肽抑制人巨细胞病毒感染。
Peptide inhibition of human cytomegalovirus infection.
机构信息
Graduate Program in Biomedical Sciences and Department of Microbiology and Immunology, Tulane University, 1430 Tulane Avenue, New Orleans, LA 70112 USA.
出版信息
Virol J. 2011 Feb 22;8:76. doi: 10.1186/1743-422X-8-76.
BACKGROUND
Human cytomegalovirus (HCMV) is the most prevalent congenital viral infection in the United States and Europe causing significant morbidity and mortality to both mother and child. HCMV is also an opportunistic pathogen in immunocompromised individuals, including human immunodeficiency virus (HIV)- infected patients with AIDS, and solid organ and allogeneic stem cell transplantation recipients. Current treatments for HCMV-associated diseases are insufficient due to the emergence of drug-induced resistance and cytotoxicity, necessitating novel approaches to limit HCMV infection. The aim of this study was to develop therapeutic peptides targeting glycoprotein B (gB), a major glycoprotein of HCMV that is highly conserved across the Herpesviridae family, that specifically inhibit fusion of the viral envelope with the host cell membrane preventing HCMV entry and infection.
RESULTS
Using the Wimley-White Interfacial Hydrophobicity Scale (WWIHS), several regions within gB were identified that display a high potential to interact with lipid bilayers of cell membranes and hydrophobic surfaces within proteins. The ability of synthetic peptides analogous to WWIHS-positive sequences of HCMV gB to inhibit viral infectivity was evaluated. Human foreskin fibroblasts (HFF) were infected with the Towne-GFP strain of HCMV (0.5 MOI), preincubated with peptides at a range of concentrations (78 nm to 100 μM), and GFP-positive cells were visualized 48 hours post-infection by fluorescence microscopy and analyzed quantitatively by flow cytometry. Peptides that inhibited HCMV infection demonstrated different inhibitory concentration curves indicating that each peptide possesses distinct biophysical properties. Peptide 174-200 showed 80% inhibition of viral infection at a concentration of 100 μM, and 51% and 62% inhibition at concentrations of 5 μM and 2.5 μM, respectively. Peptide 233-263 inhibited infection by 97% and 92% at concentrations of 100 μM and 50 μM, respectively, and 60% at a concentration of 2.5 μM. While peptides 264-291 and 297-315, individually failed to inhibit viral infection, when combined, they showed 67% inhibition of HCMV infection at a concentration of 0.125 μM each.
CONCLUSIONS
Peptides designed to target putative fusogenic domains of gB provide a basis for the development of novel therapeutics that prevent HCMV infection.
背景
人类巨细胞病毒（HCMV）是美国和欧洲最常见的先天性病毒感染，对母婴均有显著的发病率和死亡率。HCMV 也是免疫功能低下个体中的机会性病原体，包括感染艾滋病的人类免疫缺陷病毒（HIV）患者，以及实体器官和同种异体干细胞移植受者。由于药物诱导的耐药性和细胞毒性的出现，目前针对 HCMV 相关疾病的治疗方法并不充分，因此需要新的方法来限制 HCMV 感染。本研究旨在开发针对糖蛋白 B（gB）的治疗性肽，gB 是 HCMV 的主要糖蛋白，在疱疹病毒科家族中高度保守，可特异性抑制病毒包膜与宿主细胞膜融合，从而阻止 HCMV 进入和感染。
结果
使用 Wimley-White 界面疏水性标度（WWIHS），鉴定出 gB 中的几个区域具有与细胞膜的脂质双层和蛋白质内部的疏水表面相互作用的高潜力。评估了与 HCMV gB 的 WWIHS 阳性序列类似的合成肽抑制病毒感染力的能力。用 Towne-GFP 株 HCMV（0.5 MOI）感染人包皮成纤维细胞（HFF），用不同浓度（78nm 至 100μM）的肽预先孵育，然后通过荧光显微镜观察感染后 48 小时的 GFP 阳性细胞，并通过流式细胞术进行定量分析。抑制 HCMV 感染的肽显示出不同的抑制浓度曲线，表明每种肽都具有独特的物理化学性质。肽 174-200 在 100μM 浓度下抑制病毒感染 80%，在 5μM 和 2.5μM 浓度下分别抑制 51%和 62%。肽 233-263 在 100μM 和 50μM 浓度下分别抑制感染 97%和 92%，在 2.5μM 浓度下抑制 60%。虽然肽 264-291 和 297-315 单独不能抑制病毒感染，但当联合使用时，它们在 0.125μM 浓度下分别抑制 67%的 HCMV 感染。
结论
设计用于靶向 gB 假定融合结构域的肽为开发预防 HCMV 感染的新型治疗方法提供了基础。
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