Intracellular ATP and total adenylate concentrations are critical predictors of reovirus productivity from Vero cells
Intracellular ATP and total adenylate concentrations are critical predictors of reovirus productivity from Vero cells
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DOI：
10.1002/bit.20873
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发表时间：
2006-07-05
影响因子：
3.8
通讯作者：
Butler, M.
中科院分区：
文献类型：
--
作者：
Burgener, A.;Coombs, K.;Butler, M.
在无血清培养基中的Vero细胞培养物中研究了呼肠孤病毒3型Dearing的生产力。病毒生产率取决于细胞的代谢状态，而不是细胞被感染时的生长阶段。通过在营养物耗尽的培养基中孵育24小时来建立处于不同能量状态的细胞。这导致可变的细胞内核苷酸浓度，但保持高细胞活力。在感染时分析的核苷酸中，只有细胞内[ATP]和总腺苷酸核苷酸与病毒生产率呈正相关。相关数据遵循S形曲线，并通过多项式回归分析定义方程。ATP和总腺苷酸的表观阈值分别为3.2 fmol/cell和3.3 fmol/cell，此时病毒产量为最大值的50%。与对照相比，感染时ATP和总腺苷酸水平较低的培养物导致总体病毒滴度降低多达95%。腺苷酸能荷（AEC）与病毒产量呈负相关，AEC值> 0.97，导致病毒产量低。感染点的细胞内ATP或总腺苷酸浓度可用作设计用于病毒/疫苗生产的生物过程中病毒产量的预测因子。(c)2006 Wiley Periodicals，Inc.
The productivity of reovirus type-3 Dearing was studied in cultures of Vero cells in serum-free media. Viral productivity was dependent upon the metabolic state of the cells rather than the phase of growth at which the cells were infected. Cells at different energy states were established by 24-h incubation in nutrient-depleted media. This resulted in variable intracellular nucleotide concentrations but high cellular viability was maintained. Of the nucleotides analyzed at the time of infection only the intracellular [ATP] and total adenylate nucleotides were positively correlated with viral productivity. The correlated data followed a sigmoidal plot with an equation defined by polynomial regression analysis. Apparent threshold values of 3.2 fmol/cell and 3.3 fmol/cell were established for ATP and total adenylate, respectively, at which the viral production was 50% the maximal value. Cultures with lower ATP and total adenylate levels at the time of infection resulted in as much as a 95% reduction in overall viral titer compared to the control. The adenylate energy charge (AEC) showed a negative correlation with viral production with an AEC value > 0.97 resulting in low virus productivity. Intracellular ATP or total adenylate concentration at the point of infection may be used as a predictor of viral yield in bioprocesses designed for virus/vaccine production. (c) 2006 Wiley Periodicals, Inc.