COLD ADAPTATION OF HUMAN ROTAVIRUSES

人类轮状病毒的冷适应

• 批准号: 3803275
• 负责人: Y HOSHINO
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3803275
• 关键词: Rotavirus; cold temperature; genetic strain; rotavirus vaccines; temperature sensitive mutant; tissue /cell culture

Three human rotavirus cell-culture isolates (strains D [VP7:1, VP4:1A], DS-1 [VP7:2, VP4:1B], and 202 [VP7:4, VP4:1B]) as well as two human x human rotavirus reassortants (Wa x P [VP7:3, VP4:1A] and Wa x DS-1 [VP7:2, VP4:1A]) have been successfully adapted to grow in primary African Green monkey kidney cells at the suboptimal temperature of 30 degrees C, 28 degrees C, or 26 degrees C. These rotavirus strains were cold-adapted in order to determine whether cold-adapted (ca) mutants selected by this strategy are attenuated and suitable for use in a live virus vaccine. The D and the Wa x P strains were passaged 10 times either at 28 degrees C or 26 degrees C and are being plaque-purified. Analysis of the efficiency of plaque formation at various temperatures established that the 26 degrees C cold-adapted D strain is indeed a ca mutant; it produced plaques at 26 degrees C, whereas its parent did not. In addition, this mutant is also temperature sensitive having a shut-off temperature for plaque formation of 37 degrees C. The DS-1 strain (passaged 12 times at 28 degrees C) and Wa x DS-1 reassortant (passaged 10 times at 28 degrees C) are also being plaque-purified. Selected human rotaviruses including the D, Wa x P, and Wa x DS-1 have successfully been adapted to growth in DBS-FRhL-2 cells, which is a cell substrate suitable for preparation of vaccines for use in humans.

三种人轮状病毒细胞培养分离株（菌株D [VP 7：1，VP 4：1A]， DS-1 [VP 7：2，VP 4：1B]和202 [VP 7：4，VP 4：1B]）以及两个人x 人轮状病毒抑制剂（Wa × P [VP 7：3，VP 4：1A]和Wa × DS-1 [VP7：2，VP 4：1A]）已成功地适应在初级生长 非洲绿色猴肾细胞在30 ℃的次适温度下 在一些实施方案中，所述温度为20摄氏度、28摄氏度或26摄氏度。这些轮状病毒株是 为了确定冷适应（CA）突变体是否 通过这种策略选择的疫苗是减毒的，适合用于活的 病毒疫苗 将D和Wa × P菌株传代10次 在28摄氏度或26摄氏度下，并且被噬菌斑净化。 不同温度下菌斑形成效率的分析 确定了26摄氏度的冷适应D菌株确实是一个病例， 突变体;它在26摄氏度产生斑块，而它的父母没有。 此外，该突变体也是温度敏感的，具有关闭功能。 37 ℃的蚀斑形成温度。DS-1菌株 （在28 ℃下传代12次）和Wa × DS-1重组子（在28 ℃下传代12次） 10次，28 ℃）也被噬菌斑纯化。 选择 人轮状病毒包括D、Wa x P和Wa x DS-1， 成功地适应DBS-FRhL-2细胞的生长，这是一种细胞 适用于制备人用疫苗的基质。