Nolan Sheila M, Surman Sonja R, Amaro-Carambot Emerito, Collins Peter L, Murphy Brian R, Skiadopoulos Mario H
Laboratory of Infectious Diseases, Respiratory Viruses Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 50 South Drive, Building 50, Room 6509, MSC 8007, Bethesda, MD 20892, USA.
Vaccine. 2005 Sep 15;23(39):4765-74. doi: 10.1016/j.vaccine.2005.04.043.
Live-attenuated recombinant human parainfluenza virus type 2 (rHPIV2) vaccine candidates were created using reverse genetics by importing known attenuating mutations in the L polymerase protein from heterologous paramyxoviruses into the homologous sites of the HPIV2 L protein. Four recombinants (rF460L, rY948H, rL1566I, and rS1724I) were recovered and three were attenuated for replication in hamsters. The genetic stability of the imported mutations at three of the four sites was enhanced by use of alternative codons or by deletion of a pair of amino acids. rHPIV2s bearing these modified mutations exhibited enhanced attenuation. The genetically stabilized mutations conferring a high level of attenuation will be useful in generating a live-attenuated virus vaccine for HPIV2.
通过反向遗传学技术，将来自异源副粘病毒的L聚合酶蛋白中已知的减毒突变导入人副流感病毒2型（HPIV2）L蛋白的同源位点，构建了减毒活重组人副流感病毒2型（rHPIV2）候选疫苗。回收了四种重组体（rF460L、rY948H、rL1566I和rS1724I），其中三种在仓鼠体内复制时减毒。通过使用替代密码子或删除一对氨基酸，增强了四个位点中三个位点导入突变的遗传稳定性。携带这些修饰突变的rHPIV2表现出更强的减毒效果。赋予高水平减毒的遗传稳定突变将有助于生产HPIV2减毒活病毒疫苗。
