Putnak R, Cassidy K, Conforti N, Lee R, Sollazzo D, Truong T, Ing E, Dubois D, Sparkuhl J, Gastle W, Hoke C
Department of Virus Diseases, Walter Reed Army Institute of Research, Washington, District of Columbia, USA.
Am J Trop Med Hyg. 1996 Nov;55(5):504-10. doi: 10.4269/ajtmh.1996.55.504.
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.
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。
