Antibodies against synthetic peptides of herpes simplex virus type 1 glycoprotein D and their capability to neutralize viral infectivity in vitro
Antibodies against synthetic peptides of herpes simplex virus type 1 glycoprotein D and their capability to neutralize viral infectivity in vitro
复制标题
1 型单纯疱疹病毒糖蛋白 D 合成肽的抗体及其体外中和病毒感染性的能力
作者：
J. Drijfhout;H. Geerligs;Wim Bloemhoff;Matty;Feijlbrief;A. Cees;Bos;Peter Hoogerhout;Karel E. T. Kerling;T. Popken;Kunja Slopsema;Jan B. Wilterdink;W. Welling
对应于 1 型单纯疱疹病毒 (HSV-1) 糖蛋白 D 残基 1-13、9-21、18-30、82-93、137-150、181-197、232-243、235-243、267-281、271-281 和 302-315 的肽为化学合成的。这些肽与载体蛋白偶联，所得缀合物用于免疫兔子。使用酶联免疫吸附测定法测定免疫后收集的血清中的抗肽抗体滴度。所有肽似乎对兔子都具有免疫原性。使用 HSV-1 感染的 Vero 细胞去污剂提取物进行的蛋白质免疫印迹分析表明，针对每种肽的抗体能够在变性条件下与亲本糖蛋白发生反应。针对肽1-13、9-21和18-30的抗血清在体外中和了HSV-1感染性，肽9-21在这方面是最成功的。用肽9-21和267-281的混合物进行免疫产生了抗血清，该抗血清在蛋白质印迹分析中与糖蛋白gD强烈反应，并在体外显示出更牢固的病毒中和活性。
Peptides corresponding to residues 1-13, 9-21, 18-30, 82-93, 137-150, 181-197, 232-243, 235-243, 267-281, 271-281 and 302-315 of glycoprotein D of herpes simplex virus type 1 (HSV-1) were chemically synthesized. These peptides were coupled to carrier proteins, and the resulting conjugates were used to immunize rabbits. An enzyme-linked immunosorbent assay was used to determine antipeptide antibody titers in serum collected after immunization. All peptides appeared to be immunogenic in rabbits. Western immunoblot analysis with detergent extracts of HSV-1-infected Vero cells showed that antibodies against each of the peptides were able to react with the parent glycoprotein under denaturing conditions. Antisera against peptides 1-13, 9-21, and 18-30 neutralized HSV-1 infectivity in vitro, peptide 9-21 being the most successful in this respect. Immunization with a mixture of peptides 9-21 and 267-281 yielded antisera which reacted strongly with glycoprotein gD in Western blot analysis and showed a more solid virus-neutralizing activity in vitro.