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用在恒河猴胎儿肺细胞中制备的纯化、灭活登革2型病毒疫苗原型免疫小鼠后的免疫原性和保护性反应。

Immunogenic and protective response in mice immunized with a purified, inactivated, Dengue-2 virus vaccine prototype made in fetal rhesus lung cells.

作者信息

Putnak R, Cassidy K, Conforti N, Lee R, Sollazzo D, Truong T, Ing E, Dubois D, Sparkuhl J, Gastle W, Hoke C

机构信息

Department of Virus Diseases, Walter Reed Army Institute of Research, Washington, District of Columbia, USA.

出版信息

Am J Trop Med Hyg. 1996 Nov;55(5):504-10. doi: 10.4269/ajtmh.1996.55.504.

Abstract

The feasibility of a purified, inactivated vaccine (PIV) against dengue type 2 (DEN-2) virus was explored. Dengue-2 virus strain 16681 was used for producing a monotypic PIV. Virus adapted to fetal rhesus lung (FRhL-2) cells was harvested from roller bottle culture supernatant fluids, concentrated, and purified on sucrose gradients. Analysis of purified virus preparations by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting showed primarily envelope (E) and premembrane (prM) antigens. These preparations had a purity, estimated from silver-stained gels, of approximately 70%, and a yield, based on recovery of virus and viral antigen, of 10-20%. The purified virus was inactivated with 0.05% formalin at 22 degrees C, or alternatively, with 7 mRads from a 60Co source. Vaccinated mice developed high titers of anti-DEN-2 virus neutralizing antibody and were partially protected from virus challenge. These results warrant further testing and development of PIVs for the other DEN virus serotypes.

摘要

探索了一种针对2型登革热(DEN-2)病毒的纯化灭活疫苗(PIV)的可行性。使用登革热2型病毒株16681来生产单型PIV。从滚瓶培养上清液中收获适应恒河猴胎肺(FRhL-2)细胞的病毒,进行浓缩,并在蔗糖梯度上纯化。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和蛋白质印迹法对纯化的病毒制剂进行分析,结果显示主要为包膜(E)抗原和前膜(prM)抗原。根据银染凝胶估算,这些制剂的纯度约为70%,基于病毒和病毒抗原的回收率,产量为10%-20%。纯化后的病毒在22℃下用0.05%福尔马林灭活,或者用来自60Co源的7毫拉德辐射灭活。接种疫苗的小鼠产生了高滴度的抗DEN-2病毒中和抗体,并受到部分保护,免受病毒攻击。这些结果为进一步测试和开发针对其他登革热病毒血清型的PIV提供了依据。

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