会干扰微生物膜,被称为肽基膜相互作用分子(肽基-MIM [德梅根公司,匹兹堡,宾夕法尼亚州])。对16种肽基-MIM候选物进行了测试,以检测其在感染猫免疫缺陷病毒(FIV)的CrFK细胞上的活性。其中三种(D4E1、DC1和D1D6)在慢性感染的CrFK细胞中表现出强大的抗FIV活性,通过逆转录酶(RT)活性降低来衡量,其50%抑制浓度分别为0.46、0.75和0.94微摩尔,比其直接细胞毒性浓度低约10倍。用2微摩尔D4E1处理慢性感染的CrFK细胞3天可完全逆转病毒诱导的细胞病变效应。免疫荧光显示这些细胞中p26染色减少。用2微摩尔D4E1处理慢性感染的CrFK细胞可抑制病毒产生(约50%)长达7天。通过50%组织培养感染剂量和原病毒DNA的巢式PCR分析测定,来自D4E1处理培养物的病毒粒子感染性受损。然而,这些无感染性的病毒粒子能够结合并内化,表明在进入后的某个步骤存在缺陷。慢性感染的CrFK细胞用D4E1处理24小时后,通过蛋白质印迹分析观察到细胞相关的成熟p26 Gag增加,细胞外病毒相关的p26 Gag减少,这表明D4E1处理可能会阻断病毒组装和/或释放,而病毒结合、穿透、RNA合成和蛋白质合成似乎未受影响。合成肽抗生素可能是寻找对宿主细胞具有广泛治疗窗的抗病毒药物的有用工具。