Dynamics of Biologically Active Subpopulations of Influenza Virus: Plaque-Forming, Noninfectious Cell-Killing, and Defective Interfering Particles
Dynamics of Biologically Active Subpopulations of Influenza Virus: Plaque-Forming, Noninfectious Cell-Killing, and Defective Interfering Particles
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DOI：
10.1128/jvi.02680-08
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发表时间：
2009-08-15
影响因子：
5.4
通讯作者：
Sekellick, Margaret J.
中科院分区：
文献类型：
--
作者：
Marcus, Philip I.;Ngunjiri, John M.;Sekellick, Margaret J.
本文比较了一类新型流感病毒非感染性细胞杀伤颗粒（niCKP）与缺陷干扰颗粒（DIP）在时间形态和数量上的动态变化。在缺乏可检测到的niCKP或DIP的蛋源性MDCK细胞中进行单次高倍数传代后，两类颗粒大量出现（5 × 10(8)/ml），斑块形成颗粒（PFP）滴度下降了60倍。经过两次连续高倍数传代后，DIP保持相对稳定，DIP/niCKP比值达到10:1，PFP下降约1万倍。这些非感染性生物活性颗粒（niBAP）包含在niCKP和DIP亚群中，占总血凝颗粒群的约20%。DIP既不杀死细胞，也不干扰niCKP或PFP（感染性CKP）的细胞杀伤（诱导凋亡）活性，尽管它们阻断了PFP的复制。相对于PFP失活的13600个核苷酸（nt）的紫外线靶标，niCKP的紫外线靶标为2400个核苷酸（nt），与其中一个聚合酶亚基基因一致，DIP的紫外线靶标为350个核苷酸（nt），与DIP介导干扰的小DI-RNA一致。因此，niCKP和DIP被视为不同的颗粒，在感染过程中倾向于形成高多样性。假设这些条件会导致病毒及其包装的临时调节复制发生畸变，从而导致niBAP的产生。DIP与流感病毒的毒力有关，但niCKP的作用尚不清楚。
The dynamic changes in the temporal appearance and quantity of a new class of influenza virus, noninfectious cell-killing particles (niCKP), were compared to defective interfering particles (DIP). After a single high-multiplicity passage in MDCK cells of an egg-derived stock that lacked detectable niCKP or DIP, both classes of particles appeared in large numbers (> 5 x 10(8)/ml), and the plaque-forming particle (PFP) titer dropped similar to 60-fold. After two additional serial high-multiplicity passages the DIP remained relatively constant, the DIP/niCKP ratio reached 10: 1, and the PFP had declined by about 10,000-fold. Together, the niCKP and DIP subpopulations constituted ca. 20% of the total hemagglutinating particle population in which these noninfectious biologically active particles (niBAP) were subsumed. DIP neither killed cells nor interfered with the cell-killing (apoptosis-inducing) activity of niCKP or PFP (infectious CKP), even though they blocked the replication of PFP. Relative to the UV-target of similar to 13,600 nucleotides (nt) for inactivation of PFP, the UV target for niCKP was similar to 2,400 nt, consistent with one of the polymerase subunit genes, and that for DIP was similar to 350 nt, consistent with the small DI-RNA responsible for DIP-mediated interference. Thus, niCKP and DIP are viewed as distinct particles with a propensity to form during infection at high multiplicities. These conditions are postulated to cause aberrations in the temporally regulated replication of virus and its packaging, leading to the production of niBAP. DIP have been implicated in the virulence of influenza virus, but the role of niCKP is yet unknown.